interacts_with DB01169?
Metallothionein prevention of arsenic trioxide-induced cardiac cell death is associated with its inhibition of mitogen-activated protein kinases activation in vitro and in vivo . Cardiotoxicity induced by arsenic trioxide has become a serious blockade of clinical applications of this effective anticancer agent . The general mechanism responsible for arsenic cardiotoxicity has been attributed to its induction of oxidative stress . Metallothionein ( MT ) has been extensively proven to be a potent endogenous antioxidant that protects heart against oxidative stress-induced cardiac damage . To investigate whether and how MT protects against arsenic cardiotoxicity , MT-overexpressing H9c2 ( MT-H9c2 ) cardiac cells and transgenic ( MT-TG ) mice with their corresponding controls were exposed to the clinical relevant dose of arsenic trioxide . Cardiac cell apoptosis was detected by molecular indices , including the cleavage of caspase 3 and caspase 12 , Bax/Bcl2 expression ratio , P35638 expression and/or confirmed by a terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay . DB01169 dose- and time-dependently induced cardiac cell death in H9c2 cells with a significant activation of major MAPK subfamily members such as P27361 /2 , JNK and p38 , but not in MT-H9c2 cells . Importantly , the protective effect of MT on arsenic trioxide-induced apoptotic cell death was completely recaptured in the heart of MT-TG with a significant prevention of MAPKs activation . These results indicate that arsenic trioxide-upregulated MAPKs might play important role in arsenic trioxide-induced apoptotic cell death in cardiac cells both in vivo and in vitro , and MT 's suppression of arsenic trioxide apoptotic effect was associated with the inhibition of MAPK activation . Therefore , selective elevation of cardiac MT levels with pharmacological approaches may be a potential strategy for the prevention of arsenic cardiotoxicity . Dissection of the phenotypic and genotypic associations with nicotinic dependence . INTRODUCTION : Strong evidence demonstrates that nicotine dependence is associated with 4 genetic variants rs16969968 , rs6474412 , rs3733829 , and rs1329650 in large-scale Genome-Wide Association Studies . We examined how these identified genetic variants relate to nicotine dependence defined by different categorical and dimensional measures . METHODS : Four genetic variants were analyzed in 2,047 subjects of European descent ( 1,062 cases and 985 controls ) . DB00184 dependence was assessed with multiple smoking measures , including the Fagerström Test for DB00184 Dependence , the Diagnostic and Statistical Manual for Mental Disorders-IV ( DSM-IV ) nicotine dependence , the DB00184 Dependence Syndrome Scale , and the Wisconsin Inventory of Smoking Dependence Motives . Single-item measures of cigarettes per day ( O75976 ) and time to first cigarette ( Q15669 ) in the morning were also examined . RESULTS : Among the variants , association effect sizes were largest for rs16969968 , with measures of craving and heavy smoking , especially cigarettes smoked per day , showing the largest effects . Significant but weaker associations were found for rs6474412 and rs3733729 but not for rs1329650 . None of the more comprehensive measures of smoking behaviors yielded stronger genetic associations with these variants than did O75976 . CONCLUSIONS : O75976 is an important simple measure that captures in part the genetic associations of P30532 and nicotine dependence , even when other more comprehensive measures of smoking behaviors are examined . The P30532 gene is associated with heavy compulsive smoking and craving ; this should inform the mission to improve the diagnostic validity of DSM-V . Combination gene therapy targeting on interleukin-1β and O14788 for wear debris-induced aseptic loosening . This study investigated the efficacy of a combination gene therapy to repress interleukin-1 ( IL-1 ) and receptor activator of nuclear factor NF-kappa B ligand ( O14788 ) for the treatment of particulate debris-induced aseptic loosening , and tried to explore the molecular mechanism of the exogenous gene modifications on osteoclastogenesis . RAW cells activated by titanium particles were transduced with DFG-IL-1Ra ( retroviral vector encoding IL-1 receptor antagonist ) and AAV- O00300 ( adeno-associated viral vectors-osteoprotegerin ) individually or in combination for 4 weeks . Pro-inflammatory cytokines in culture media were determined by enzyme-linked immunosorbent assay , and gene expressions of Q9Y6Q6 , IL-1β , c-Fos , Q9Y4K3 , P45983 and CPK were examined using real-time PCR . An established knee-implant-failure mouse model was employed to evaluate the efficacy of the in vivo double-gene therapy . The surgical implantation of a titanium alloy pin into the proximal tibia was followed by monthly challenge with titanium debris . Peri-implant gene transfers of IL-1Ra and O00300 ( respectively or in combination ) were given 3 weeks after surgery . The combination of O00300 and IL-1Ra gene transfer exhibited strong synergetic effects in blockage of inflammation and osteoclastogenesis at 8 weeks after gene modification . The combination therapy reversed peri-implant bone resorption and restored implant stability when compared with either single gene transduction . Real-time PCR data indicated that the action of IL-1Ra gene therapy may be mediated via the P45983 pathway , while the reduction of osteoclastogenesis by O00300 gene modification may be regulated by c-Fos expression . In addition , both gene modifications resulted in significant diminishment of Q9Y4K3 expression . TG-interacting factor transcriptionally induced by AKT/ O43524 is a negative regulator that antagonizes arsenic trioxide-induced cancer cell apoptosis . DB01169 ( ATO ) is a multi-target drug approved by the Food and Drug Administration as the first-line chemotherapeutic agent for the treatment of acute promyelocytic leukemia . In addition , several clinical trials are being conducted with arsenic-based drugs for the treatment of other hematological malignancies and solid tumors . However , ATO 's modest clinical efficacy on some cancers , and potential toxic effects on humans have been reported . Determining how best to reduce these adverse effects while increasing its therapeutic efficacy is obviously a critical issue . Previously , we demonstrated that the JNK-induced complex formation of phosphorylated c-Jun and TG-interacting factor ( Q15583 ) antagonizes P29323 -induced cyclin-dependent kinase inhibitor P38936 ( P38936 ( P38936 /CIP1) ) expression and resultant apoptosis in response to ATO in A431 cells . Surprisingly , at low-concentrations ( 0.1-0.2 μM ) , ATO increased cellular proliferation , migration and invasion , involving Q15583 expression , however , at high-concentrations ( 5-20 μM ) , ATO induced cell apoptosis . Using a promoter analysis , Q15583 was transcriptionally regulated by ATO at the O43524 binding site ( -1486 to -1479bp ) via the c-Src/ P00533 /AKT pathway . Stable overexpression of Q15583 promoted advancing the cell cycle into the S phase , and attenuated 20 μM ATO-induced apoptosis . Furthermore , blockage of the AKT pathway enhanced ATO-induced P38936 expression and resultant apoptosis in cancer cells , but overexpression of P31749 inhibited P38936 expression . Therefore , we suggest that Q15583 is transcriptionally regulated by the c-Src/ P00533 /AKT pathway , which plays a role as a negative regulator in antagonizing ATO-induced P38936 expression and resultant apoptosis . Suppression of these antagonistic effects might be a promising therapeutic strategy toward improving clinical efficacy of ATO . DB01169 inhibits osteosarcoma cell invasiveness via MAPK signaling pathway . DB01169 ( As(2)O(3) ) is an active ingredient in traditional Chinese medicine . Recent studies showed that it causes apoptosis in several cancer cells . However , research of As(2)O(3) in osteosarcoma is sparse . In our present study , an inhibitory effect of As(2)O(3) on osteosarcoma cell adhesion and metastasis was observed with a cell adhesion , migration and invasion test . The impact of As(2)O(3) on the activities of P14780 and MAPK pathway-related downstream factors was analyzed by western blotting . Our results showed that As(2)O(3) significantly inhibited motility , migration and invasion in Q9UKB1 and MNNG cells in a concentration-dependent manner at concentrations ranging from 0.5-2 μM , and led to cytoskeletal rearrangements . As(2)O(3) exerted an inhibitory effect on the phosphorylation of P27361 /2 and MEK , which are the members of the MAPK family . Additionally , treatment with As(2)O(3) in combination with inhibitors specific for MEK ( U0126 ) in Q9UKB1 and MNNG cells resulted in a marked inhibition of cell invasion and As(2)O(3) could significantly reduce PMA-induced invasion . In conclusion , we demonstrate the inhibitory effects of As(2)O(3) on the invasiveness of Q9UKB1 and MNNG cells , which may be due at least partly to inactivation of the MAPK signaling pathway . P02649 : a potent inhibitor of endothelial and tumor cell proliferation . Recombinant human apolipoprotein E3 ( apoE ) , purified from E. coli , inhibited the proliferation of several cell types , including endothelial cells and tumor cells in a dose- and time-dependent manner . ApoE inhibited both de novo DNA synthesis and proliferation as assessed by an increase in cell number . Maximal inhibition of cell growth by apoE was achieved under conditions where proliferation was dependent on heparin-binding growth factors . Thus , at low serum concentrations ( 0-2.5 % ) basic fibroblast growth factor ( P09038 ) stimulated the proliferation of bovine aortic endothelial ( BAE ) cells severalfold . The P09038 -dependent proliferation was dramatically inhibited by apoE with an IC50 approximately 50 nM . Under conditions where cell proliferation was mainly serum-dependent , apoE also suppressed growth but required higher concentrations to be effective ( IC50 approximately 500 nM ) . ApoE also inhibited growth of bovine corneal endothelial cells , human melanoma cells , and human breast carcinoma cells . The IC50 values obtained with these cells were generally 3-5 times higher than with BAE cells . Inhibition of cell proliferation by apoE was reversible and dependent on the time of apoE addition to the culture . In addition , apoE inhibited the chemotactic response of endothelial cells that were induced to migrate by a gradient of soluble P09038 . Inhibition of cell proliferation by apoE may be mediated both by competition for growth factor binding to proteoglycans and by an antiadhesive activity of apoE . The present results demonstrate that apoE is a potent inhibitor of proliferation of several cell types and suggest that apoE may be effective in modulating angiogenesis , tumor cell growth , and metastasis . DB01169 ( ATO ) and Q02750 inhibition synergize to induce apoptosis in acute promyelocytic leukemia cells . Recent studies suggest that components of the prosurvival signal transduction pathways involving the Ras-mitogen-activated protein kinase ( MAPK ) can confer an aggressive , apoptosis-resistant phenotype to leukemia cells . In this study , we report that acute promyelocytic leukemia ( APL ) cells exploit the Ras-MAPK activation pathway to phosphorylate at Ser112 and to inactivate the proapoptotic protein Bad , delaying arsenic trioxide ( ATO ) -induced apoptosis . Both in APL cell line NB4 and in APL primary blasts , the inhibition of extracellular signal-regulated kinases 1/2 ( P27361 /2 ) and Bad phosphorylation by Q02750 inhibitors enhanced apoptosis in ATO-treated cells . We isolated an arsenic-resistant NB4 subline ( NB4-As(R) ) , which showed stronger P27361 /2 activity ( 2.7-fold increase ) and Bad phosphorylation ( 2.4-fold increase ) compared to parental NB4 cells in response to ATO treatment . Upon ATO exposure , both NB4 and NB4-As(R) cell lines doubled protein levels of the death antagonist Bcl-xL , but the amount of free Bcl-xL that did not heterodimerize with Bad was 1.8-fold greater in NB4-As(R) than in the parental line . Q02750 inhibitors dephosphorylated Bad and inhibited the ATO-induced increase of Bcl-xL , overcoming ATO resistance in NB4-As(R) . These results may provide a rationale to develop combined or sequential Q02750 inhibitors plus ATO therapy in this clinical setting . DB06643 for joints and bones . DB06643 is an investigational , fully human monoclonal antibody with a high affinity and specificity for receptor activator of nuclear factor kappaB ligand ( O14788 ) , a cytokine member of the tumor necrosis factor family . O14788 , an essential mediator of osteoclast formation , function , and survival , plays a major role in the pathogenesis of postmenopausal osteoporosis , structural damage in rheumatoid arthritis , and bone loss associated with other skeletal disorders . DB06643 suppresses bone turnover by inhibiting the action of O14788 on osteoclasts . DB06643 reduces bone turnover and increases bone mineral density in postmenopausal women with low bone mineral density , reduces fracture risk in women with postmenopausal osteoporosis , and inhibits structural damage in patients with rheumatoid arthritis when added to ongoing methotrexate treatment . It is generally well tolerated , with a good safety profile . Adverse and serious adverse events , including infections and malignancy , are similar in patients treated with denosumab or placebo . Extracellular signal-regulated kinase and the small GTP-binding protein , Rac , contribute to the effects of transforming growth factor-beta1 on gene expression . The kinases and regulatory proteins that convey signals initiated by transforming growth factor-beta ( TGF-beta ) to the nucleus are poorly characterized . To study the role of the extracellular signal-regulated kinase ( P29323 ) pathway in this process , we transiently transfected NIH 3T3 fibroblasts with TGF-beta-responsive luciferase reporter genes and expression vectors designed to interrupt this kinase cascade . Mitogen-activated protein ( Q96HU1 ) kinase phosphatase-1 and a dominant negative Q96HU1 / P29323 kinase 1 mutant reduced stimulation of plasminogen activator inhibitor-1 ( P05121 ) promoter activity by TGF-beta1 from 11.5- to 4-fold and 4.9-fold , respectively . Similar results were observed with the type I collagen promoters . TGF-beta1 increased P27361 activity 4.5-fold at 5 min and 3 . 1-fold at 3 h , while Jun kinase and p38 activity were not affected . Cotransfection of a dominant negative mutant of the small G protein , Rac , but not dominant negative Ras , Cdc42 , or Rho mutants , reduced the effects of TGF-beta1 on the P05121 promoter by approximately half . In support of a role for Rac in signaling by TGF-beta , GTP binding to Rac was increased 3.7-fold following exposure of NIH 3T3 cells to TGF-beta1 for 3 min . These findings indicate that TGF-beta1 modulates gene expression partly through P29323 and Rac in NIH 3T3 cells . Activation of the p38 MAPK/Akt/ P27361 /2 signal pathways is required for the protein stabilization of Q99741 and cyclin D1 in low-dose arsenite-induced cell proliferation . DB01169 ( ATO ) is a first-line anti-cancer agent for acute promyelocytic leukemia , and induces apoptosis in other solid cancer cell lines including breast cancer cells . However , as with arsenites found in drinking water and used as raw materials for wood preservatives , insecticides , and herbicides , low doses of ATO can induce carcinogenesis after long-term exposure . At 24 h after exposure , ATO ( 0.01-1 µM ) significantly increased cell proliferation and promoted cell cycle progression from the P55008 to S/G2 phases in the non-tumorigenic MCF10A breast epithelial cell line . The expression of 14 out of 96 cell-cycle-associated genes significantly increased , and seven of these genes including cell division cycle 6 ( Q99741 ) and cyclin D1 ( P24385 ) were closely related to cell cycle progression from P55008 to S phase . Low-dose ATO steadily increased gene transcript and protein levels of both Q99741 and cyclin D1 in a dose- and time-dependent manner . Low-dose ATO produced reactive oxygen species ( ROS ) , and activated the p38 MAPK , Akt , and P27361 /2 pathways at different time points within 60 min . Small molecular inhibitors and siRNAs inhibiting the activation of p38 MAPK , Akt , and P27361 /2 decreased the ATO-increased expression of Q99741 protein . Inhibiting the activation of Akt and P27361 /2 , but not p38 MAPK , decreased the ATO-induced expression of cyclin D1 protein . This study reports for the first time that p38 MAPK/Akt/ P27361 /2 activation is required for the protein stabilization of Q99741 in addition to cyclin D1 in ATO-induced cell proliferation and cell cycle modulation from P55008 to S phase . Inhibition of P29323 activation enhances the repair of double-stranded breaks via non-homologous end joining by increasing P78527 activation . Non-homologous end joining ( NHEJ ) is one of the major pathways that repairs double-stranded DNA breaks ( DSBs ) . Activation of DNA-PK is required for NHEJ . However , the mechanism leading to P78527 activation remains incompletely understood . We provide evidence here that the MEK- P29323 pathway plays a role in P78527 -mediated NHEJ . In comparison to the vehicle control ( DB01093 ) , etoposide ( ETOP ) -induced DSBs in MCF7 cells were more rapidly repaired in the presence of U0126 , a specific MEK inhibitor , based on the reduction of γ P16104 and tail moments . Additionally , U0126 increased reactivation of luciferase activity , which resulted from the repair of restriction enzyme-cleaved DSBs . Furthermore , while inhibition of P29323 activation using the dominant-negative MEK1K97M accelerated the repair of DSBs , enforcing P29323 activation with the constitutively active MEK1Q56P reduced DSB repair . In line with MEK activating P27361 and P28482 kinases , knockdown of either P27361 or P28482 increased DSB repair . Consistent with the activation of P78527 being required for NHEJ , we demonstrated that inhibition of P29323 activation using U0126 , MEK1K97M , and knockdown of P27361 or P28482 enhanced ETOP-induced activation of P78527 . Conversely , enforcing P29323 activation by MEK1Q56P reduced ETOP-initiated P78527 activation . Taken together , we demonstrate that P29323 reduces NHEJ-mediated repair of DSBs via attenuation of P78527 activation . Kirenol stimulates osteoblast differentiation through activation of the BMP and Wnt/β-catenin signaling pathways in MC3T3-E1 cells . Kirenol has been reported to possess anti-oxidant , anti-inflammatory , anti-allergic , anti-adipogenic , and anti-arthritic activities ; however , its effect on osteoblast differentiation has not yet been reported . The aim of the present study was to evaluate the effect of kirenol on osteoblast differentiation through activation of the bone morphogenetic protein ( BMP ) and Wnt/β-catenin signaling pathways in MC3T3-E1 cells . Kirenol markedly promoted alkaline phosphatase ( ALP ) activity and mineralization . Kirenol not only increased the expression of osteoblast differentiation markers , such as ALP , type I collagen ( ColA1 ) , and osteopontin ( P10451 ) , but also increased the expression of osteoprotegerin/receptor activator of nuclear factor kappa B ligand ( O00300 / O14788 ) ratio . The effects of kirenol on osteoblast differentiation were accompanied by stimulating the expression of the BMP and Wnt/β-catenin signaling pathways , including P12643 , runt-related transcription factor 2 ( Runx2 ) , osterix ( Osx ) , low density lipoprotein receptor related protein 5 ( O75197 ) , disheveled 2 ( O14641 ) , β-catenin , cyclin D1 ( P24385 ) , and phosphorylated glycogen synthase kinase 3β ( GSK3β ) . In addition , kirenol up-regulated the expression of β-catenin , P24385 , ALP , and ColA1 which were down-regulated by siRNA knockdown of β-catenin . Overall , these results demonstrate that kirenol is capable of promoting osteoblast differentiation in MC3T3-E1 cells through activation of the BMP and Wnt/β-catenin signaling pathways , suggesting that it is a potential candidate target for treating or preventing osteoporosis . P14921 , NFkappaB and P05412 synergistically transactivate the human GM- P04141 promoter . Activation of helper T cells results in coordinate expression of a number of cytokines involved in differentiation , proliferation and activation of the haematopoietic system . Granulocyte-macrophage colony stimulating factor ( GM- P04141 ) is one such cytokine , whose increased expression results mostly from increases in transcription . Cis-acting elements with NFkappaB , P05412 and ETS-like binding motifs have been identified in the promoter region of the GM- P04141 gene , and are important or essential for transcriptional activity following T cell activation . P14921 is a transcription factor of the ETS family that is expressed in T cells . We have previously shown that P14921 can transactivate GM- P04141 in Jurkat T cells , but only after the cells have been stimulated by treatment with PMA and ionomycin , agents that mimic T cell activation . Thus we proposed that P14921 , which is expressed constitutively in Jurkat cells , may act in concert with PMA/ionomycin inducible factors . Here we show that P14921 can transactivate a GM- P04141 reporter construct in unstimulated Jurkat cells , providing that either NFkappaB or P05412 transcription factors are supplied by co-transfection . We confirm that binding of endogenous NFkappaB and P05412 is induced following PMA/ionomycin treatment of T cells . Transactivation by P14921 , NFkappaB and P05412 is synergistic , and mutation of the individual binding sites reveals that the transcriptional activities of these factors are interdependent . Our results suggest that constitutive P14921 , and inducible NFkappaB and P05412 , cooperate as part of a higher order transcriptional complex in activated T cells . O75915 is required for arsenic trioxide induced apoptosis in HeLa and MCF-7 cells via reactive oxygen species and mitochondria linked signal pathway . DB01169 , emerging as a standard therapy for refractory acute promyelocytic leukemia , induces apoptosis in a variety of malignant cell lines . O75915 , a novel retinoic acid-inducible gene , is known to be involved in apoptosis induced by various agents , for example , 12-O-tetradecanoylphorbol 13-acetate , N-4-hydroxy-phenyl-retinamide and arsenic trioxide . However , the molecular mechanisms underlying how O75915 gene is functionally involved in apoptosis remain largely unknown . Herein , our studies demonstrated that treatment of arsenic trioxide produced apoptosis in HeLa and MCF-7 cells in a dose-dependent manner and paralleled with increased O75915 expression . O75915 expression was dependent upon generation of intracellular reactive oxygen species induced by arsenic trioxide . Knockdown of O75915 attenuated arsenic trioxide induced apoptosis , and was accompanied by significantly reduced activity of caspase-9 , enhanced Bad phosphorylation and inhibited Q02750 /2 , P27361 /2 and JNK phosphorylations . DB01169 induced loss of mitochondrial transmembrane potential was O75915 -dependent . These findings suggest that O75915 may serve as a pro-apoptotic molecule to mediate arsenic trioxide triggered apoptosis via a reactive oxygen species and mitochondria-associated signal pathway . DB01169 concentration determines the fate of Ewing 's sarcoma family tumors and neuroblastoma cells in vitro . DB01169 ( As(2)O(3) ) induces both the differentiation and apoptosis of acute promyelocytic leukemia cells in a concentration dependent manner . We assessed the effects of As(2)O(3) in CADO-ES Ewing 's sarcoma ( ES ) , JK-GMS peripheral primitive neuroectodermal tumor ( PNET ) , and SH-SY5Y neuroblastoma cells , as they share common histogenetic backgrounds . As(2)O(3) at low concentrations ( 0.1-1 microM ) induced SH-SY5Y differentiation , and whereas PNET cells acquired a slightly differentiated phenotype , change was minimal in ES cells . P28482 ( P28482 ) was activated at low As(2)O(3) concentrations , and PD98059 , an inhibitor of MEK-1 , blocked SH-SY5Y cell differentiation by As(2)O(3) . High concentrations ( 2-10 microM ) of As(2)O(3) induced the apoptosis in all three cell lines , and this was accompanied by the activation of c-jun N-terminal kinase . The generation of H(2)O(2) and activation of caspase 3 were identified as critical components of As(2)O(3)-induced apoptosis in all of the above cell lines . P09038 enhanced As(2)O(3)-induced apoptosis in JK-GMS cells . The overall effects of As(2)O(3) strongly suggest that it has therapeutic potential for the treatment of ES/PNET . DB01037 transdermal system : in the treatment of major depressive disorder . The monamine oxidase ( MAO ) inhibitor selegiline is selective for P27338 at the low oral dosages used in the treatment of Parkinson 's disease . However , P21397 is also inhibited at the high oral dosages needed to effectively treat depression ( not an approved indication ) , necessitating a tyramine-restricted diet . The selegiline transdermal system was designed to deliver antidepressant drug concentrations to the CNS , without substantially impairing small intestine P21397 activity . At the target dose of 6 mg/24 hours , tyramine dietary restrictions are not needed . Short-term treatment with fixed ( 6 mg/24 hours ) or flexible ( 6 , 9 or 12 mg/24 hours ) doses of selegiline transdermal system was superior to placebo on most measures of antidepressant activity in 6- or 8-week , randomised , double-blind , multicentre studies in adult outpatients with major depressive disorder ( MDD ) . Likewise , long-term treatment with a fixed dose of selegiline transdermal system 6 mg/24 hours was superior to placebo as maintenance therapy in a 52-week , randomised , double-blind , multicentre , relapse-prevention trial in patients with MDD . DB01037 transdermal system therapy was generally well tolerated in placebo-controlled studies ; application site reactions , mostly of mild to moderate severity , were the most commonly reported adverse events . The incidence of sexual adverse effects and weight gain was low and similar to that with placebo . DB00293 ( DB00293 ) : from concept to care , a programme in rational drug discovery . DB00158 -based anticancer drugs with specificity for thymidylate synthase ( TS ) have come of age . Ideas nurtured in the early 1970s led to the first-generation of antifolates with TS and dihydrofolate reductase ( P00374 ) inhibitory activities . Compounds with increased selectivity for TS followed with the highly specific inhibitor , CB3717 being synthesised in 1979 at the Institute of Cancer Research ( ICR ) . CB3717 had significant clinical activity but its development had to be abandoned because its low aqueous solubility led to occasional nephrotoxicity . Collaborative laboratory studies between the ICR and ICI Pharmaceuticals ( later to become Zeneca Pharmaceuticals ) led to the discovery of DB00293 ( DB00293 ) , the first antifolate to be licensed for the treatment of cancer ( UK 1995 ) in nearly 40 years and the first new drug for colorectal cancer in about 35 years . DB00293 belongs to a class of compounds that use the reduced-folate carrier ( P41440 ) for uptake into cells and which are excellent substrates for folylpolyglutamate synthetase ( Q05932 ) . This paper reviews the underlying philosophies , and the milestones reached during the development of DB00293 . DB01169 phosphorylates c-Fos to transactivate P38936 ( P38936 /CIP1) expression . An infamous poison , arsenic also has been used as a drug for nearly 2400 years ; in recently years , arsenic has been effective in the treatment of acute promyelocytic leukemia . Increasing evidence suggests that opposite effects of arsenic trioxide ( ATO ) on tumors depend on its concentrations . For this reason , the mechanisms of action of the drug should be elucidated , and it should be used therapeutically only with extreme caution . Previously , we demonstrated the opposing effects of P27361 /2 and JNK on P38936 ( P38936 /CIP1) ( P38936 ) expression in response to ATO in A431 cells . In addition , JNK phosphorylates c-Jun ( DB00133 (63/73) ) to recruit Q15583 / Q13547 to suppress P38936 gene expression . Presently , we demonstrated that a high concentration of ATO sustains P27361 /2 phosphorylation , and increases c-Fos biosynthesis and stability , which enhances P38936 gene expression . Using site-directed mutagenesis , a DNA affinity precipitation assay , and functional assays , we demonstrated that phosphorylation of the C-terminus of c-Fos ( DB00156 (232) , DB00156 (325) , DB00156 (331) , and DB00133 (374) ) plays an important role in its binding to the P38936 promoter , and in conjunction with N-terminus phosphorylation of c-Fos ( DB00133 (70) ) to transactivate P38936 promoter expression . In conclusion , a high concentration of ATO can sustain P27361 /2 activation to enhance c-Fos expression , then dimerize with dephosphorylated c-Jun ( DB00133 (63/73) ) and recruit p300/CBP to the Sp1 sites ( -84/-64 ) to activate P38936 gene expression in A431 cells . P10275 is expressed in murine choroid plexus and downregulated by 5alpha-dihydrotestosterone in male and female mice . The choroid plexuses ( CPs ) of the brain form a unique interface between the peripheral blood and the cerebrospinal fluid ( P04141 ) . CPs produce several neuroprotective peptides , which are secreted into the P04141 . Despite their importance in neuroprotection , the mechanisms underlying the regulation of most of these peptides in CPs remain unknown . Androgens regulate the expression of neuroprotective peptides in several tissues where the androgen receptor ( AR ) is coexpressed , including the brain . The presence of AR in CPs has never been investigated , but recent studies in our laboratory show that the CP is an androgen-responsive tissue . In order to fulfill this gap , we investigated and characterized AR distribution and expression in male and female rat CPs and in primary cultures from rat CP epithelial cells . In addition , the response of AR to 5alpha-dihydrotestosterone ( DB02901 ) in castrated male and female mice subjected to DB02901 replacement was analyzed . We show that rat CP epithelial cells contain AR mRNA and protein . Moreover , we demonstrate that AR is downregulated by DB02901 in mice CPs . Copy number analysis of 24 oncogenes : O15151 identified as a putative marker for low recurrence risk in non muscle invasive bladder cancer . Patients with non-muscle invasive bladder cancer ( NMIBC ) generally have a high risk of relapsing locally after primary tumor resection . The search for new predictive markers of local recurrence thus represents an important goal for the management of this disease . We studied the copy number variations ( CNVs ) of 24 oncogenes ( O15151 , P04198 , Q9UM73 , P16234 , P10721 , P35968 , P00374 , P00533 , MET , SMO , P11362 , MYC , P00519 , P07949 , P24385 , P30279 , P11802 , Q00987 , Q96GD4 , P04626 , P11388 , O14965 , AR and P15056 ) using multiplex ligation probe amplification technique to verify their role as predictive markers of recurrence . DB03843 -fixed paraffin-embedded tissue samples from 43 patients who underwent transurethral resection of the bladder ( TURB ) were used ; 23 patients had relapsed and 20 were disease-free after 5 years . Amplification frequencies were analyzed for all genes and O15151 was the only gene that showed significantly higher amplification in non recurrent patients than in recurrent ones ( 0.65 vs. 0.3 ; Fisher 's test p=0.023 ) . Recurrence-free survival analysis confirmed the predictive role of O15151 ( log-rank test p=0.041 ) . Our preliminary results indicate a putative role for the O15151 gene in predicting local recurrence of bladder cancer . Confirmation of this hypothesis is needed in a larger cohort of NMIBC patients . Multiple pathways of apolipoprotein E signaling in primary neurons . P02649 is a genetic risk factor for Alzheimer 's disease , and the apoE protein is associated with beta-amyloid deposits in Alzheimer 's disease brain . We examined signaling pathways stimulated by apoE in primary neurons in culture . ApoE and an apoE-derived peptide activated several intracellular kinases , including prominently extracellular signal-regulated kinase 1/2 ( P27361 /2 ) . P27361 /2 activation by apoE was blocked by an inhibitor of the low-density lipoprotein receptor family , the specific DB01221 glutamate receptor antagonist MK 801 and other calcium channel blockers . Activation of apoE receptors also induced tyrosine phosphorylation of Dab1 , an adaptor protein of apoE receptors , but experiments in Dab1 knockout neurons demonstrated that Dab1 was not necessary for P29323 activation . In contrast , apoE treatment of primary neurons decreased activation of c-Jun N-terminal kinase , a kinase that interacts with another apoE receptor adaptor protein , c-Jun N-terminal kinase-interacting protein . This change also depended on interactions with the low-density lipoprotein receptor family but was independent of calcium channels . c-Jun N-terminal kinase deactivation by apoE was blocked by gamma-secretase inhibitors and pertussis toxin . These results demonstrate that apoE affects several signaling cascades in neurons : increased disabled phosphorylation , activation of the P27361 /2 pathway ( dependent on calcium influx via the DB01221 receptor ) and inhibition of the P45983 /2 pathway ( dependent on gamma-secretase and G proteins ) . Convergent and divergent cellular responses by ErbB4 isoforms in mammary epithelial cells . Associations of ErbB4 ( Q15303 / Q15303 ) , the fourth member of the P00533 family , with cancer are variable , possibly as a result of structural diversity of this receptor . There are multiple structural isoforms of Q15303 arising by alternative mRNA splicing , and a subset undergo proteolysis that releases membrane-anchored and soluble isoforms that associate with transcription factors and coregulators to modulate transcription . To compare the differential and common signaling activities of full-length ( FL ) and soluble intracellular isoforms of Q15303 , four JM-a isoforms ( FL and soluble intracellular domain ( ICD ) CYT-1 and CYT-2 ) were expressed in isogenic MCF10A cells and their biologic activities were analyzed . Both FL and ICD CYT-2 promoted cell proliferation and invasion , and CYT-1 suppressed cell growth . Transcriptional profiling revealed several new and underexplored Q15303 -regulated transcripts , including : proteases/protease inhibitors ( P08254 and P07093 ) , the YAP/Hippo pathway ( P29279 , O00622 , and P09486 ) , the mevalonate/cholesterol pathway ( P04035 , Q01581 , P01130 , and Q9UBM7 ) , and cytokines ( P10145 , P78556 , and P09341 ) . Many of these transcripts were subsequently validated in a luminal breast cancer cell line that normally expresses Q15303 . Furthermore , ChIP-seq experiments identified O75689 , P02649 , P09486 , P16949 , and Q05195 as novel molecular targets of Q15303 . These findings clarify the diverse biologic activities of Q15303 isoforms , and reveal new and divergent functions . IMPLICATIONS : ErbB4 as a regulator of Hippo and mevalonate pathways provides new insight into milk production and anabolic processes in normal mammary epithelia and cancer . Clot penetration and retention by plasminogen activators promote fibrinolysis . P00750 ( tPA ) remains the sole thrombolytic approved by the FDA for the treatment of pulmonary embolism (PE). tPA has not been replaced by third generation plasminogen activators , e.g. DB00015 ( Ret ) and DB00031 ( TNK ) that circulate with longer life-spans and in theory should have more extended potency in vivo . One reason for this paradox is the inability to assign units of activity to plasminogen activators based on specific biologically relevant standards , which impairs objective comparison . Here , we compare clot permeation , retention and fibrinolytic activities of tPA , TNK and Ret in vitro and clot composition over time with outcome in a mouse model of disseminated pulmonary microembolism ( ME ) . When clots were incubated in the continuous presence of drug , tPA , TNK and Ret lysed fibrin clots identically in the absence of PA inhibitor-1 ( e.g. P05121 ) . Ret , which has lower fibrin affinity and greater susceptibility to inhibition by P05121 than tPA , was less effective in lysing plasma clots , while TNK was less effective when the fibrin content of the clots was enhanced . However , when clots were afforded only brief exposure to drug , as occurs in vivo , Ret showed more extensive clot permeation , greater retention and lysis than tPA or TNK . These results were reproduced in vivo in a mouse model of ME . These studies indicate the need for more relevant tests of plasminogen activator activity in vitro and in vivo and they show that clot permeation and retention are important potential predictors of clinical utility . Connexin 43 and P29323 regulate tension-induced signal transduction in human periodontal ligament fibroblasts . Periodontal ligament ( PDL ) fibroblasts play an important role in preserving periodontal homeostasis and transmitting mechanical signals to alveolar bone . Connexin 43 ( P17302 ) , a gap junction protein , is essential for bone homeostasis and regulates bone remodeling . However , the function of P17302 in human PDL fibroblast-regulated bone remodeling has not yet been elucidated . In this study , human PDL fibroblasts were exposed to cyclic mechanical tension with a maximum 5 % elongation for different durations . We then examined the expression of signaling molecules related to osteogenesis and osteoclastogenesis at both the mRNA and protein levels as well as the activity of extracellular signal-regulated kinase ( P29323 ) in human PDL fibroblasts after loading . We found that mechanical tension increased P17302 , which further upregulated osteogenic ( e.g. , Q13950 , Osterix , and O00300 ) and down-regulated osteoclastogenic ( e.g. , O14788 ) signaling molecules . Suppressing P17302 gene ( Gja1 ) by siRNA inhibited the increase in osteogenesis-related molecules but enhanced O14788 expression . Similar to P17302 , activated P27361 /2 was also enhanced by mechanical tension and suppressed by P17302 siRNA . Inhibition of P27361 /2 signaling using PD98059 reduced the tension-regulated increase in osteogenesis-related molecules but enhanced that of osteoclastogenesis-related ones . These findings suggest that cyclic tension may involve into the osteogenic or osteoclastogenetic differentiation potential of human PDL fibroblasts via the P17302 - P27361 /2 signaling pathway . Regulatory roles of sex hormones in cutaneous biology and immunology . Recent studies have revealed that sex hormones manifest a variety of biological and immunological effects in the skin . Pregnancy , menstruation and the menopause modulate the natural course of psoriasis , indicating a female hormone-induced regulation of skin inflammation . Estrogen in vitro down-regulates the production of the neutrophil , type 1 T cell and macrophage-attracting chemokines , P10145 , P02778 , P13501 , by keratinocytes , and suppresses IL-12 production and antigen-presenting capacity while enhancing anti-inflammatory P22301 production by dendritic cells . These data indicate that estrogen may attenuate inflammation in psoriatic lesions . Estrogen , alone or together with progesterone , prevents or reverses skin atrophy , dryness and wrinkles associated with chronological or photo-aging . Estrogen and progesterone stimulate proliferation of keratinocytes while estrogen suppresses apoptosis and thus prevents epidermal atrophy . Estrogen also enhances collagen synthesis , and estrogen and progesterone suppress collagenolysis by reducing matrix metalloproteinase activity in fibroblasts , thereby maintaining skin thickness . Estrogen maintains skin moisture by increasing acid mucopolysaccharide or hyaluronic acid levels in the dermis . Progesterone increases sebum secretion . Estrogen accelerates cutaneous wound healing stimulating P01138 production in macrophages , GM- P04141 production in keratinocytes and P09038 and TGF-beta1 production in fibroblasts , leading to the enhancement of wound re-innervation , re-epithelialization and granulation tissue formation . In contrast , androgens prolong inflammation , reduce deposition of extracellular matrix in wounds , and reduce the rate of wound healing . Estrogen enhances P15692 production in macrophages , an effect that is antagonized by androgens and which may be related to the development of granuloma pyogenicum during pregnancy . These regulatory effects of sex steroids may be manipulated as therapeutic or prophylactic measures in psoriasis , aging , chronic wounds or granuloma pyogenicum . In search of the ideal immunopanel to distinguish metastatic mammary carcinoma from primary lung carcinoma : a tissue microarray study of 207 cases . BACKGROUND : Distinguishing metastatic carcinoma of breast origin ( MCBO ) to lung from primary lung carcinomas ( P98160 ) is a diagnostic quandary with clinical ramifications . Immunostains CK7 , CK20 , ER , PR , and Mammaglobin as well as pertinent negative stains are utilized but prove insufficient . We set out to identify stains either alone or as a group that would better discern between these 2 entities . DESIGN : Tissue microarrays of 109 MCBO to lung and 102 P98160 were stained with CK7 , CK20 , ER , PR , AR , Mammaglobin , Napsin A , GATA-3 , and Q15669 -1 . An H-score was calculated for each case and stain . RESULTS : The highest area under the receiver-operating characteristic curves for each stain was seen with GATA-3 ( 0.817 ) , Napsin A ( 0.817 ) , and Q15669 -1 ( 0.854 ) . When all possible combinations were analyzed , GATA-3 and Q15669 -1 proved to correctly classify with the highest accuracy ( 0.934 ) . Combinations of GATA-3 and Napsin A ( 0.920 ) and GATA-3 , Napsin A , and Q15669 -1 ( 0.933 ) were not significantly different from GATA-3 and Q15669 -1 . The odds ratios for each stain and combination of stains showed that those for GATA-3 and Q15669 -1 were divergent , signifying that cases with higher H-scores for GATA-3 and Q15669 -1 were more likely to be classified as MCBO and P98160 , respectively . CONCLUSIONS : GATA-3 and Q15669 -1 can correctly classify a case as either MCBO or P98160 in 93.4 % of cases . Although highly specific and sensitive for P98160 , Napsin A in lieu of Q15669 -1 or as an additional stain did not improve classification accuracy . Genotype frequencies of 50 polymorphisms for 241 Japanese non-cancer patients . This paper lists the genotype frequencies of 50 polymorphisms of 37 genes ( P05091 , P07550 , P13945 , P21964 , P16671 , P25025 , P24385 , P35354 , P11509 , P05093 , P11511 , IGF1 , IL-1A , IL-1B , IL-1RN , IL-1R1 , P05231 , P10145 , P22301 , P41159 , Le , L-myc , P05164 , Q99707 , P42898 , P21397 , P15559 , O15527 , p53 , p73 , Se , P31213 , TGF-B , P01375 -A , P01375 -B , P18074 , and P18887 ) and 6 sets of combined genotype frequencies for 241 non-cancer Japanese outpatients . Though the genotype frequencies of 25 polymorphisms have already been reported in our previous papers , 15 polymorphisms ( P16671 A52C , P25025 C785T , P24385 G870A , IGF1 C/T at intron 2 and G2502T , IL-1A 46-bp VNTR , IL-1R1 C-116T , P05231 Ins/Del 17C , P10145 A-278T and C74T , IL- 10 T-819C , P41159 A-2548G , P31213 2-bp VNTR , P18074 Lys751Gln , and P18887 Arg399Gln ) and six sets of combined genotype frequencies ( IL-1B C-31T and IL-1A C-889T , IL-1B C-31T and IL-1RN 86-bp VNTR , IL-1B C-31T and IL-1R1 C-116T , P01375 -A G-308A and P01375 -B A252G , P31213 Val89Leu and 2-bp VNTR , and P18887 Arg399Gln and P18074 Lys751Gln ) were reported in this paper for the first time for Japanese . Although microarray technology will produce this kind of information in near future , this is the first document that reports the genotype/allele frequencies among Japanese for an archival purpose . Somatostatin preserved blood brain barrier against cytokine induced alterations : possible role in multiple sclerosis . Multiple sclerosis ( MS ) is an inflammatory neurological disorder associated with demyelination , impaired blood brain barrier ( BBB ) , axonal damage and neuronal loss . In the present study , we measured somatostatin ( P61278 ) and tumor necrosis factor-α ( P01375 -α ) like immunoreactivity in P04141 samples from MS and non-MS patients . We also examined the role of P61278 in cytokines and lipopolysaccharide ( LPS ) -induced damage to the BBB using human brain endothelial cells in culture . Most of the cerebrospinal fluid ( P04141 ) samples studied from definite MS patients exhibited lower somatostatin ( P61278 ) -like immunoreactivity and higher expression of P01375 -α in comparison to non-MS patients . Treatment of cells with cytokines and LPS blocked P61278 secretion and decreased P61278 expression . Human brain endothelial cells expressed all five somatostatin receptors ( SSTRs ) with increased expression of P30874 and 4 upon treatment with cytokines and LPS . Cytokines and LPS-induced disruption of the tight junction proteins Zonula occludens ( ZO-1 ) organization was restored in presence of P61278 , P30874 or P31391 selective agonists . Furthermore , inflammation induced changes in extracellular signal-regulated kinases ( P27361 /2 and Q13164 ) signaling and altered expression of endothelial and inducible nitric oxide synthase are modulated in presence of P61278 . These data indicate that decreased levels of P61278 contribute to failure of the BBB in MS . DB00104 and the novel multireceptor ligand somatostatin receptor agonist pasireotide ( DB06663 ) block the adrenalectomy-induced increase in mitotic activity in male rat anterior pituitary . The novel somatostatin receptor agonist pasireotide binds with high affinity to somatostatin receptors P30872 , 2 , 3 , and 5 . Acting principally through the latter , it inhibits basal and P06850 -stimulated DB01285 secretion from the AtT20 corticotroph cell line and DB01285 release from a proportion of human corticotroph adenomas both in vitro and in vivo . Data supporting an additional antiproliferative effect has led to pasireotide being explored as a potential therapy for patients with Cushing 's disease . We have compared the effects of pasireotide and octreotide on adrenalectomy-induced mitotic and apoptotic activity in the male rat anterior pituitary . Adrenalectomized rats were treated with daily sc injections of vehicle , pasireotide , or octreotide . Changes in proliferation and apoptosis were determined 2-6 d postoperatively . DB06663 and octreotide had no effect on baseline pituitary cell turnover and no measurable effects on apoptosis . However , the wave of increased mitotic activity normally seen in the pituitary after adrenalectomy was completely abolished . Nevertheless , pasireotide and octreotide did not diminish the increase in DB01285 -immunopositive cell index after adrenalectomy , indicating that cell division and differentiation of hormonally null cells in the pituitary are under independent control . In conclusion , basal cell turnover in the pituitary is not inhibited by pasireotide or octreotide . Bilateral adrenalectomy stimulates differentiation of preexisting null cells into DB01285 -positive cells . Cell division after bilateral adrenalectomy occurs in a specific subpopulation of hormonally null cells that are equally sensitive to the antiproliferative effects of pasireotide and octreotide , implicating P30874 receptors in this antimitotic response . Q96RP3 is expressed in human pregnant myometrial cells and regulates myosin light chain phosphorylation : potential role of the type-2 corticotropin-releasing hormone receptor in the control of myometrial contractility . The family of P06850 -related peptides are suggested to play important roles in the control of myometrial contractility during pregnancy and labor . In this study we investigated the expression of urocortin II ( P55089 II ) in human myometrium and its ability to phosphorylate intracellular components that can be involved in modulating myometrial contractility . Using RT-PCR and fluorescent in situ hybridization , we demonstrated that P55089 II and type-2 P06850 receptor ( Q13324 ) mRNAs were expressed in human nonpregnant and pregnant myometrium . Immunofluorescent studies confirmed protein expression of P55089 II in human pregnant myometrial cells , whereas chemical cross-linking studies with radiolabeled P55089 II confirmed the presence of Q13324 sites with an apparent molecular mass of 50 kDa . Treatment of primary human myometrial cells with P55089 II to specifically activate Q13324 resulted in a dose-dependent increase of myosin light chain ( MLC(20) ) phosphorylation . Activation of protein kinase C ( PKC ) and P27361 /2 was required for the P55089 II-induced activation of MLC(20) , because treatment of myometrial cells with inhibitors of MAPK kinase 1 ( U0126 ) and PKC ( bisindolylmaleimide ) inhibited the P55089 II-induced phosphorylation of MLC(20) . Furthermore , the P55089 II effect on MLC(20) was dependent on RhoA translocation to the membrane and subsequent activation of RhoA-associated kinase , as shown by the use of the specific inhibitors exoenzyme P01024 and Y27632 . Collectively , our data suggest a distinctive role for Q13324 - specific agonists like P55089 II in the control of myometrial contractility during human pregnancy involving sequential activation of PKC , MAPK kinase 1 , P27361 /2 , RhoA , and RhoA-associated kinase , leading to the MLC(20) phosphorylation . DB00104 prevents growth factor-induced proliferation of bovine retinal endothelial cells under hypoxia . Ocular diseases such as proliferative diabetic retinopathy are the major cause of blindness in industrialized countries . The main pathologic features of these diseases are hypoxia and overproduction of growth factors resulting in pathologic proliferation of endothelial cells and new vessel formation . Particularly , hypoxia and growth factors , such as P15692 , DB01277 , P09038 and TGF beta(2) , show a complex interaction in the onset and progression of the diseases . Therefore , to date , most therapeutic strategies for proliferative retinopathies have targeted proliferation of endothelial cells evoked by growth factors . Recently , a synthetic analog of somatostatin , octreotide , has been shown to inhibit the proliferation of various cells in vitro , including endothelial cells . In this study , we have investigated the proliferative response of bovine retinal endothelial cells ( BREC ) to growth factors under hypoxic conditions and the modulation by octreotide . We found a dose-dependent increase of cell proliferation with P15692 , DB01277 and P09038 , and inhibition of hypoxia-induced cell proliferation with TGF beta(2) . Moreover , growth factor-induced , but not hypoxia-induced , cell proliferation was attenuated in the presence of octreotide . In contrast , TGF beta(2) abolished hypoxia-induced BREC proliferation . Similar to octreotide BIM23027 , a somatastatin receptor subtype 2 ( P30874 ) receptor agonist was able to attenuate the growth factor-induced proliferation of BREC expressing mRNA and protein for P30874 . Therefore , we postulate that octreotide exerts its effects mainly through binding to the P30874 . Taken together , our findings point to octreotide as a promising candidate for the treatment of eye disorders involving growth factor-dependent proliferation of endothelial cells . DB01169 -mediated antiplatelet activity : pivotal role of the phospholipase C gamma 2-protein kinase C-p38 MAPK cascade . DB01169 produces high rates of complete clinical remission in patients with relapsed/refractory acute promyelocytic leukemia . Platelet activation is relevant in a variety of acute thrombotic events and coronary heart diseases . Few studies have examined the effects of arsenic trioxide on platelets , and the mechanisms underlying the signaling pathways remain obscure . The aim of this study was to examine systematically the detailed mechanisms of arsenic trioxide in preventing platelet activation . DB01169 ( 5 micromol/L ) exhibited more potent activity at inhibiting collagen ( 1 microg/mL ) -induced platelet aggregation than other agonists . DB01169 ( 15 and 25 micromol/L ) inhibited collagen-induced platelet activation accompanied by [Ca(+2)]i mobilization , thromboxane A(2) ( TxA(2) ) formation , phospholipase C ( P98160 )gamma 2 phosphorylation , and protein kinase C ( PKC ) activation . DB01169 ( 15 and 25 micromol/L ) did not significantly affect cyclic nucleotide-induced vasodilator-stimulated phosphoprotein phosphorylation . Moreover , arsenic trioxide markedly inhibited p38 mitogen-activated protein kinase ( MAPK ) but not P45983 /2 or P28482 phosphorylation in washed platelets . DB01169 also markedly reduced hydroxyl radical ( OH(.) ) formation in the erythrocyte sedimentation rate ( P03372 ) study . The most important findings of this study suggest that the inhibitory effect of arsenic trioxide possibly involves inhibition of the P98160 gamma 2-PKC-p38 MAPK cascade , thereby leading to inhibition of [Ca(+2)]i or free radical formation , and finally the inhibition of platelet aggregation . DB01169 induces apoptosis through JNK and P29323 in human mesothelioma cells . Malignant mesothelioma is an aggressive tumor of serosal surfaces , which is refractory to current treatment options . DB01169 ( As2O3 ) is used clinically to treat acute promyelocytic leukemia , and also to inhibit proliferation of several solid tumors including hepatoma , esophageal , and gastric cancer in vitro . Here we found that As2O3 inhibited cell viability of a mesothelioma cell line , NCI-H2052 . As2O3 induced apoptosis of NCI-H2052 cells , which was accompanied by activation of c-Jun NH2-terminal kinase (JNK)1/2 , extracellular signal-regulated kinase ( P29323 )1/2 , and caspase-3 . zVAD-fmk , a broad-spectrum caspase inhibitor , inhibited As2O3-induced apoptosis and activation of caspase-3 , but not that of P45983 /2 and P27361 /2 . Small interfering RNAs ( siRNAs ) targeting P45983 /2 suppressed As2O3-induced caspase-3 activation and apoptosis , indicating that P45983 /2 regulate As2O3-induced apoptosis though caspase cascade . Furthermore , P45983 siRNA abrogated As2O3-induced P45984 phosphorylation and P45984 siRNA abrogated As2O3-induced P45983 phosphorylation , suggesting that P45983 and P45984 interact with each other . Moreover , P45983 siRNA , but not P45984 siRNA , abrogated As2O3-induced P27361 /2 phosphorylation . P45984 siRNA together with PD98059 , a specific MAPK/ P29323 kinase inhibitor , suppressed As2O3-induced apoptosis more significantly than P45984 siRNA alone . These results indicated that As2O3 induces apoptosis of NCI-H2052 cells mainly through P45983 /2 activation , and that P27361 /2 is involved in As2O3-induced apoptosis when P45983 /2 are inactivated . Interaction of the cholinergic system and the hypothalamic-pituitary-adrenal axis as a risk factor for depression : evidence from a genetic association study . Chronic stress and related disruption of hypothalamic-pituitary-adrenal axis reactivity is a known risk factor for depression . Besides its effects on glucocorticoids , stress also impacts the cholinergic system . Therefore , the interaction of two polymorphisms , one on the cholinergic P43681 receptor gene and one on the glucocorticoid receptor gene ( P04150 ) , on depression was investigated . In a sample of 800 healthy participants , we genotyped for the P24385 rs41423247 and the P43681 rs1044396 single-nucleotide polymorphisms and assessed depressiveness by means of the Beck Depression Inventory . We identified a significant epistasis effect P24385 by P43681 showing that carriers of the CC genotype at the P24385 locus who were also homozygous for the T allele at the P43681 locus had the highest depression scores . This is the first evidence from molecular genetics to show that the hypothalamic-pituitary-adrenal axis and the cholinergic system -- both involved in stress reactivity -- represent a combined risk factor for depression . Importance of MEK in neutrophil microbicidal responsiveness . Exposure of neutrophils to inflammatory stimuli such as the chemoattractant FMLP leads to activation of responses including cell motility , the oxidative burst , and secretion of proteolytic enzymes . A signaling cascade involving sequential activation of P04049 , mitogen-activated protein kinase ( MEK ) , and extracellular signal regulated kinase ( P29323 ) is also rapidly activated after agonist exposure . The temporal relationship between these events suggests that the kinases may be involved in triggering the effector functions , but direct evidence of a causal relationship is lacking . To assess the role of the MEK/ P29323 pathway in the activation of neutrophil responses , we studied the effects of PD098059 , a potent and selective inhibitor of MEK . Preincubation of human neutrophils with 50 microM PD098059 almost completely ( > 90 % ) inhibited the FMLP-induced activation of MEK-1 and MEK-2 , the isoforms expressed by neutrophils . This dose of PD098059 virtually abrogated chemoattractant-induced tyrosine phosphorylation and activation of P27361 and P28482 , implying that MEKs are the predominant upstream activators of these mitogen-activated protein kinases . Pretreatment of neutrophils with the MEK antagonist inhibited the oxidative burst substantially and phagocytosis only moderately . In addition , PD098059 antagonized the delay of apoptosis induced by exposure to granulocyte-macrophage P04141 . However , the effects of PD098059 were selective , as it failed to inhibit other responses , including chemoattractant-induced exocytosis of primary and secondary granules , polymerization of F-actin , chemotaxis , or activation of phospholipase A2 . We conclude that MEK and P29323 contribute to the activation of the oxidative burst and phagocytosis , and participate in cytokine regulation of apoptosis . Menadione reduction by pharmacological doses of ascorbate induces an oxidative stress that kills breast cancer cells . Oxidative stress generated by ascorbate-driven menadione redox cycling kills MCF7 cells by a concerted mechanism including glycolysis inhibition , loss of calcium homeostasis , DNA damage and changes in mitogen activated protein kinases ( MAPK ) activities . Cell death is mediated by necrosis rather than apoptosis or macroautophagy . Neither 3-methyladenine nor Z-VAD affects cytotoxicity by ascorbate/menadione ( Asc/Men ) . BAPTA-AM , by restoring cellular capacity to reduce MTT , underlines the role of calcium in the necrotic process . Oxidative stress-mediated cell death is shown by the opposite effects of DB06151 and 3-aminotriazole . Moreover , oxidative stress induces DNA damage ( protein poly-ADP-ribosylation and gamma- P16104 phosphorylation ) and inhibits glycolysis . Asc/Men deactivates extracellular signal-regulated kinase ( P29323 ) while activating p38 , suggesting an additional mechanism to kill MCF7 cells . Since ascorbate is taken up by cancer cells and , due to their antioxidant enzyme deficiency , oxidative stress should affect cancer cells to a greater extent than normal cells . This differential sensitivity may have clinical applications . Multiplex protein signature for the detection of bladder cancer in voided urine samples . PURPOSE : Accurate urine assays for bladder cancer detection would benefit patients and health care systems . Through extensive genomic and proteomic profiling of urine components we previously identified a panel of 8 biomarkers that can facilitate the detection of bladder cancer in voided urine samples . In this study we confirmed this diagnostic molecular signature in a diverse multicenter cohort . MATERIALS AND METHODS : We performed a case-control , phase II study in which we analyzed voided urine from 102 subjects with bladder cancer and 206 with varying urological disorders . The urinary concentration of 8 biomarkers ( P10145 , P14780 and 10 , P05121 , P15692 , P03950 , Q16790 and P02649 ) was assessed by enzyme-linked immunosorbent assay . Diagnostic performance of the panel of tested biomarkers was evaluated using ROCs and descriptive statistical values , eg sensitivity and specificity . RESULTS : Seven of the 8 urine biomarkers were increased in subjects with bladder cancer relative to those without bladder cancer . The 7 biomarkers were assessed in a new model , which had an AUROC of 0.88 ( 95 % CI 0.84-0.93 ) , and 74 % sensitivity and 90 % specificity . In contrast , the sensitivity of voided urine cytology and the UroVysion® cytogenetic test in this cohort was 39 % and 54 % , respectively . Study limitations include analysis performed on banked urine samples and the lack of voided urine cytology and cytogenetic test data on controls . CONCLUSIONS : The study provides further evidence that the reported panel of diagnostic biomarkers can reliably achieve the noninvasive detection of bladder cancer with higher sensitivity than currently available urine based assays . Large-cell neuroendocrine carcinoma of the ampulla of Vater . Large-cell neuroendocrine carcinoma is a high-grade neuroendocrine carcinoma , originally described in the lung . The tumor rarely occurs in extrapulmonary sites like the gastrointestinal tract , and only few examples have been described in the ampulla of Vater . A new case of large-cell neuroendocrine carcinoma of the ampulla of Vater in a 60-year-old man is reported . After pancreatoduodenectomy , macroscopic examination revealed ulcerated tumor in the region of the ampulla of Vater . Microscopically , the tumor exhibited organoid , predominantly nested growth pattern , consisting of large , polygonal cells with pleomorphic nuclei . Average number of mitoses was 36 per 10 high-power fields . Small and large areas of necrosis were identified . Immunohistochemically , the tumor cells were positive for synaptophysin , chromogranin A , P09936 , neuron-specific enolase , pancytokeratin , CK8 and somatostatin and negative for CK7 , CK20 , S-100 , Q15669 -1 , HMB-45 , CD117 , P12830 and regulatory peptides . Ki-67 proliferative index was 41 % . Histone deacetylase ( HDAC ) analysis showed almost identical results for Q13547 , Q92769 and O15379 -- 60 , 60.3 and 61 % , respectively . Two months after surgery , liver metastases occurred , confirming highly aggressive behavior of large-cell neuroendocrine carcinoma . DB01169 induces cardiac fibroblast apoptosis in vitro and in vivo by up-regulating TGF-β1 expression . DB01169 ( As2O3 ; ATO ) is clinically effective in treating acute promyelocytic leukemia ( APL ) ; however , it frequently causes cardiotoxic effects . This study was designed to investigate whether ATO could induce apoptosis of cardiac fibroblasts ( CFs ) that play very important roles in maintaining the structure integrity and function of the heart . Cardiac fibroblasts from guinea pigs administered with ATO ( 1mg/kgbw ) were used to test the pro-apoptotic role of ATO in vivo . The current study demonstrated that ATO induced morphological characteristics of apoptosis and P42574 activation in CFs of guinea pigs along with a significant up-regulation in TGF-β1 protein expression , Bax/Bcl-2 ratio and P27361 /2 phosphorylation . In vitro MTT assay showed that ATO remarkably reduced the viability of cultured cardiac fibroblasts ( NRCFs ) from neonatal rat in a concentration- and time-dependent manner . Consistent with the notions in vivo , ATO significantly induced the apoptosis in NRCFs , dramatically up-regulated TGF-β1 protein level and Bax/Bcl-2 ratio in a time-dependent fashion and activated P42574 and P27361 /2 . Finally , pretreatment with LY364947 , an inhibitor of TGF-β signaling could apparently reverse these changes . We therefore conclude that TGF-β is functionally linked to P27361 /2 and that TGF-β signaling is responsible for ATO-induced CFs apoptosis , which provides a novel mechanism of ATO related cardiac toxicology . DB02901 interacts with P00533 /MAPK signalling and modulates P00533 levels in androgen receptor-positive LNCaP prostate cancer cells . P10275 ( AR ) signalling plays a pivotal role in prostate cancer pathogenesis and progression . However , androgen-mediated AR signalling is yet to be fully understood . P00533 and Q96HU1 kinase signalling pathways play predominant roles in AR function . Therefore , we investigated the interaction of P00533 signalling and AR activity in AR-positive LNCaP cells . We found that 5alpha-dihydrotestosterone ( DB02901 ) and P01133 had a synergistic effect on AR activity as detected by a luciferase reporter system , although P01133 alone did not activate AR . Both P27361 /2 and p38 were involved in DB02901 and DB02901 / P01133 -induced AR activation as detected by specific MEK and p38 inhibitors . Furthermore , 24-h treatment of the cells with DB02901 resulted in ubiquitination and down-regulation of the P00533 . This effect could be inhibited by the anti-androgen flutamide , suggesting an androgen-dependent mechanism . On the other hand , DB02901 -treatment strongly increased AR levels in LNCaP cells . These data suggest a complex regulatory loop between activated AR and P00533 . In conclusion , activation of AR by both DB02901 and P01133 / DB02901 involves the Q96HU1 kinase pathway . Long-term activation of AR results in increase of AR levels , which through so far unknown regulatory mechanisms results in ubiquitination and degradation of the P00533 . Is phentermine an inhibitor of monoamine oxidase ? A critical appraisal . DB00191 produces a spectrum of concentration-dependent biochemical effects . It interacts with NE transporters at 0.1 microM , DA transporters at about 1 microM , 5-HT transporters at 15 microM and P21397 at about 100 microM . When administered at typical anorectic doses , phentermine primarily interacts with DA and NE transporters and does not produce biochemical or neurochemical effects which would occur if it were inhibiting P21397 . Some other explanation other than MAO inhibition must be sought to explain how oral phentermine increases platelet 5-HT , since platelet P27338 does not metabolize platelet 5-HT , and since amphetamine-type drugs are even weaker inhibitors of P27338 than P21397 . Clinical studies in humans have shown that amphetamine , which is a more potent inhibitor of P21397 than phentermine , does not inhibit P21397 at therapeutic doses . Neither phentermine alone , fluoxetine alone or their combined use have been associated with cardiac valvulopathy , and clinical experience has shown their combined use to be free of significant adverse effects . Viewed collectively , there appears to be no data to support the hypothesis that phentermine inhibits MAO at typical therapeutic doses . Antitumor activity of arsenic trioxide on retinoblastoma : cell differentiation and apoptosis depending on arsenic trioxide concentration . PURPOSE : DB01169 ( ATO ) targets multiple pathways in malignant cells , resulting in the promotion of differentiation or in the induction of apoptosis . The antitumor activity of ATO on retinoblastoma was investigated . METHODS : Human retinoblastoma cells were incubated with various ATO concentrations . The antiproliferative effect of ATO was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay , and the effect of ATO on cell-cycle progression was validated by flow cytometry . At a low concentration , the ATO-induced differentiation of retinoblastoma cells was evaluated by neurofilament expression and extracellular signal-regulated kinase ( P29323 )1/2 activation , which was confirmed by the inhibition of P27361 /2 . At a high concentration , ATO-induced H(2)O(2) production was investigated with the cell-permeable fluorescent dye 2'7'-dichlorofluorescein-diacetate , and the relationship of ATO-induced H(2)O(2) production with caspase-3-dependent apoptosis was validated by Western blot and 4'6-diamidino-2-phenolindole staining , which was confirmed by reactive oxygen species ( ROS ) inhibition . The effect of ATO on tumor formation was assessed with an orthotopic animal model of retinoblastoma . RESULTS : The antitumor activity of ATO in retinoblastoma was related to two main mechanisms , differentiation and apoptosis , which were determined by the level of ATO . At a low dose ( < or= 1 microM ) , ATO induced the differentiation of retinoblastoma cells through P27361 /2 activation , whereas ROS generation by a high dose ( > or= 2 microM ) of ATO induced apoptosis in retinoblastoma cells . Moreover , ATO at low and high doses effectively inhibited tumor formation . CONCLUSIONS : These results suggest that ATO can be used as an effective alternative therapeutic for the treatment of retinoblastoma . Evaluation of neprilysin sequence variation in relation to P04141 β-Amyloid levels and Alzheimer disease risk . P08473 ( NEP ) is a principal peptidase involved in the degradation of β-amyloid ( Aβ ) , and as such its encoding gene ( MME ) has been the target of numerous genetic association studies on Alzheimer disease . Here , in order to attempt replication of previous findings we have investigated several single nucleotide polymorphisms ( SNPs ) that have been claimed to be associated with AD . A key feature of the present study is the complementary investigation of both AD risk and quantitative measures of AD severity , including cerebrospinal ( P04141 ) fluid levels of P05412 -42 . In contrast to the effects of P02649 , none of these measures are detectably influenced by genetic polymorphism in the MME region . We thus , fail to find support for previous results suggesting that MME impacts AD . P00533 regulates osteoclast differentiation and survival through cross-talking with Q9Y6Q6 signaling . The epidermal growth factor receptor ( P00533 ) functions in various cellular physiological processes such as proliferation , differentiation , and motility . Although recent studies have reported that P00533 signaling is involved in osteoclast recruitment and formation , the molecular mechanism of P00533 signaling for the regulation of osteoclastogenesis remains unclear . We investigated the role of the P00533 in osteoclast differentiation and survival and show that the expression of the P00533 was highly up-regulated by receptor activator of nuclear factor-kappaB ligand ( O14788 ) during osteoclast differentiation . P00533 -specific tyrosine kinase inhibitors and P00533 knockdown blocked O14788 -dependent osteoclast formation , suggesting that P00533 signaling plays an important role in osteoclastogenesis . P00533 inhibition impaired the O14788 -mediated activation of osteoclastogenic signaling pathways , including c-Jun N-terminal kinase ( JNK ) , NF-kappaB , and Akt/protein kinase B ( P31749 ) . In addition , P00533 inhibition in differentiated osteoclasts caused apoptosis through caspase activation . Inhibition of the phosphoinositide-3 kinase ( PI3K ) -Akt/ P31749 pathway and subsequent activation of Q92934 and caspases-9 and -3 may be responsible for the P00533 inhibition-induced apoptosis . The P00533 physically associated with receptor activator of nuclear factor-kappaB ( Q9Y6Q6 ) and Grb2-associated binder 2 ( Gab2 ) . Moreover , O14788 transactivated P00533 . These data indicate that P00533 regulates O14788 -activated signaling pathways by cross-talking with Q9Y6Q6 , suggesting that the P00533 may play a crucial role as a Q9Y6Q6 downstream signal and/or a novel type of Q9Y6Q6 co-receptor in osteoclast differentiation and survival . Muscarinic cholinergic signaling in cervical cancer cells affects cell motility via P27361 /2 signaling . AIMS : The etiology of cervical cancer depends primarily on infection with human papillomaviruses , but tobacco smoking is the most important behavioral risk factor for this cancer . Therefore , we have previously confirmed involvement of nicotinic acetylcholine receptors ( nAChRs ) in cervical cancer biology . In order to comprehensively evaluate the role of cholinergic signaling in cervical cells , we have addressed additional participation of muscarinic acetylcholine receptors ( mAChRs ) . MAIN METHODS : We have studied the expression of mAChRs and cholinergic system components by reverse transcription PCR and Western blots , the motility of cervical cancer cells in cell culture , and the signaling from mAChRs via the P27361 /2 signaling pathway . KEY FINDINGS : The cervical cancer cells HeLa , SiHa and CaSki express four of the five mAChRs , M1 , M3 , M4 , and M5 , and the acetylcholine ( ACh ) synthesizing and degrading enzymes choline acetyltransferase ( P28329 ) , acetylcholinesterase ( P22303 ) , and butyrylcholinesterase ( BChE ) , and vesicular ACh transporter ( Q16572 ). mAChR-dependent signaling induces cervical cell motility , which requires P27361 /2 activation , and could be abrogated by mAChR antagonists . SIGNIFICANCE : The epidemiological finding that tobacco smoke raises the prevalence of cervical cancer has led to analysis of the cholinergic signaling in cervical biology and carcinogenesis . Cervical cancer cells express several nAChRs and mAChRs , whose activation leads to changes of cellular properties such as increased motility and proliferation that favor a carcinogenic phenotype . The signaling involves intracellular phosphorylation cascades including P27361 /2 . A new algorithm for weekly phenprocoumon dose variation in a southern Brazilian population : role for P11712 , P08684 /5 and Q9BQB6 genes polymorphisms . DB00946 is widely used in prophylaxis and treatment of thromboembolic disorders . However , its pharmacokinetics and pharmacodynamics vary according to several genetic and non-genetic factors . DB00946 metabolism is mediated by P11712 and CYP3A enzymes . Moreover , Q9BQB6 is phenprocoumon target of action . Therefore , the aim of this study was to evaluate the association of single nucleotide polymorphisms ( SNPs ) in Q9BQB6 , P11712 , P08684 and P20815 genes with the variance of weekly phenprocoumon dose as well as to develop an algorithm for dose prediction based on genetic and environmental factors . A total of 198 patients with stable phenprocoumon dose , 81 % of European ancestry , were investigated . Genotypes were determined by allelic discrimination with TaqMan assays . Polymorphisms -1639G > A and 1173C > T in Q9BQB6 and the presence of P11712 *2 and/or P11712 *3 are associated with lower doses . On the other hand , 3730G > A in Q9BQB6 gene is associated with higher doses . No association was found between P08684 *1B , P20815 *3 and P20815 *6 polymorphisms . Among non-genetic factors , gender , height , age and use of captopril , omeprazole , simvastatin and β-blockers are associated with dose . Two algorithms were derived : one for the whole sample explained 42 % of dose variation and one for patients of European ancestry only which explained 46 % of phenprocoumon dose . The mean absolute difference between observed and predicted dose was low in both models ( 3.92 mg/week and 3.54 mg/week , for models 1 and 2 , respectively ) . However , more studies with other genes and environmental factors are needed to test and to improve the algorithm . DB00184 consumption is regulated by a human polymorphism in dopamine neurons . Smoking is the most important preventable cause of morbidity and mortality worldwide . Recent genome-wide association studies highlighted a human haplotype on chromosome 15 underlying the risk for tobacco dependence and lung cancer . Several polymorphisms in the P32297 - P30532 - P30926 cluster coding for the nicotinic acetylcholine receptor ( nAChR ) α3 , α5 and β4 subunits were implicated . In mouse models , we define a key role in the control of sensitivity to nicotine for the α5 subunit in dopaminergic ( DAergic ) neurons of the ventral tegmental area ( VTA ) . We first investigated the reinforcing effects of nicotine in drug-naive α5(-/-) mice using an acute intravenous nicotine self-administration task and ex vivo and in vivo electrophysiological recordings of nicotine-elicited DA cell activation . We designed lentiviral re-expression vectors to achieve targeted re-expression of wild-type or mutant α5 in the VTA , in general , or in DA neurons exclusively . Our results establish a crucial role for α5*-nAChRs in DAergic neurons . These receptors are key regulators that determine the minimum nicotine dose necessary for DA cell activation and thus nicotine reinforcement . Finally , we demonstrate that a single-nucleotide polymorphism , the non-synonymous α5 variant rs16969968 , frequent in many human populations , exhibits a partial loss of function of the protein in vivo . This leads to increased nicotine consumption in the self-administration paradigm . We thus define a critical link between a human predisposition marker , its expression in DA neurons and nicotine intake . A common haplotype of the nicotine acetylcholine receptor alpha 4 subunit gene is associated with vulnerability to nicotine addiction in men . DB00184 is the major addictive substance in cigarettes , and genes involved in sensing nicotine are logical candidates for vulnerability to nicotine addiction . We studied six single-nucleotide polymorphisms ( SNPs ) in the P43681 gene and four SNPs in the P17787 gene with respect to nicotine dependence in a collection of 901 subjects ( 815 siblings and 86 parents ) from 222 nuclear families with multiple nicotine-addicted siblings . The subjects were assessed for addiction by both the Fagerstrom Test for DB00184 Dependence ( FTND ) and the Revised Tolerance Questionnaire ( RTQ ) . Because only 5.8 % of female offspring were smokers , only male subjects were included in the final analyses ( 621 men from 206 families ) . Univariate ( single-marker ) family-based association tests ( FBATs ) demonstrated that variant alleles at two SNPs , rs1044396 and rs1044397 , in exon 5 of the P43681 gene were significantly associated with a protective effect against nicotine addiction as either a dichotomized trait or a quantitative phenotype ( i.e. , age-adjusted FTND and RTQ scores ) , which was consistent with the results of the global haplotype FBAT . Furthermore , the haplotype-specific FBAT showed a common ( 22.5 % ) P43681 haplotype , GCTATA , which was significantly associated with both a protective effect against nicotine addiction as a dichotomized trait ( Z=-3.04 , P < .005 ) and significant decreases of age-adjusted FTND ( Z=-3.31 , P < .005 ) or RTQ scores ( Z=-2.73 , P=.006 ) . Our findings provide strong evidence suggesting a common P43681 haplotype might be protective against vulnerability to nicotine addiction in men . Gene clustering analysis in human osteoporosis disease and modifications of the jawbone . OBJECTIVE : An analysis of the genes involved in both osteoporosis and modifications of the jawbone , through text mining , using a web search tool , of information regarding gene/protein interaction . DESIGN : The final set of genes involved in the present phenomenon was obtained by expansion-filtering loop . Using a web-available software ( STRING ) , interactions among all genes were searched for , and a clustering procedure was performed in which only high-confidence predicted associations were considered . RESULTS : Two hundred forty-two genes potentially involved in osteoporosis and in modifications of the jawbone were recorded . Seven " leader genes " were identified ( P35222 , P01584 , P05231 , P05412 , Q13950 , P10451 , P01137 ) , while another 10 genes formed the cluster B group ( P12643 , P18075 , P02452 , P05362 , IGF1 , P22301 , P14780 , P19838 , O14788 , P15692 ) . Ninety-eight genes had no interactions , and were defined as " orphan genes " . CONCLUSIONS : The expansion of knowledge regarding the molecular basis causing osteoporotic traits has been brought about with the help of a de novo identification , based on the data mining of genes involved in osteoporosis and in modification of the jawbone . A comparison of the present data , in which no role was verified for 98 genes that had been previously supposed to have a role , with that of the literature , in which another 81 genes , as obtained from GWAS reviews and meta-analyses , appeared to be strongly associated with osteoporosis , probably attests to a lack of information on osteoporotic disease . Overexpression of SnoN/SkiL , amplified at the 3q26.2 locus , in ovarian cancers : a role in ovarian pathogenesis . High-resolution array comparative genomic hybridization of 235 serous epithelial ovarian cancers demonstrated a regional increase at 3q26.2 encompassing SnoN/SkiL , a coregulator of SMAD/TGFbeta signaling . SnoN RNA transcripts were elevated in approximately 80 % of advanced stage serous epithelial ovarian cancers . In both immortalized normal ( TIOSE ) and ovarian carcinoma cell lines ( OVCA ) , SnoN RNA levels were increased by TGFbeta stimulation and altered by LY294002 and JNK II inhibitor treatment suggesting that the PI3K and JNK signaling pathways may regulate TGFbeta-induced increases in SnoN RNA . In TIOSE , SnoN protein levels were reduced 15min post TGFbeta-stimulation , likely by proteosome-mediated degradation . In contrast , in OVCA , SnoN levels were elevated 3h post-stimulation potentially as a result of inhibition of the proteosome . To elucidate the role of SnoN in ovarian tumorigenesis , we explored the effects of both increasing and decreasing SnoN levels . In both TIOSE and OVCA , SnoN siRNA decreased cell growth between 20 and 50 % concurrent with increased P38936 levels . In TIOSE , transient expression of SnoN repressed TGFbeta induction of P05121 promoters with little effect on the P38936 promoter or resultant cell growth . In contrast to the effects of transient expression , stable expression of SnoN in TIOSE led to growth arrest through induction of senescence . Collectively , these results implicate SnoN levels in multiple roles during ovarian carcinogenesis : promoting cellular proliferation in ovarian cancer cells and as a positive mediator of cell cycle arrest and senescence in non-transformed ovarian epithelial cells . P30532 gene D398N polymorphism in Japanese lung adenocarcinoma . BACKGROUND : Recently , to identify genetic factors that modify lung cancer risk , P30532 non-synonymous variant amino acid position 398 ( D398N ) was identified . The site was a highly conserved in the second cellular loop of the nicotinic acetylcholine receptor subunit protein . MATERIALS AND METHODS : We have investigated P30532 gene polymorphism status in 302 surgically treated lung adenocarcinoma cases from Nagoya City University Hospital . The presence or absence of P30532 polymorphism was analyzed by direct sequences . P00533 mutations status was already investigated and reported . RESULTS : We detected nine cases ( 2.98 % ) of P30532 polymorphism ( D398N ) in our cohort . Total P00533 mutations were present in 129 patients ( 42.7 % ) . The polymorphism statuses were not correlated with gender ( women ; 2.1 % versus men ; 3.7 % , P = 0.5119 ) , smoking status ( never smoker ; 2.0 % versus smoker ; 4.0 % , P = 0.3339 ) , pathological stages ( stage I ; 2.6 % versus stage II-IV ; 3.8 % , P = 0.7246 ) , and P00533 mutation status of the lung adenocarcinomas ( mutation ; 2.3 % versus wild type ; 3.7 % , P = 0.7373 ) . In this analysis , P30532 polymorphism ( D398N ) patients had significantly worse prognosis ( 5/9 were dead ; mean survival = 27.1 mo ) than the patients with P30532 wild type ( 74/293 were dead ; mean survival = 113.9 mo ) ( log-rank test ; P = 0.0146 ) . CONCLUSION : Although P30532 polymorphism is rare from Japanese lung cancer , polymorphism status might be correlated with shorter survival . Synergistic inhibition of colon carcinoma cell growth by Hedgehog-Gli1 inhibitor arsenic trioxide and phosphoinositide 3-kinase inhibitor LY294002 . The Hedgehog ( Hh ) signaling pathway not only plays important roles in embryogenesis and adult tissue homeostasis , but also in tumorigenesis . Aberrant Hh pathway activation has been reported in a variety of malignant tumors including colon carcinoma . Here , we sought to investigate the regulation of the Hh pathway transcription factor Gli1 by arsenic trioxide and phosphoinositide 3-kinase ( PI3K ) inhibitor LY294002 in colon carcinoma cells . We transfected cells with siGli1 and observed a significant reduction of Gli1 expression in HCT116 and HT29 cells , which was confirmed by quantitative real-time polymerase chain reaction and Western blots . Knocking down endogenous Gli1 reduced colon carcinoma cell viability through inducing cell apoptosis . Similarly , knocking down Gli2 using short interfering RNA impaired colon carcinoma cell growth in vitro . To elucidate the regulation of Gli1 expression , we found that both Gli inhibitor arsenic trioxide and PI3K inhibitor LY294002 significantly reduced Gli1 protein expression and colon carcinoma cell proliferation . DB01169 treatment also reduced Gli1 downstream target gene expression , such as Bcl2 and P24385 . More importantly , the inhibition of Hedgehog-Gli1 by arsenic trioxide showed synergistic anticancer effect with the PI3K inhibitor LY294002 in colon carcinoma cells . Our findings suggest that the Hh pathway transcription factor Gli1 is involved in the regulation of colon carcinoma cell viability . Inhibition of Hedgehog-Gli1 expression by arsenic trioxide and PI3K inhibitor synergistically reduces colon cancer cell proliferation , indicating that they could be used as an effective anti-colon cancer combination therapy . Analysis of breast cancer related gene expression using natural splines and the Cox proportional hazard model to identify prognostic associations . Many studies correlating gene expression data to clinical parameters assume a linear increase or decrease of the clinical parameter under investigation with the expression of a gene . We have studied genes encoding important breast cancer-related proteins using a model for survival-type data that is based on natural splines and the Cox proportional hazard model , thereby removing the linearity assumption . Expression data of 16 genes were studied in relation to metastasis-free probability in a cohort of 295 consecutive breast cancer patients treated at The Netherlands Cancer Institute . The independent predictive power for disease outcome of the 16 individual genes was tested in a multivariable model with known clinical and pathological risk factors . There is a linear relationship between increasing expression and a higher or lower hazard for distant metastasis for P03372 , Q15303 , P15692 , O96020 , Q15910 , and Q96NZ9 ; for P04626 , P21860 , P24385 , P24864 , O75530 , P61073 , P32248 , P48061 , and P05121 there is no clear increase or decrease ; and for P00533 there seems to be a non-linear relation . Multivariable analysis showed that the 70-gene prognosis profile outperforms all the other variables in the model ( hazard-rate 5.4 , 95 % CI 2.5-11.7 ; P = 0.000018 ) . P00533 -expression seems to have a non-linear relation with disease outcome , indicating that lower but also higher expression of P00533 are associated with worse outcome compared to intermediate expression levels ; the other genes show no or a linear relation . Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 , P21397 , P23560 , NOS3 , P05231 , P12036 , P31645 , P21964 , P48454 and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual 's response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome . SnoN/SkiL expression is modulated via arsenic trioxide-induced activation of the PI3K/AKT pathway in ovarian cancer cells . SnoN/SkiL ( TGFβ regulator ) is dysregulated in ovarian cancer , a disease associated with acquired drug-resistance . DB01169 ( As₂O₃ , used in treating APL ) induces SnoN to oppose the apoptotic response in ovarian cancer cells . We now report that As₂O₃ increases phosphorylation of P00533 /p66ShcA and P00533 degradation . As₂O₃ activates Src(Y416) whose activity ( inhibited by Q99463 ) modulates P00533 activation , its interaction with Shc/Grb2 , and p-AKT . Inhibition of PI3K reduces SnoN and cell survival . Although P00533 or P28482 siRNA did not alter SnoN expression , As₂O₃-induced cleaved PARP was reduced together with increased P98170 . Collectively , As₂O₃ mediates an initial rise in pY-Src(416) to regulate the PI3K/AKT pathway which increases SnoN and cell survival ; these early events may counter the cell death response associated with increased pY- P00533 /MAPK activation . The effect of prenatal nicotine on mRNA of central cholinergic markers and hematological parameters in rat fetuses . A number of studies have demonstrated the influence of nicotine on fetal development . This study determined the expression of choline acetyltransferase ( P28329 ) , vesicular acetylcholine transporter ( Q16572 ) , and high-affinity choline transporter ( Q9GZV3 ) in the forebrain and hindbrain following chronic prenatal nicotine exposure in the rat fetus ( maternal rats were subcutaneously injected with nicotine at different gestation periods ) . We also measured the effect of chronic nicotine exposure on fetal blood pO(2) , pCO(2) , pH , Na(+) and K(+) concentrations , as well as lactic acid levels . Maternal nicotine exposure during pregnancy was associated with a decrease in fetal pO(2) coupled with a significant increase in pCO(2) and lactic acid as well as restricted fetal growth . Additionally , maternal nicotine administration also reduced P28329 , Q16572 , and Q9GZV3 mRNA levels in the fetal brain . DB00184 -induced fetal hypoxic responses and reduced cholinergic marker expression in the brain were more severe when nicotine was started in early gestation . Our results provide new information about the effects of repeated exposure to nicotine in utero on the expression of central P28329 , Q16572 , and Q9GZV3 in the rat fetus . These results indicate that repeated hypoxic episodes or/and a direct effect of nicotine on the central cholinergic system during pregnancy may contribute to brain developmental problems in fetal origin . DB01169 inhibits cell proliferation and human papillomavirus oncogene expression in cervical cancer cells . DB01169 ( As2O3 ) has shown therapeutic effects in some leukemias and solid cancers . However , the molecular mechanisms of its anticancer efficacy have not been clearly elucidated , particularly in solid cancers . Our previous data showed that As2O3 induced apoptosis of human papillomavirus ( HPV ) 16 DNA-immortalized human cervical epithelial cells and cervical cancer cells and inhibited the expression of HPV oncogenes in these cells . In the present study , we systemically examined the effects of As2O3 on five human cervical cancer cell lines and explored the possible molecular mechanisms . MTT assay showed that HPV-negative C33A cells were more sensitive to growth inhibition induced by As2O3 than HPV-positive cervical cancer cells , and HPV 18-positive HeLa and C4-I cells were more sensitive to As2O3 than HPV 16-positive CaSki and SiHa cells . After As2O3 treatment , both mRNA and protein levels of HPV E6 and E7 obviously decreased in all HPV positive cell lines . In contrast , p53 and Rb protein levels increased in all tested cell lines . P05412 protein expression decreased significantly in HeLa , CaSki and C33A cells with ELISA method . These results suggest that As2O3 is a potential anticancer drug for cervical cancer . Genetic markers in the EET metabolic pathway are associated with outcomes in patients with aneurysmal subarachnoid hemorrhage . Preclinical studies show that epoxyeicosatrienoic acids ( EETs ) regulate cerebrovascular tone and protect against cerebral ischemia . We investigated the relationship between polymorphic genes involved in EET biosynthesis/metabolism , cytochrome P450 ( CYP ) eicosanoid levels , and outcomes in 363 patients with aneurysmal subarachnoid hemorrhage ( aSAH ) . Epoxyeicosatrienoic acids and dihydroxyeicosatetraenoic acid ( DHET ) cerebrospinal fluid ( P04141 ) levels , as well as acute outcomes defined by delayed cerebral ischemia ( P42126 ) or clinical neurologic deterioration ( CND ) , were assessed over 14 days . Long-term outcomes were defined by Modified Rankin Scale ( P59665 ) at 3 and 12 months . P10632 *4 allele carriers had 44 % and 36 % lower mean EET and DHET P04141 levels ( P=0.003 and P=0.007 ) and were 2.2- and 2.5-fold more likely to develop P42126 and CND ( P=0.039 and P=0.041 ) , respectively . P34913 55Arg , P51589 *7 , P10632 *1B , and P10632 g.36785A allele carriers had lower EET and DHET P04141 levels . P10632 g.25369T and P10632 g.36755A allele carriers had higher EET levels . Patients with P10632 *2C and P34913 404del variants had worse long-term outcomes while those with P34913 287Gln , P51589 *7 , and P11712 g.816G variants had favorable outcomes . Epoxyeicosatrienoic acid levels were associated with Fisher grade and unfavorable 3-month outcomes . Dihydroxyeicosatetraenoic acids were not associated with outcomes . No associations passed Bonferroni multiple testing correction . These are the first clinical data demonstrating the association between the EET biosynthesis/metabolic pathway and the pathophysiology of aSAH . DB03419 incorporation into genomic DNA does not predict toxicity caused by chemotherapeutic inhibition of thymidylate synthase . P04818 ( TS ) is an important target of several chemotherapeutic agents , including DB00544 and raltitrexed ( DB00293 ) . During TS inhibition , TTP levels decrease with a subsequent increase in dUTP . DB03419 incorporated into the genome is removed by base excision repair ( BER ) . Thus , BER initiated by uracil DNA glycosylase ( P13051 ) activity has been hypothesized to influence the toxicity induced by TS inhibitors . In this study we created a human cell line expressing the Ugi protein inhibitor of P13051 family of UDGs , which reduces cellular P13051 activity by at least 45-fold . Genomic uracil incorporation was directly measured by mass spectrometry following treatment with TS inhibitors . Genomic uracil levels were increased over 4-fold following TS inhibition in the Ugi-expressing cells , but did not detectably increase in P13051 proficient cells . Despite the difference in genomic uracil levels , there was no difference in toxicity between the P13051 proficient and P13051 -inhibited cells to folate or nucleotide-based inhibitors of TS . Cell cycle analysis showed that P13051 proficient and P13051 -inhibited cells arrested in early S-phase and resumed replication progression during recovery from RTX treatment almost identically . The induction of gamma- P16104 was measured following TS inhibition as a measure of whether uracil excision promoted DNA double strand break formation during S-phase arrest . Although gamma- P16104 was detectable following TS inhibition , there was no difference between P13051 proficient and P13051 -inhibited cells . We therefore conclude that uracil excision initiated by P13051 does not adequately explain the toxicity caused by TS inhibition in this model . DB00104 is the favorable alternative for cisplatin resistance reversal of ovarian cancer in vitro and in nude mice in vivo . This study aimed to observe the effects of octreotide ( O75051 ) on cisplatin resistance reversal of cancer cells in vitro and in nude mice in vivo . MTT method and flow cytometry were used to investigate the effect of cisplatin , O75051 or the combination of these two compounds on the proliferation and apoptosis of SKOV3- O60220 cells . The size and weight of xenograft tumors from the nude mice model were measured . Real-time PCR was used to detect the mRNA expression of P30874 , P08183 , Q92887 , Q86UG4 -pi and P00533 in SKOV3/ O60220 cells following the different treatment . At the concentration of 2.5-20 g/ml , O75051 significantly reduced IC50 ( p < 0.05 ) and promoted apoptosis ( p < 0.05 ) of SKOV3- O60220 cells ' response to cisplatin . Unchanged expression was found in P30874 on the SKOV3/ O60220 cell in vitro after O75051 treatment , but increased expression in vivo ( p < 0.05 ) . O75051 increased Q86UG4 -pi expression ( p < 0.05 ) and reduced Q92887 and P00533 expression ( p < 0.05 ) in a dose-dependent manner . The similar results were obtained in mice in vivo experiment , except the reduced expression of Q86UG4 -pi . It is suggested that O75051 could inhibit ovarian cancer proliferation and promote apoptosis , via the cell surface P30874 , and reverse cisplatin resistance through inhibition of Q92887 , P00533 , and even Q86UG4 -pi expressions . Revelation of p53-independent function of Q13330 in DNA damage response via modulation of the P38936 P38936 -proliferating cell nuclear antigen pathway . Although metastasis-associated protein 1 ( Q13330 ) , a component of the nucleosome remodeling and deacetylase ( NuRD ) complex , is a DNA-damage response protein and regulates p53-dependent DNA repair , it remains unknown whether Q13330 also participates in p53-independent DNA damage response . Here , we provide evidence that Q13330 is a p53-independent transcriptional corepressor of P38936 ( P38936 ) , and the underlying mechanism involves recruitment of Q13330 -histone deacetylase 2 ( Q92769 ) complexes onto two selective regions of the P38936 ( P38936 ) promoter . Accordingly , Q13330 depletion , despite its effect on p53 down-regulation , superinduces P38936 ( P38936 ) , increases P38936 ( P38936 ) binding to proliferating cell nuclear antigen ( P12004 ) , and decreases the nuclear accumulation of P12004 in response to ionizing radiation . In support of a p53-independent role of Q13330 in DNA damage response , we further demonstrate that induced expression of Q13330 in p53-null cells inhibits P38936 ( P38936 ) promoter activity and P38936 ( P38936 ) binding to P12004 . Consequently , Q13330 expression in p53-null cells results in increased induction of gamma P16104 foci and DNA double strand break repair , and decreased DNA damage sensitivity following ionizing radiation treatment . These findings uncover a new target of Q13330 and the existence of an additional p53-independent role of Q13330 in DNA damage response , at least in part , by modulating the P38936 ( P38936 )- P12004 pathway , and thus , linking two previously unconnected NuRD complex and DNA-damage response pathways . Activation status of receptor tyrosine kinase downstream pathways in primary lung adenocarcinoma with reference of P01116 and P00533 mutations . The activation status of signal transduction pathways involving receptor tyrosine kinases and its association with P00533 or P01116 mutations have been widely studied using cancer cell lines , although it is still uncertain in primary tumors . To study the activation status of main components of growth factor-induced pathways , phosphorylated Akt ( pAkt ) , extracellular signal-regulated kinases 1 and 2 ( pERK ) and other downstream proteins were immunohistochemically examined using surgical samples of 193 primary lung adenocarcinomas . Also , thyroid transcription factor-1 ( Q15669 -1 ) expression and mutation status of P00533 and P01116 were examined . Advanced tumor stages ( p < 0.001 ) , negative Q15669 -1 expression ( p < 0.001 ) and Akt activation ( p=0.015 ) were independent and significant poor prognostic markers . Akt activation related to advanced stage ( p=0.021 ) , invasiveness ( p=0.004 ) , and not to mutations . Q15669 -1 expression associated with never-smoker ( p=0.013 ) , pre- or minimally invasiveness ( p < 0.001 ) and P00533 mutations ( p=0.017 ) as well as with pERK ( p=0.039 ) expression . P00533 mutations did not correlated with pAkt and pERK expression , which was different from the results based on cultured cells , while P01116 mutations were solely and significantly linked to P29323 activation ( p=0.009 ) . In lung adenocarcinoma , tumors with Q15669 -1 expression have distinct characteristics regarding mutations , signal protein activation and clinical issues . Moreover , this property was revealed to be important in outcome estimation at any tumor stage , whereas Akt activation is abnormally affected according to the tumor stage regardless of their cell origin . The signal proteins were differently related to mutation status from cultured cells . Molecular targeting therapy against promyelocytic leukemia protein using arsenic acids in experimental intracranial medulloblastoma . Our previous study using human Daoy medulloblastoma cells showed that the promyelocytic leukemia ( P29590 ) gene was significantly upregulated ( 2.5-fold ) in cells positive to prominin-1 antigen ( CD133 ) , a possible marker for cancer initiating cells . DB01169 ( As(2)O(3) ) is known to degrade P29590 protein and has been used for the treatment of patients with acute P29590 . In the present study , the effect of P29590 targeting therapy with As(2)O(3) and cytarabine ( DB00987 ) on Daoy medulloblastoma cell proliferation was investigated . Daoy cells were pretreated with As(2)O(3) for 6 weeks . The As(2)O(3)-pretreated Daoy cells were cultured in medium containing DB00987 and cell viability was examined . Next , the As(2)O(3)-pretreated Daoy cells were inoculated into the nude mouse brain and the effect of DB00987 on the tumor size was evaluated . A significant increase in chemosensitivity to DB00987 was observed in the As(2)O(3)-pretreated Daoy cells in both in vitro and in vivo conditions . P29590 and P24385 ( cyclin D1 ) protein expression of Daoy medulloblastoma cells was downregulated by As(2)O(3) treatment . P29590 has been proposed as a novel therapeutic target to eradicate quiescent leukemia-initiating cells , and P29590 -expressing CD133-positive cells are similarly a potential therapeutic target of treatment for medulloblastoma .