interacts_with DB06603?
Regulation of P40763 by histone deacetylase-3 in diffuse large B-cell lymphoma : implications for therapy . Diffuse large B-cell lymphoma ( DLBCL ) with an activated B-cell ( DB01048 ) gene-expression profile has been shown to have a poorer prognosis compared with tumors with a germinal center B-cell type . ABC cell lines have constitutive activation of P40763 ; however , the mechanisms regulating P40763 signaling in lymphoma are unknown . In studies of class-I histone deacetylase ( HDAC ) expression , we found overexpression of O15379 in phospho P40763 -positive DLBCL and the O15379 was found to be complexed with P40763 . Inhibition of HDAC activity by DB06603 ( LBH589 ) increased p300-mediated P40763 (Lys685) acetylation with increased nuclear export of P40763 to the cytoplasm . HDAC inhibition abolished P40763 (Tyr705) phosphorylation with minimal effect on P40763 (Ser727) and O60674 tyrosine activity . pSTAT3(Tyr705)-positive DLBCLs were more sensitive to HDAC inhibition with LBH589 compared with pSTAT3(Tyr705)-negative DLBCLs . This cytotoxicity was associated with downregulation of the direct P40763 target Mcl-1 . O15379 knockdown upregulated P40763 (Lys685) acetylation but prevented P40763 (Tyr705) phosphorylation and inhibited survival of pSTAT3-positive DLBCL cells . These studies provide the rationale for targeting P40763 -positive DLBCL tumors with HDAC inhibitors . Molecular mechanisms involving prostate cancer racial disparity . African American ( AA ) men with prostate cancer ( PCa ) have worse disease , with a higher incidence , younger age and more advanced disease at diagnosis , and a worse prognosis , compared to Caucasian ( CA ) men . In addition to socioeconomic factors and lifestyle differences , molecular alterations contribute to this discrepancy . In this review , we summarize molecular genetics research results interrelated with the biology of PCa racial disparity . Androgen and androgen receptor ( AR ) pathways have long been associated with prostate growth . Racial differences have also been found among variants of the genes of the enzymes involved in androgen biosynthesis and metabolism , such as P31213 , P05093 , and P08684 . The levels of expression and CAG repeat length of AR also show racial divergence and may be critical molecular alterations for racial disparity . Growth factors and their receptors , which promote cancer cell growth , are another potential cause of the disparity ; both P00533 and P29323 , two of the most studied receptors , show interethnic differences . Differences have also been found among genes regulating cell apoptosis , such as P10415 , which is increased in PCa in the AA population . Recent developments in genetics , proteomics , and genomics , among other molecular biotechnologies , will greatly aid the advancement of translational research on PCa racial disparity , hopefully culminating in the discovery of novel mechanisms of disease , in addition to prognostic markers and novel therapeutic approaches . DB01645 down-regulates androgen receptor by modulating Q9UBN7 -Hsp90 chaperone function . P10275 ( AR ) is a ligand-activated transcription factor belonging to the steroid hormone receptor family and is very important for the development and progression of prostate cancer . The soy isoflavone genistein has been shown previously to down-regulate AR in androgen-dependent prostate cancer cell lines such as LNCaP . However , the mechanism(s) by which AR is down-regulated by genistein is still not known fully . We show a new mechanism by which genistein inhibits AR protein levels . We show that genistein-treated LNCaP cells exhibit increased ubiquitination of AR , suggesting that AR protein is down-regulated via a proteasome-mediated pathway . AR is normally stabilized by the chaperone activity of the heat shock protein Hsp90 . The increased ubiquitination of AR after genistein treatment is attributed to decreased Hsp90 chaperone activity as assessed by its increased functionally inactive acetylated form . Consistent with this result , we find that Q9UBN7 , which is a Hsp90 deacetylase , is inhibited by the antiestrogenic activity of genistein . Hence , in this study , we elucidate a novel mechanism of AR down-regulation by genistein through inhibition of Q9UBN7 -Hsp90 cochaperone function required to stabilize AR protein . Our results suggest that genistein could be used as a potential chemopreventive agent for prostate cancers along with known inhibitors of Q9UBN7 and Hsp90 . Requirement and epigenetics reprogramming of Nrf2 in suppression of tumor promoter TPA-induced mouse skin cell transformation by sulforaphane . Nrf2 is a transcription factor that plays critical roles in regulating the expression of cellular defensive antioxidants and detoxification enzymes . However , the role of Nrf2 and Nrf2 's epigenetics reprogramming in skin tumor transformation is unknown . In this study , we investigated the inhibitory role and epigenetics of Nrf2 on tumor transformation induced by 12-O-tetradecanoylphorbol-13-acetate ( TPA ) in mouse skin epidermal JB6 ( JB6 P+ ) cells and the anticancer effect of sulforaphane ( SFN ) , an isothiocyanate found in cruciferous vegetables . After five days of treatment , SFN significantly inhibited TPA-induced JB6 cellular transformation and SFN enhanced the nuclear translocation of Nrf2 and increased the mRNA and protein levels of the Nrf2 target genes P09601 , P15559 and P22309 . Knockdown of Nrf2 attenuated the induction of Nrf2 , P09601 and P15559 by SFN , enhanced TPA-induced colony formation and dampened the inhibitory effect of SFN on TPA-induced JB6 transformation . Epigenetics investigation using bisulfite genomic sequencing showed that SFN decreased the methylation ratio of the first 15 CpGs of the Nrf2 gene promoter , which was corroborated by increased Nrf2 mRNA expression . Furthermore , SFN strongly reduced the protein expression of DNA methyltransferases ( P26358 , DNMT3a and DNMT3b ) . SFN also inhibited the total histone deacetylase ( HDAC ) activity and decreased the protein expression of Q13547 , Q92769 , O15379 and P56524 . Collectively , these results suggest that the anti-cancer effect of SFN against TPA-induced neoplastic transformation of mouse skin could involve the epigenetic reprogramming of anti-cancer genes such as Nrf2 , leading to the epigenetic reactivation of Nrf2 and the subsequent induction of downstream target genes involved in cellular protection . Genetic polymorphisms , the metabolism of estrogens and breast cancer : a review . Breast cancer is the most common female cancer and the second cause of cancer death in women . Despite recent breakthroughs , much of the etiology of this disease is unknown and the most important risk factor , i.e. , exposure to endogenous and exogenous estrogen throughout life can not explain the heterogeneity of prognosis nor clinical features of patients . Recently , many gene polymorphisms in the metabolism of breast cancer have been described as possible neoplasm etiologic factors . This review is an attempt to summarize the current knowledge about these polymorphisms and to determine new target genes for diagnosis and treatment of the disease . Polymorphisms in the genes P05093 , P11511 , P04798 , P05177 , Q16678 , P22309 , P50225 , 17-hydroxysteroid-dehydrogenase , P21964 , Q86UG4 , P03372 , and Q92731 are described . DB04365 -induced phosphorylated p53 and P38936 ( P38936 ) protein expression is dependent on Q13315 /ATR and phosphatidylinositol-3-kinase in clone-9 cells . Betel-quid use is associated with liver cancer whereas its constituent arecoline is cytotoxic , genotoxic , and induces p53-dependent P38936 ( P38936 ) protein expression in Clone-9 cells ( rat hepatocytes ) . The ataxia telangiectasia mutated ( Q13315 ) /rad3-related ( ATR ) -p53- P38936 ( P38936 ) and the phosphatidylinositol-3-kinase ( PI3K ) -mammalian target of rapamycin ( P42345 ) pathways are involved in the DNA damage response and the pathogenesis of cancers . Thus , we studied the role of Q13315 /ATR and PI3K in arecoline-induced p53 and P38936 ( P38936 ) protein expression in Clone-9 cells . We found that arecoline ( 0.5 mM ) activated the Q13315 /ATR kinase at 30 min . The arecoline-activated Q13315 /ATR substrate contained p-p53Ser15 . Moreover , arecoline only increased the levels of the p-p53Ser6 , p-p53Ser15 , and p-p53Ser392 phosphorylated p53 isoforms among the known isoforms . Q13315 shRNA attenuated arecoline-induced p-p53Ser15 and P38936 ( P38936 ) at 24 h . DB04365 ( 0.5 mM ) increased phosphorylation levels of p-AktSer473 and p-mTORSer2448 at 30-60 min . Dominant-negative PI3K plasmids attenuated arecoline-induced P38936 ( P38936 ) , but not p-p53Ser15 , at 24 h . DB00877 attenuated arecoline-induced phosphrylated p-p53Ser15 , but not P38936 ( P38936 ) , at 24 h . Q13315 shRNA , but not dominant-negative PI3K plasmids , attenuated arecoline-induced P38936 ( P38936 ) gene transcription . We conclude that arecoline activates the Q13315 /ATR-p53- P38936 ( P38936 ) and the PI3K/Akt- P42345 -p53 pathways in Clone-9 cells . DB04365 -induced phosphorylated p-p53Ser15 expression is dependent on Q13315 whereas arecoline-induced P38936 ( P38936 ) protein expression is dependent on Q13315 and PI3K . Moreover , P38936 ( P38936 ) gene is transcriptionally induced by arecoline-activated Q13315 . DB01197 attenuates matrix metalloproteinase-2 and -9 in monocrotaline-induced right ventricular hypertrophy in rats . Little is known about the influence of angiotensin converting enzyme ( P12821 ) inhibitors on matrix metalloproteinase ( MMP ) in right ventricular remodeling . We investigated the effect of captopril , an P12821 inhibitor , on P08253 and P14780 in monocrotaline-induced right ventricular hypertrophy . Six-week-old male Wistar rats were injected intraperitoneally with monocrotaline ( 60 mg/kg ) or saline . The rats were administrated captopril ( 30 mg/kg per day ) or a vehicle orally for 24 days from the day of monocrotaline injection . At day 25 , echocardiography was performed and hearts were excised . Expressions and activities of P08253 and P14780 were measured by Western blotting and by gelatin zymography , respectively . In monocrotaline-injected rats , right ventricular weight/tail length ratio increased significantly . Histological analysis revealed cardiomyocyte hypertrophy and fibrosis in right ventricular sections . Echocardiography showed right ventricular dysfunction compared with saline-injected rats . The right ventricular hypertrophy , fibrosis , and dysfunction were inhibited by captopril . However , captopril did not attenuate an increase in pulmonary artery pressure . P08253 and P14780 expressions and activities in right ventricles increased significantly in monocrotaline-injected rats and captopril inhibited them . These findings indicate that captopril attenuates the development of monocrotaline-induced right ventricular hypertrophy in association with inhibition of P08253 and P14780 in rats . Autocrine regulation of γ-irradiation-induced DNA damage response via extracellular nucleotides-mediated activation of Q15077 and Q9H244 receptors . A key component of the response to DNA damage caused by ionizing radiation is DNA repair . Release of extracellular nucleotides , such as DB00171 , from cells plays a role in signaling via P2 receptors . We show here that release of DB00171 , followed by activation of P2Y receptors , is involved in the response to γ-irradiation-induced DNA damage . Formation of phosphorylated histone variant P16104 ( γ P16104 ) foci , which are induced in nuclei by DNA damage and contribute to accumulation of DNA-repair factors , was increased at 1-3h after γ-ray irradiation ( 2.0Gy ) of human lung cancer A549 cells . Focus formation was suppressed by pre-treatment with the ecto-nucleotidase apyrase . Pre-treatment with ecto-nucleotidase inhibitor ARL67156 or post-treatment with DB00171 or UTP facilitated induction of γ P16104 , indicating that extracellular nucleotides play a role in induction of γ P16104 foci . Next , we examined the effect of P2 receptor inhibitors on activation of ataxia telangiectasia mutated ( Q13315 ; a protein kinase ) and accumulation of Q12888 ( a DNA repair factor ) , both of which are important for DNA repair , at DNA damage sites . Q15077 receptor antagonist MRS2578 , Q9H244 receptor antagonist clopidogrel , and Q99572 receptor antagonists A438079 and oxATP significantly inhibited these processes . Release of DB00171 was detected within 2.5min after irradiation , but was blocked by A438079 . Activation of Q13315 and accumulation of Q12888 were decreased in Q15077 or Q9H244 receptor-knockdown cells . We conclude that autocrine/paracrine signaling through Q99572 -dependent DB00171 release and activation of Q15077 and Q9H244 receptors serves to amplify the cellular response to DNA damage caused by γ-irradiation . Epigenetics , an early event in the modulation of gene expression by inositol hexaphosphate in ethylnitrosourea exposed mouse lungs . Mechanisms of anticancer effects of inositol hexaphosphate are not fully understood . Epigenetic changes are the early changes in tumorigenesis . DNA methyl transferases , methyl CpG binding proteins , methyl CpG DNA binding domain protein , and histone deacetylases are the major molecules involved in epigenetics . We have shown the effects of IP6 at the molecular level in mouse lungs before the tumor is developed . After 3 mo of ENU exposure , there was no tumor formation , but there was hyperplasia and lymphocytic infiltration in the lungs . Inflammation and DNA damage repair enzymes P35354 and P40692 appear to be upregulated , whereas tumor suppressor gene p16 was downregulated by ENU . On the other hand , ENU exposure more or less upregulated the epigenetic events such as the expressions of P26358 , MeCP2 , Q9UIS9 , and Q13547 . This alteration was reduced by IP6 administration . Results were supported by modulation of global DNA methylation and the modulation of promoter CpG methylation of p16 , P40692 , and P35354 genes . Hence , this study indicates the possible role of epigenetics at the early stage of tumor development and in the regulation of gene expression by IP6 before the onset of ENU-induced lung tumors . Increasing P16070 +/ P25063 (-) tumor stem cells , and upregulation of P35354 and Q9UBN7 , as major functions of P04626 in breast tumorigenesis . BACKGROUND : Cancer cells are believed to arise primarily from stem cells . P16070 +/ P25063 (-) have been identified as markers for human breast cancer stem cells . Although , P04626 is a well known breast cancer oncogene , the mechanisms of action of this gene are not completely understood . Previously , we have derived immortal ( M13SV1 ) , weakly tumorigenic ( M13SV1R2 ) and highly tumorigenic ( M13SV1R2N1 ) cell lines from a breast epithelial cell type with stem cell phenotypes after successive SV40 large T-antigen transfection , X-ray irradiation and ectopic expression of P04626 /C-erbB2/neu . Recently , we found that M13SV1R2 cells became non-tumorigenic after growing in a growth factor/hormone-deprived medium ( R2d cells ) . RESULTS : In this study , we developed M13SV1R2N1 under the same growth factor/hormone-deprived condition ( R2N1d cells ) . This provides an opportunity to analyze P04626 effect on gene expression associated with tumorigenesis by comparative study of R2d and R2N1d cells with homogeneous genetic background except P04626 expression . The results reveal distinct characters of R2N1d cells that can be ascribed to P04626 : 1 ) development of fast-growing tumors ; 2 ) high frequency of P16070 +/ P25063 (-) cells ( ~50 % for R2N1d vs. ~10 % for R2d ) ; 3 ) enhanced expression of P35354 , Q9UBN7 mediated , respectively , by MAPK and PI3K/Akt pathways , and many genes associated with inflammation , metastasis , and angiogenesis . Furthermore , P04626 expression can be down regulated in non-adhering R2N1d cells . These cells showed longer latent period and lower rate of tumor development compared with adhering cells . CONCLUSIONS : P04626 may induce breast cancer by increasing the frequency of tumor stem cells and upregulating the expression of P35354 and Q9UBN7 that play pivotal roles in tumor progression . Interferon-γ promotes vascular remodeling in human microvascular endothelial cells by upregulating endothelin ( ET ) -1 and transforming growth factor ( TGF ) β2 . Systemic sclerosis ( SSc ) is a complex disease characterized by vascular alterations , activation of the immune system and tissue fibrosis . Previous studies have implicated activation of the interferon pathways in the pathogenesis of SSc . The goal of this study was to determine whether interferon type I and/or type II could play a pathogenic role in SSc vasculopathy . Human dermal microvascular endothelial cells ( HDMVECs ) and fibroblasts were obtained from foreskins of healthy newborns . The RT Profiler PCR Array System was utilized to screen for EndoMT genes . Treatment with IFN-α or IFN-γ downregulated Fli1 and P33151 . In contrast , IFN-α and IFN-γ exerted opposite effects on the expression of α-SMA , P29279 , ET-1 , and TGFβ2 , with IFN-α downregulating and IFN-γ upregulating this set of genes . Blockade of TGFβ signaling normalized IFN-γ-mediated changes in Fli1 , P33151 , P29279 , and ET-1 levels , whereas upregulation of α-SMA and TGFβ2 was not affected . DB00559 treatment was more effective than TGFβ blockade in reversing the actions of IFN-γ , including downregulation of α-SMA and TGFβ2 , suggesting that activation of the ET-1 pathway plays a main role in the IFN-γ responses in HDMECs . IFN-γ induced expression of selected genes related to endothelial-to-mesenchymal transition ( EndoMT ) , including Snail1 , P02751 , P05121 , Q15672 , P40763 , P41220 , and components of the WNT pathway . The effect of IFN-γ on EndoMT was mediated via TGFβ2 and ET-1 signaling pathways . This study demonstrates distinct effects of IFN-α and IFN-γ on the biology of vascular endothelial cells . IFN-γ may contribute to abnormal vascular remodeling and fibrogenesis in SSc , partially via induction of EndoMT . Comparison of the cytotoxicity of cladribine and clofarabine when combined with fludarabine and busulfan in AML cells : Enhancement of cytotoxicity with epigenetic modulators . Clofarabine ( Clo ) , fludarabine ( Flu ) , and busulfan ( Bu ) combinations are efficacious in hematopoietic stem cell transplantation for myeloid leukemia . We sought to determine whether the more affordable drug cladribine ( Clad ) can provide a viable alternative to Clo , with or without DB06603 ( Pano ) and DB01262 ( P22760 ) . Both Clad+Flu+Bu and Clo+Flu+Bu combinations showed synergistic cytotoxicity in KBM3/Bu250(6) , HL60 , and OCI- Q13950 cell lines . Cell exposure to these drug combinations resulted in 60 % -80 % inhibition of proliferation ; activation of the Q13315 pathway ; increase in histone modifications ; decrease in O15379 , P56524 , Q9UQL6 and SirT7 proteins ; decrease in mitochondrial membrane potential ; activation of apoptosis and stress signaling pathways ; and downregulation of the AKT pathway . These drug combinations activated DNA-damage response and apoptosis in primary cell samples from AML patients . At lower concentrations of Clad/Clo , Flu , and Bu , inclusion of Pano and P22760 enhanced cell killing , increased histone modifications and DNA demethylation , and increased the levels of P16/INK4a , P15/INK4b and P21/Waf1/Cip1 proteins . The observed DNA demethylating activity of Clad and Clo may complement P22760 activity ; increase demethylation of the gene promoters for Q8N474 , Q9UBP4 , and Q9Y5W5 ; and cause degradation of β-catenin in cells exposed to Clad/Clo+Flu+Bu+ P22760 +Pano . The overlapping activities of Clad/Clo+Flu+Bu , Pano , and P22760 in DNA-damage formation and repair , histone modifications , DNA demethylation , and apoptosis may underlie their synergism . Our results provide a basis for supplanting Clo with Clad and for including epigenetic modifiers in the pre-hematopoietic stem cell transplantation conditioning regimen for myeloid leukemia patients . Expression of aberrant HLA- Q8TCY5 molecules is dependent on Q8TCY5 dosage and peptide supply . OBJECTIVES : Cellular expression of non-classical forms of human leukocyte antigen ( HLA ) - Q8TCY5 ( NC- Q8TCY5 ) may be involved in spondyloarthritis ( SpA ) pathogenesis . We used a novel Q8TCY5 -specific monoclonal antibody , Q9UBN7 , to ask if Q8TCY5 transgenic ( TG ) rat splenocytes express these NC- Q8TCY5 molecules . We also investigated whether Q8TCY5 -binding peptides could affect the expression and functional immune recognition of Q9UBN7 -reactive Q8TCY5 molecules . METHODS : Splenocytes from Q8TCY5 -TG , P33681 -TG and non-transgenic rats , and HLA- Q8TCY5 + cell lines were stained with monoclonal antibodies recognising classical ( ME-1 , HLA-ABC-m1 ) and non-classical ( Q9UBN7 , HC10 ) Q8TCY5 . Cells were further cultured in the presence of HLA- Q8TCY5 -binding peptides , or subjected to brief low pH treatment prior to mAb staining and/or immunoprecipitation or co-culture with P43630 -CD3ε-expressing Jurkat reporter cells . RESULTS : Q9UBN7 -reactive molecules were detected in the majority of adult Q8TCY5 -TG rat splenocyte cell subsets , increasing with age and concomitant increased Q8TCY5 expression . Q9UBN7 staining was inhibited by incubation with Q8TCY5 -binding peptides and induced by low pH treatment . Q9UBN7 staining correlated with P43630 -CD3ε-expressing Jurkat reporter cell activity . Thus , P60568 production was decreased when Q8TCY5 -expressing antigen-presenting cells were preincubated with Q8TCY5 -binding peptides , but increased following pretreatment with low pH buffer . CONCLUSIONS : Surface expression of Q9UBN7 -reactive Q8TCY5 molecules on Q8TCY5 -TG rat splenocytes is consistent with a pathogenic role for NC- Q8TCY5 in SpA . Interaction of NC- Q8TCY5 with innate immune receptors could be critical in SpA pathogenesis , and we show that this may be influenced by the availability and composition of the Q8TCY5 -binding peptide pool . Holocarboxylase synthetase interacts physically with nuclear receptor co-repressor , histone deacetylase 1 and a novel splicing variant of histone deacetylase 1 to repress repeats . P50747 ( holocarboxylase synthetase ) is a nuclear protein that catalyses the binding of biotin to distinct lysine residues in chromatin proteins . P50747 -dependent epigenetic marks are over-represented in repressed genomic loci , particularly in repeats . Evidence is mounting that P50747 is a member of a multi-protein gene repression complex , which determines its localization in chromatin . In the present study we tested the hypothesis that P50747 interacts physically with O75376 ( nuclear receptor co-repressor ) and Q13547 ( histone deacetylase 1 ) , thereby contributing toward the removal of H3K9ac ( Lys⁹-acetylated histone H3 ) gene activation marks and the repression of repeats . Physical interactions between P50747 and O75376 , Q13547 and a novel splicing variant of Q13547 were confirmed by co-immunoprecipitation , limited proteolysis and split luciferase complementation assays . When P50747 was overexpressed , the abundance of H3K9ac marks decreased by 50 % and 68 % in LTRs ( long terminal repeats ) 15 and 22 respectively in P29320 ( human embryonic kidney ) -293 cells compared with the controls . This loss of H3K9ac marks was linked with an 83 % decrease in mRNA coding for LTRs . Similar patterns were seen in pericentromeric alpha satellite repeats in chromosomes 1 and 4 . We conclude that interactions of P50747 with O75376 and HDACs contribute towards the transcriptional repression of repeats , presumably increasing genome stability . DB01032 reduces infection and inflammation in acute Pseudomonas aeruginosa pneumonia . The activation of inflammasome signaling mediates pathology of acute Pseudomonas aeruginosa pneumonia . This suggests that the inflammasome might represent a target to limit the pathological consequences of acute P. aeruginosa lung infection . Q96RD7 ( Px1 ) channels mediate the activation of caspase-1 and release of IL-1β induced by Q99572 receptor activation . The approved drug probenecid is an inhibitor of Px1 and DB00171 release . In this study , we demonstrate that probenecid reduces infection and inflammation in acute P. aeruginosa pneumonia . Treatment of mice prior to infection with P. aeruginosa resulted in an enhanced clearance of P. aeruginosa and reduced levels of inflammatory mediators , such as IL-1β . In addition , probenecid inhibited the release of inflammatory mediators in murine alveolar macrophages and human U937 cell-derived macrophages upon bacterial infection but not in human bronchial epithelial cells . Thus , Px1 blockade via probenecid treatment may be a therapeutic option in P. aeruginosa pneumonia by improving bacterial clearance and reducing negative consequences of inflammation . P42345 mediates human trophoblast invasion through regulation of matrix-remodeling enzymes and is associated with serine phosphorylation of P40763 . The intracellular signaling molecule mammalian target of rapamycin ( P42345 ) is essential for cell growth and proliferation . It is involved in mouse embryogenesis , murine trophoblast outgrowth and linked to tumor cell invasiveness . In order to assess the role of P42345 in human trophoblast invasion we analyzed the in vitro invasiveness of HTR-8/SVneo immortalized first-trimester trophoblast cells in conjunction with enzyme secretion upon P42345 inhibition and knockdown of P42345 protein expression . Additionally , we also tested the capability of P42345 to trigger signal transducer and activator of transcription ( P35610 ) -3 by its phosphorylation status . DB00877 inhibited P42345 kinase activity as demonstrated with a lower phosphorylation level of the P42345 substrate P08133 S6 kinase ( S6K ) . With the use of rapamycin and siRNA-mediated P42345 knockdown we could show that cell proliferation , invasion and secretion of matrix-metalloproteinases ( MMP ) -2 and -9 , urokinase-like plasminogen activator ( uPA ) and its major physiological uPA inhibitor ( P05121 ) -1 were inhibited . While tyrosine phosphorylation of P40763 was unaffected by P42345 inhibition and knockdown , serine phosphorylation was diminished . We conclude that P42345 signaling is one major mechanism in a tightly regulated network of intracellular signal pathways including the JAK/ P35610 system to regulate invasion in human trophoblast cells by secretion of enzymes that remodel the extra-cellular matrix ( Q13201 ) such as P08253 , -9 , uPA and P05121 . Dysregulation of P42345 may contribute to pregnancy-related pathologies caused through impaired trophoblast invasion . Combination therapy for hepatocellular carcinoma : additive preclinical efficacy of the HDAC inhibitor DB06603 with sorafenib . BACKGROUND & AIMS : Hepatocellular carcinoma ( HCC ) is a heterogeneous cancer in which sorafenib is the only approved systemic therapy . Histone deacetylases ( HDAC ) are commonly dysregulated in cancer and therefore represent promising targets for therapies , however their role in HCC pathogenesis is still unknown . We analyzed the expression of 11 HDACs in human HCCs and assessed the efficacy of the pan-HDAC inhibitor DB06603 alone and in combination with sorafenib in preclinical models of liver cancer . METHODS : Gene expression and copy number changes were analyzed in a cohort of 334 human HCCs , while the effects of DB06603 and sorafenib were evaluated in three liver cancer cell lines and a murine xenograft model . RESULTS : Aberrant HDAC expression was identified and validated in 91 and 243 HCCs , respectively . Upregulation of O15379 and Q9UQL6 mRNAs was significantly correlated with DNA copy number gains . Inhibiting HDACs with DB06603 led to strong anti-tumoral effects in vitro and vivo , enhanced by the addition of sorafenib . Cell viability and proliferation declined , while apoptosis and autophagy increased . DB06603 increased histone H3 and HSP90 acetylation , downregulated O15392 ( survivin ) and upregulated CDH1 . Combination therapy with DB06603 and sorafenib significantly decreased vessel density , and most significantly decreased tumor volume and increased survival in HCC xenografts . CONCLUSIONS : Aberrant expression of several HDACs and copy number gains of O15379 and Q9UQL6 occur in HCC . Treatment with DB06603 combined with sorafenib demonstrated the highest preclinical efficacy in HCC models , providing the rationale for clinical studies with this novel combination . Somatic mutations affect key pathways in lung adenocarcinoma . Determining the genetic basis of cancer requires comprehensive analyses of large collections of histopathologically well-classified primary tumours . Here we report the results of a collaborative study to discover somatic mutations in 188 human lung adenocarcinomas . DNA sequencing of 623 genes with known or potential relationships to cancer revealed more than 1,000 somatic mutations across the samples . Our analysis identified 26 genes that are mutated at significantly high frequencies and thus are probably involved in carcinogenesis . The frequently mutated genes include tyrosine kinases , among them the P00533 homologue Q15303 ; multiple ephrin receptor genes , notably P29320 ; vascular endothelial growth factor receptor P35968 ; and NTRK genes . These data provide evidence of somatic mutations in primary lung adenocarcinoma for several tumour suppressor genes involved in other cancers -- including P21359 , P25054 , P06400 and Q13315 -- and for sequence changes in P23468 as well as the frequently deleted gene Q9NZR2 . The observed mutational profiles correlate with clinical features , smoking status and DNA repair defects . These results are reinforced by data integration including single nucleotide polymorphism array and gene expression array . Our findings shed further light on several important signalling pathways involved in lung adenocarcinoma , and suggest new molecular targets for treatment . In vitro and in vivo evidence for a role of the Q99572 receptor in the release of P01584 in the murine brain . The P2X(7) receptor ( P2X(7)R ) is a purinoceptor expressed predominantly by cells of immune origin , including microglial cells . P2X(7)R has a role in the release of biologically active proinflammatory cytokines such as P01584 , P05231 and TNFalpha . Here we demonstrate that when incubated with lipopolysaccharide ( LPS ) , glial cells cultured from brain of P2X(7)R(-/-) mice produce less P01584 compared to glial cells from brains of wild-type mice . This is not the case for TNFalpha and P05231 . Our results indicate a selective effect of the P2X7R gene deletion on release of P01584 release but not of P05231 and TNFalpha . In addition , we confirm that only microglial cells produce IL-1beta , and this release is dependent on P2X(7)R and ABC1 transporter . Because P01584 is a key regulator of the brain cytokine network and P2X(7)R is an absolute requirement for P01584 release , we further investigated whether response of brain cytokines to LPS in vivo was altered in P2X(7)R(-/-) mice compared to wild-type mice . P01584 and TNFalpha mRNAs were less elevated in the brain of P2X(7)R(-/-) than in the brain of wild-type mice in response to systemic LPS . These results show that P2X7R plays a key role in the brain cytokine response to immune stimuli , which certainly applies also to cytokine-dependent alterations in brain functions including sickness behavior . R306465 is a novel potent inhibitor of class I histone deacetylases with broad-spectrum antitumoral activity against solid and haematological malignancies . R306465 is a novel hydroxamate-based histone deacetylase ( HDAC ) inhibitor with broad-spectrum antitumour activity against solid and haematological malignancies in preclinical models . R306465 was found to be a potent inhibitor of Q13547 and -8 ( class I ) in vitro . It rapidly induced histone 3 ( H3 ) acetylation and strongly upregulated expression of p21waf1,cip1 , a downstream component of Q13547 signalling , in A2780 ovarian carcinoma cells . R306465 showed class I HDAC isotype selectivity as evidenced by poor inhibition of Q9UBN7 ( class IIb ) confirmed by the absence of downregulation of Hsp90 chaperone c-raf protein expression and tubulin acetylation . This distinguished it from other HDAC inhibitors currently in clinical development that were either more potent towards Q9UBN7 ( e.g. vorinostat ) or had a broader HDAC inhibition spectrum ( e.g. DB06603 ) . R306465 potently inhibited cell proliferation of all main solid tumour indications , including ovarian , lung , colon , breast and prostate cancer cell lines , with IC50 values ranging from 30 to 300 nM . Haematological cell lines , including acute lymphoblastic leukaemia , acute myeloid leukaemia , chronic lymphoblastic leukaemia , chronic myeloid leukaemia , lymphoma and myeloma , were potently inhibited at a similar concentration range . R306465 induced apoptosis and inhibited angiogenesis in cell-based assays and had potent oral in vivo antitumoral activity in xenograft models . Once-daily oral administration of R306465 at well-tolerated doses inhibited the growth of A2780 ovarian , H460 lung and HCT116 colon carcinomas in immunodeficient mice . The high activity of R306465 in cell-based assays and in vivo after oral administration makes R306465 a promising novel antitumoral agent with potential applicability in a broad spectrum of human malignancies . Role of the androgen receptor axis in prostate cancer . P10275 ( AR ) is expressed in nearly all prostate cancers , including treatment-refractory disease . The role of this receptor in the molecular endocrinology of prostate cancer has become increasingly clear in recent years . The AR is now known to participate in tumor progression through 3 mechanisms : expression ( activation and upregulation of receptor activity ) , point mutations , and ligand-independent activation . With regard to the latter mechanism , interleukin-6 ( P05231 ) is among the most important nonsteroidal regulators of AR activity . In the absence of androgen , P05231 causes activation of AR that is approximately 50 % of the maximal activity induced by androgen . At low concentrations of androgen , P05231 and androgen synergistically activate AR . Nonsteroidal antiandrogens usually antagonize this activation , but they switch to an agonist effect in the presence of oncostatin M , an P05231 -related cytokine . The growth of parental LNCaP cells is initially inhibited by exposure to P05231 , but long-term treatment renders the cells resistant to such inhibition and confers a growth advantage . Both P05231 and oncostatin M stimulate AR activity , but only oncostatin M is associated with strong acquisition of the agonist properties of nonsteroidal antiandrogens . It is hoped that continuing research on AR expression and function in prostate cancer will pave the way for new therapeutic strategies . N-arachidonoyl-L-serine is neuroprotective after traumatic brain injury by reducing apoptosis . N-arachidonoyl-L-serine ( AraS ) is a brain component structurally related to the endocannabinoid family . We investigated the neuroprotective effects of AraS following closed head injury induced by weight drop onto the exposed fronto-parietal skull and the mechanisms involved . A single injection of AraS following injury led to a significant improvement in functional outcome , and to reduced edema and lesion volume compared with vehicle . Specific antagonists to CB2 receptors , transient receptor potential vanilloid 1 ( Q8NER1 ) or large conductance calcium-activated potassium ( BK ) channels reversed these effects . Specific binding assays did not indicate binding of AraS to the Q9Y2T6 cannabinoid receptor . N-arachidonoyl-L-serine blocked the attenuation in phosphorylated extracellular-signal-regulated kinase 1/2 ( P29323 ) levels and led to an increase in pAkt in both the ipsilateral and contralateral cortices . Increased levels of the prosurvival factor Bcl-xL were evident 24 hours after injury in AraS-treated mice , followed by a 30 % reduction in caspase-3 activity , measured 3 days after injury . Treatment with a CB2 antagonist , but not with a P21554 antagonist , reversed this effect . Our results suggest that administration of AraS leads to neuroprotection via P29323 and Akt phosphorylation and induction of their downstream antiapoptotic pathways . These protective effects are related mostly to indirect signaling via the CB2R and Q8NER1 channels but not through P21554 or Q9Y2T6 receptors . Cross-communication between histone H3 and H4 acetylation and Akt- P42345 signalling in prostate cancer cells . Molecular tumour targeting has significantly improved anti-cancer protocols . Still , the addition of molecular targeting to the treatment regime has not led to a curative breakthrough . Combined mammalian target of DB00877 ( P42345 ) and histone deacetylase ( HDAC ) inhibition has been shown not only to enhance anti-tumour potential , but also to prevent resistance development seen under mono-drug therapy . This investigation was designed to evaluate whether cross-communication exists between P42345 signalling and epigenetic events regulated by HDAC . DU-145 prostate cancer cells were treated with insulin-like growth factor ( IGF ) to activate the Akt- P42345 cascade or with the HDAC-inhibitor valproic acid ( DB00313 ) to induce histone H3 and H4 acetylation ( aH3 , aH4 ) . Subsequently , P42345 , Rictor , Q8N122 , p70s6k , Akt ( all : total and phosphorylated ) , H3 and H4 ( total and acetylated ) were analysed by western blotting . Both techniques revealed a link between P42345 and the epigenetic machinery . IGF activated P42345 , Rictor , Q8N122 , p70s6k and Akt , but also enhanced aH3 and aH4 . Inversely , IGFr blockade and knock-down blocked the Akt- P42345 axis , but simultaneously diminished aH3 and aH4 . DB00313 treatment up-regulated histone acetylation , but also activated P42345 -Akt signalling . Q13547 and 2 knock-down revealed that the interaction with the P42345 system is initiated by histone H3 acetylation . HDAC- P42345 communication , therefore , is apparent whereby tumour-promoting ( Akt/ P42345 (high) , aH3/aH4(low) ) and tumour-suppressing signals ( Akt/ P42345 (low) , aH3/aH4(high) ) are activated in parallel . Combined use of an HDAC- and P42345 inhibitor might then diminish pro-tumour effects triggered by the HDAC- ( Akt/ P42345 (high) ) or P42345 inhibitor ( aH3/aH4(low) ) alone . Antiangiogenic activity of the endocannabinoid anandamide : correlation to its tumor-suppressor efficacy . Endocannabinoids are now emerging as suppressors of key cell-signaling pathways involved in cancer cell growth , invasion , and metastasis . We have previously observed that the metabolically stable anandamide analog , 2-methyl-2'-F-anandamide ( DB00134 -F-AEA ) can inhibit the growth of thyroid cancer in vivo . Our hypothesis was that the anti-tumor effect observed could be at least in part ascribed to inhibition of neo-angiogenesis . Therefore , the aim of this study was to assess the anti-angiogenic activity of DB00134 -F-AEA , to investigate the molecular mechanisms underlying this effect and whether DB00134 -F-AEA could antagonize tumor-induced endothelial cell sprouting . We show that DB00134 -F-AEA inhibited P09038 -stimulated endothelial cell proliferation , in a dose-dependent manner , and also induced apoptosis , both effects reliant on cannabinoid P21554 receptor stimulation . Analyzing the signaling pathways implicated in angiogenesis , we observed that the P09038 -induced P29323 phosphorylation was antagonized by DB00134 -F-AEA , and we found that p38 MAPK was involved in DB00134 -F-AEA-induced apoptosis . Moreover , DB00134 -F-AEA was able to inhibit bi-dimensional capillary-like tube formation and activity of matrix metalloprotease P08253 , a major matrix degrading enzyme . Importantly , we demonstrated that DB00134 -F-AEA is also functional in vivo since it inhibited angiogenesis in the chick chorioallantoic neovascularization model . Finally , DB00134 -F-AEA inhibited tumor-induced angiogenesis in a three-dimensional model of endothelial and thyroid tumor cell ( KiMol ) spheroids co-cultures in different 3-D polymeric matrices that resemble tumor microenvironment and architecture . Thus , our results suggest that anandamide could be involved in the control of cancer growth targeting both tumor cell proliferation and the angiogenic stimulation of the vasculature . Histone deacetylase inhibitor treatment dramatically reduces cholesterol accumulation in Niemann-Pick type C1 mutant human fibroblasts . Niemann-Pick type C ( NPC ) disease is predominantly caused by mutations in the O15118 protein that affect intracellular cholesterol trafficking and cause accumulation of unesterified cholesterol and other lipids in lysosomal storage organelles . We report the use of a series of small molecule histone deacetylase ( HDAC ) inhibitors in tissue culture models of NPC human fibroblasts . Some HDAC inhibitors lead to a dramatic correction in the NPC phenotype in cells with either one or two copies of the O15118 (I1061T) mutation , and for several of the inhibitors , correction is associated with increased expression of O15118 protein . Increased O15118 (I1061T) protein levels may partially account for the correction of the phenotype , because this mutant can promote cholesterol efflux if it is delivered to late endosomes and lysosomes . The HDAC inhibitor treatment is ineffective in an P61916 mutant human fibroblast line . Analysis of the isoform selectivity of the compounds used implicates Q13547 and/or Q92769 as likely targets for the observed correction , although other HDACs may also play a role . LBH589 ( DB06603 ) is an orally available HDAC inhibitor that crosses the blood-brain barrier and is currently in phase III clinical trials for several types of cancer . It restores cholesterol homeostasis in cultured O15118 mutant fibroblasts to almost normal levels within 72 h when used at 40 nM . The findings that HDAC inhibitors can correct cholesterol storage defects in human O15118 mutant cells provide the potential basis for treatment options for NPC disease . Macroautophagy modulates cellular response to proteasome inhibitors in cancer therapy . Macroautophagy and the ubiquitin-proteasome system are two complementary pathways for protein degradation . The former degrades long-lived proteins and damaged organelles while the later degrades short-lived proteins . Recent findings indicate that suppression of the ubiquitin-proteasome system by proteasome inhibitors induces macroautophagy through multiple pathways , including ( 1 ) accumulation of ubiquitinated proteins and activation of Q9UBN7 ; ( 2 ) activation of the O75460 -JNK pathway ; ( 3 ) proteasomal stabilization of P18848 ; ( 4 ) inhibition of P42345 complex 1 signaling ; ( 5 ) reduced proteasomal degradation of LC3 . Induction of macroautophagy attenuates the antitumor effect of proteasome inhibitors in various types of cancer . These findings suggest that inhibition of macroautophagy may represent a novel strategy to enhance cellular sensitivity to proteasome inhibition . Involvement of the Q99572 purinergic receptor and c-Jun N-terminal and extracellular signal-regulated kinases in cyclooxygenase-2 and prostaglandin E2 induction by LL-37 . Periodontal disease is caused by microorganisms and host-derived inflammation involving increased cyclooxygenase-2 ( P35354 ) expression and prostaglandin E(2) ( PGE(2) ) production . We previously demonstrated that human β-defensin-3 induces P35354 and PGE(2) in human gingival fibroblasts ( HGFs ) . We , therefore , aimed to examine the inducible effects of LL-37 , the only cathelicidin expressed in humans , on P35354 expression and PGE(2) synthesis in HGFs and to elucidate the relevant signaling pathways . The P35354 expression was upregulated by LL-37 in dose- and time-dependent manners . Accordingly , the synthesis of PGE(2) in cell-free culture supernatants was raised by LL-37 ( p < 0.01 ) and blocked by NS-398 , a specific P35354 inhibitor ( p < 0.01 ) . P2X inhibitors and a neutralizing antibody against P2X(7) purinergic receptor significantly abrogated P35354 induction and PGE(2) production by LL-37 ( p < 0.01 ) . LL-37 upregulated P35354 expression and PGE(2) synthesis via activation of extracellular signal-regulated kinase ( P29323 ) and Q9BY77 c-Jun N-terminal kinase ( JNK ) , while interleukin-1β did so via nuclear factor-ĸB and all three mitogen-activated protein kinases . In summary , LL-37 can control arachidonic acid metabolism by induction of P35354 expression and PGE(2) synthesis via the P2X(7) receptor , P29323 , and Q9BY77 JNK . The pro-inflammatory effects of LL-37 may be essential for initiating oral mucosal inflammation in periodontal disease . Suppression of androgen receptor-mediated gene expression by a sequence-specific DNA-binding polyamide . P10275 ( AR ) is essential for the growth and progression of prostate cancer in both hormone-sensitive and hormone-refractory disease . A DNA-binding polyamide that targets the consensus androgen response element binds the prostate-specific antigen ( PSA ) promoter androgen response element , inhibits androgen-induced expression of PSA and several other AR-regulated genes in cultured prostate cancer cells , and reduces AR occupancy at the PSA promoter and enhancer . Down-regulation of PSA by this polyamide was comparable to that produced by the synthetic antiandrogen bicalutamide ( DB01128 ) at the same concentration . Genome-wide expression analysis reveals that a similar number of transcripts are affected by treatment with the polyamide and with bicalutamide . Direct inhibition of the AR-DNA interface by sequence-specific DNA binding small molecules could offer an alternative approach to antagonizing AR activity . The HDAC inhibitor LBH589 induces P29323 -dependent prometaphase arrest in prostate cancer via Q9UBN7 inactivation and down-regulation . Histone deacetylase inhibitors ( HDACIs ) have potent anti-cancer activity in a variety of cancer models . Understanding the molecular mechanisms involved in the therapeutic responsiveness of HDACI is needed before its clinical application . This study aimed to determine if a potent HDACI , LBH589 ( DB06603 ) , had differential therapeutic responsiveness towards LNCaP and PC-3 prostate cancer ( PCa ) cells . The former showed prometaphase arrest with subsequent apoptosis upon LBH589 treatment , while the latter was less sensitive and had late G2 arrest . The LBH589 treatment down-regulated Q9UBN7 and sustained P29323 activation , and contributed to prometaphase arrest . Mechanistically , LBH589 inhibited Q9UBN7 activity , caused its dissociation from protein phosphatase PP1α , and increased 14-3-3ζ acetylation . Acetylated 14-3-3ζ released its mask effect on serine 259 of c-Raf and serine 216 of Cdc25C subsequent to de-phosphorylation by PP1α , which contributed to P29323 activation . Enhanced P29323 activity by LBH589 further down-regulated Q9UBN7 protein levels and sustained P29323 activation by free-forward regulation . The sustained Cdc25C and P29323 activation resulted in early M-phase ( prometaphase ) arrest and subsequent apoptosis in the most sensitive LNCaP cells but not in PC-3 cells . This study provides pre-clinical evidence that Q9UBN7 may serve as a sensitive therapeutic target in the treatment of prostate cancer with HDACI LBH589 for clinical translation . This study also posits a novel mechanism of Q9UBN7 participation in regulating the c-Raf- P50391 - P29323 signaling pathway and contributing to M phase cell-cycle transition . Heterozygous mutation of ataxia-telangiectasia mutated gene aggravates hypercholesterolemia in apoE-deficient mice . Individuals with a heterozygous mutation at the ataxia-telangiectasia mutated gene ( Q13315 ) have been reported to be predisposed to ischemic heart disease . This report examined for the first time the effect of a heterozygous Q13315 mutation ( Q13315 (+)(/-) ) on plasma lipid levels and atherosclerosis intensity using Q13315 (+/-) , Q13315 (+)(/+) ( wild type ) , Q13315 (+)(/+)/ P01130 (-)(/-) ( low density lipoprotein receptor knockout ) , Q13315 (+)(/-)/ P01130 (-)(/-) , Q13315 (+)(/+)/ApoE(-)(/-) ( apolipoprotein E knockout ) , and Q13315 (+)(/-)/ApoE(-)(/-) mice . Our data demonstrated that the plasma cholesterol and triglyceride levels in Q13315 (+)(/-) and Q13315 (+)(/-)/ P01130 (-)(/-) mice were approximately the same as those in Q13315 (+)(/+) and Q13315 (+)(/+)/ P01130 (-)(/-) control mice , respectively . In contrast , the plasma cholesterol level was significantly higher in Q13315 (+)(/-)/ApoE(-)(/-) mice than in Q13315 (+)(/+)/ApoE(-)(/-) control mice . In addition , the Q13315 (+)(/-)/ApoE(-)(/-) mice showed higher plasma apoB-48 levels , slower clearance for plasma apoB-48-carrying lipoproteins , and more advanced atherosclerotic lesions in the aorta compared with the Q13315 (+)(/+)/ApoE(-)(/-) mice . These novel results suggest that the product of Q13315 is involved in an apoE-independent pathway for catabolism of apoB-48-carrying remnants ; therefore , superimposition of a heterozygous Q13315 mutation onto an ApoE deficiency background reduces the clearance of apoB-48-carrying lipoproteins from the blood circulation and promotes the formation of atherosclerosis . Histone deacetylase inhibitors as potential therapeutic approaches for chordoma : an immunohistochemical and functional analysis . Chordomas are rare malignancies of the axial skeleton . Therapy is mainly restricted to surgery . This study investigates histone deacetylase ( HDAC ) inhibitors as potential therapeutics for chordomas . Immunohistochemistry ( IHC ) was performed using the HDAC 1-6 antibodies on 50 chordoma samples ( 34 primary tumors , 16 recurrences ) from 44 patients ( 27 male , 17 female ) . Pan-HDAC-inhibitors DB02546 ( DB02546 ) , DB06603 ( LBH-589 ) , and Belinostat ( PXD101 ) were tested for their efficacy in the chordoma cell line MUG-Chor1 via Western blot , cell cycle analysis , caspase 3/7 activity ( MUG-Chor1 , UCh-1 ) , cleaved caspase-3 , and PARP cleavage . p-Values below 0.05 were considered significant . IHC was negative for Q13547 , positive for Q92769 in most ( n = 36 ; 72 % ) , and for HDACs 3-6 in all specimens available ( n = 43 ; 86 % ) . Q9UBN7 expression was strongest . DB02546 and LBH-589 , but not PXD101 caused a significant increase of G2/M phase cells and of cleaved caspase-3 ( p = 0.0003 , and p = 0.0014 after 72 h , respectively ) , and a peak of caspase 3/7 activity . PARP cleavage confirmed apoptosis . The presented chordoma series expressed HDACs 2-6 with strongest expression of Q9UBN7 . DB02546 and LBH-589 significantly increased apoptosis and changed cell cycle distribution in vitro . HDAC-inhibitors should be further evaluated as therapeutic options for chordoma . AM2389 , a high-affinity , in vivo potent P21554 -receptor-selective cannabinergic ligand as evidenced by drug discrimination in rats and hypothermia testing in mice . RATIONALE : The endocannabinoid signaling system ( ECS ) has been targeted for developing novel therapeutics since ECS dysfunction has been implicated in various pathologies . Current focus is on chemical modifications of the hexahydrocannabinol ( HHC ) nabilone ( DB00486 (®) ) . OBJECTIVE : To characterize the novel , high-affinity cannabinoid receptor 1 ( CB(1)R ) HHC-ligand AM2389 [ 9β-hydroxy-3-(1-hexyl-cyclobut-1-yl)-hexahydrocannabinol in two rodent pre-clinical assays . MATERIALS AND METHODS : CB(1)R mediation of AM2389-induced hypothermia in mice was evaluated with AM251 , a CB(1)R-selective antagonist/inverse agonist . Additionally , two groups of rats discriminated the full cannabinergic aminoalkylindole AM5983 ( 0.18 and 0.56 mg/kg ) from vehicle 20 min post-injection in a two-choice operant conditioning task motivated by 0.1 % saccharin/water . Generalization/substitution tests were conducted with AM2389 , AM5983 , and Δ(9)-tetrahydrocannabinol ( Δ(9)-THC ) . RESULTS : Δ(9)-THC (30 mg/kg)-induced hypothermia exhibited a faster onset and shorter duration of action compared with AM2389 ( 0.1 and 0.3 mg/kg ) . AM251 ( 3 and 10 mg/kg ) attenuated/blocked hypothermia induced by 0.3 mg/kg AM2389 . In drug discrimination , the order of potency was AM2389 > AM5983 > Δ(9)-THC with ED(50) values of 0.0025 , 0.0571 , and 0.2635 mg/kg , respectively , in the low-dose condition . The corresponding ED(50) values in the high-dose condition were 0.0069 , 0.1246 , and 0.8438 mg/kg , respectively . Onset of the effects of AM2389 was slow with a protracted time-course ; the functional , perceptual in vivo half-life was approximately 17 h . CONCLUSIONS : This potent cannabinergic HHC exhibited a slow onset of action with a protracted time-course . The AM2389 chemotype appears well suited for further drug development , and AM2389 currently is used to probe behavioral consequences of sustained ECS activation . DB06603 synergizes with bortezomib to induce endoplasmic reticulum stress and ubiquitinated protein accumulation in renal cancer cells . BACKGROUND : Inducing endoplasmic reticulum ( ER ) stress is a novel strategy used to treat malignancies . Inhibition of histone deacetylase ( HDAC ) 6 by the HDAC inhibitor DB06603 hinders the refolding of unfolded proteins by increasing the acetylation of heat shock protein 90 . We investigated whether combining DB06603 with the proteasome inhibitor bortezomib would kill cancer cells effectively by inhibiting the degradation of these unfolded proteins , thereby causing ubiquitinated proteins to accumulate and induce ER stress . METHODS : Caki-1 , ACHN , and 769-P cells were treated with DB06603 and/or bortezomib . Cell viability , clonogenicity , and induction of apoptosis were evaluated . The in vivo efficacy of the combination was evaluated using a murine subcutaneous xenograft model . The combination-induced ER stress and ubiquitinated protein accumulation were assessed . RESULTS : The combination of DB06603 and bortezomib induced apoptosis and inhibited renal cancer growth synergistically ( combination indexes < 1 ) . It also suppressed colony formation significantly ( p < 0.05 ) . In a murine subcutaneous tumor model , a 10-day treatment was well tolerated and inhibited tumor growth significantly ( p < 0.05 ) . Enhanced acetylation of the Q9UBN7 substrate alpha-tubulin was consistent with the suppression of Q9UBN7 activity by DB06603 , and the combination was shown to induce ER stress and ubiquitinated protein accumulation synergistically . CONCLUSIONS : DB06603 inhibits renal cancer growth by synergizing with bortezomib to induce ER stress and ubiquitinated protein accumulation . The current study provides a basis for testing the combination in patients with advanced renal cancer . DB01197 reduced plasminogen activator inhibitor activity in patients with acute myocardial infarction . Recent clinical trials have demonstrated that the administration of angiotensin-converting enzyme ( P12821 ) inhibitors to patients with myocardial infarction reduces the incidence of recurrent myocardial infarction . It has also been reported that an elevated level of plasminogen activator inhibitor ( P05121 ) appears to constitute a marker of the risk of recurrent coronary thrombosis . To determine whether the P12821 inhibitor captopril reduces plasma P05121 inhibitor activity , we measured changes in plasma P05121 activity ( IU/ml ) , tissue plasminogen activator ( t-PA ) antigen ( ng/ml ) , and serum P12821 activity ( IU/L ) in 14 survivors of myocardial infarction receiving captopril therapy ( 37.5 mg daily ) and compared them with the values in 15 placebo-treated patients chosen at random . Blood sampling was performed at 07.00 h . In the captopril-treated group , serum P12821 activity decreased significantly , from 14.0 +/- 0.8 to 11.5 +/- 1.2 IU/L 24 h after captopril therapy ( p < 0.01 ) , and those of P05121 activity and t-PA antigen also decreased significantly-from 11.9 +/- 2.8 to 5.5 +/- 2.2 IU/ml ( p < 0.02 ) and from 9.9 +/- 1.0 to 7.5 +/- 0.9 ng/ml ( p < 0.05 ) , respectively 48 h after captopril therapy . However , the levels of P12821 activity , P05121 activity , and t-PA antigen remained unchanged during the study period in the placebo group . Thus , our data indicate that the administration of captopril to patients with acute myocardial infarction may result in a reduced frequency of recurrent coronary thrombosis by increasing fibrinolytic capacity . Histone deacetylase inhibitors modulate the transcriptional regulation of guanylyl cyclase/natriuretic peptide receptor-a gene : interactive roles of modified histones , histone acetyltransferase , p300 , AND Sp1 . Atrial natriuretic peptide ( P01160 ) binds guanylyl cyclase-A/natriuretic peptide receptor-A ( P16066 /NPRA ) and produces the intracellular second messenger , cGMP , which regulates cardiovascular homeostasis . We sought to determine the function of histone deacetylases ( HDACs ) in regulating Npr1 ( coding for P16066 /NPRA ) gene transcription , using primary mouse mesangial cells treated with class-specific HDAC inhibitors ( HDACi ) . Trichostatin A , a pan inhibitor , and mocetinostat ( DB05651 ) , a class I HDAC inhibitor , significantly enhanced Npr1 promoter activity ( by 8- and 10-fold , respectively ) , mRNA levels ( 4- and 5.3-fold , respectively ) , and NPRA protein ( 2.7- and 3.5-fold , respectively ) . However , MC1568 ( class II HDAC inhibitor ) had no discernible effect . Overexpression of Q13547 and Q92769 significantly attenuated Npr1 promoter activity , whereas O15379 and Q9BY41 had no effect . HDACi-treated cultured cells in vitro and intact animals in vivo showed significantly reduced binding of Q13547 and -2 and increased accumulation of acetylated H3- P35527 /14 and H4-K12 at the Npr1 promoter . Deletional analyses of the Npr1 promoter along with ectopic overexpression and inhibition of Sp1 confirmed that HDACi-induced Npr1 gene transcription is accomplished by Sp1 activation . Furthermore , HDACi attenuated the interaction of Sp1 with Q13547 /2 and promoted Sp1 association with p300 and p300/ DB02527 -binding protein-associated factor ; it also promoted the recruitment of p300 and p300/ DB02527 -binding protein-associated factor to the Npr1 promoter . Our results demonstrate that trichostatin A and DB05651 enhanced Npr1 gene expression through inhibition of Q13547 /2 and increased both acetylation of histones ( H3- P35527 /14 , H4-K12 ) and Sp1 by p300 , and their recruitment to Npr1 promoter . Our findings define a novel epigenetic regulatory mechanism that governs Npr1 gene transcription . Astroglial P21554 cannabinoid receptors regulate leptin signaling in mouse brain astrocytes . Type-1 cannabinoid ( P21554 ) and leptin ( ObR ) receptors regulate metabolic and astroglial functions , but the potential links between the two systems in astrocytes were not investigated so far . Genetic and pharmacological manipulations of P21554 receptor expression and activity in cultured cortical and hypothalamic astrocytes demonstrated that cannabinoid signaling controls the levels of ObR expression . Lack of P21554 receptors also markedly impaired leptin-mediated activation of signal transducers and activators of transcription 3 and 5 ( P40763 and P42229 ) in astrocytes . In particular , P21554 deletion determined a basal overactivation of P42229 , thereby leading to the downregulation of ObR expression , and leptin failed to regulate P42229 -dependent glycogen storage in the absence of P21554 receptors . These results show that P21554 receptors directly interfere with leptin signaling and its ability to regulate glycogen storage , thereby representing a novel mechanism linking endocannabinoid and leptin signaling in the regulation of brain energy storage and neuronal functions . O15379 acts as a negative regulator of angiogenesis . Histone deacetylase-3 ( O15379 ) is involved in cellular proliferation , apoptosis and transcriptional repression . However , the role of O15379 in angiogenesis remains unknown . O15379 negatively regulated the expression of angiogenic factors , such as P15692 and plasminogen activator inhibitor-1 ( P05121 ) . O15379 showed binding to promoter sequences of P05121 . O15379 activity was necessary for the expression regulation of P05121 by O15379 . P15692 decreased the expression of O15379 , and the down-regulation of O15379 enhanced endothelial cell tube formation . O15379 negatively regulated tumor-induced angiogenic potential . We show the novel role of O15379 as a negative regulator of angiogenesis . Role of acetylation and extracellular location of heat shock protein 90alpha in tumor cell invasion . Heat shock protein ( hsp ) 90 is an DB00171 -dependent molecular chaperone that maintains the active conformation of client oncoproteins in cancer cells . An isoform , hsp90alpha , promotes extracellular maturation of matrix metalloproteinase ( MMP ) -2 , involved in tumor invasion and metastasis . Knockdown of histone deacetylase ( HDAC ) 6 , which deacetylates lysine residues in hsp90 , induces reversible hyperacetylation and attenuates DB00171 binding and chaperone function of hsp90 . Here , using mass spectrometry , we identified seven lysine residues in hsp90alpha that are hyperacetylated after treatment of eukaryotic cells with a pan-HDAC inhibitor that also inhibits Q9UBN7 . Depending on the specific lysine residue in the middle domain involved , although acetylation affects DB00171 , cochaperone , and client protein binding to hsp90alpha , acetylation of all seven lysines increased the binding of hsp90alpha to 17-allyl-amino-demethoxy geldanamycin . Notably , after treatment with the pan-HDAC inhibitor DB06603 ( LBH589 ) , the extracellular hsp90alpha was hyperacetylated and it bound to P08253 , which was associated with increased in vitro tumor cell invasiveness . Treatment with antiacetylated hsp90alpha antibody inhibited in vitro invasion by tumor cells . Thus , reversible hyperacetylation modulates the intracellular and extracellular chaperone function of hsp90 , and targeting extracellular hyperacetylated hsp90alpha may undermine tumor invasion and metastasis . DB00175 -induced changes in receptor-mediated metabolism of low density lipoprotein in guinea pigs . The effect of pravastatin , an inhibitor of P04035 , on the metabolism of human low density lipoprotein ( LDL ) was examined in guinea pigs . DB00175 treatment significantly reduced plasma levels of total cholesterol and LDL-cholesterol by 15.6 mg/dl ( 38.8 % ) and 12.7 mg/dl ( 42.9 % ) , respectively . We investigated the metabolism of LDL in pravastatin-treated and untreated guinea pigs using the simultaneous intravenous injection of 131I-labeled LDL and 125I-labeled , galactose-treated LDL to quantify the P01130 pathway . DB00175 increased the fractional catabolic rate ( FCR ) of the P01130 -dependent pathway . The treatment with pravastatin did not alter the FCR of the P01130 -independent pathway . The FCR of the P01130 -dependent pathway was higher for LDL isolated from pravastatin-treated subjects than for LDL isolated from control subjects . These findings suggest that pravastatin mainly reduced plasma cholesterol levels by accelerated FCR of the P01130 -mediated pathway . Inducible raptor and rictor knockout mouse embryonic fibroblasts . The mammalian Target of DB00877 ( P42345 ) kinase functions within two structurally and functionally distinct multiprotein complexes termed P42345 complex 1 ( mTORC1 ) and mTORC2 . The immunosuppressant and anticancer drug rapamycin is commonly used in basic research as a tool to study P42345 signaling . However , rapamycin inhibits only , and only incompletely , mTORC1 , and no mTORC2-specific inhibitor is available . Hence , a full understanding of P42345 signaling in vivo , including the function of both complexes , requires genetic inhibition in addition to pharmacological inhibition . Taking advantage of the Cre/LoxP system , we generated inducible knockout mouse embryonic fibroblasts ( MEFs ) deficient for either the mTORC1-specific component raptor ( iRapKO ) or the mTORC2-specific component rictor ( iRicKO ) . Inducibility of the knockout was important because P42345 complex components are essential . Induction of either raptor or rictor knockout eliminated raptor or rictor expression , respectively , and impaired the corresponding P42345 signaling branch . The described knockout MEFs are a valuable tool to study the full function of the two P42345 complexes individually . P10275 signaling : mechanism of interleukin-6 inhibition . Nonsteroidal signaling via the androgen receptor ( AR ) plays an im-portant role in hormone-refractory prostate cancer . Previously , we have reported that the pleiotropic cytokine , interleukin ( IL ) -6 , inhibited dihydrotestosterone-mediated expression of prostate-specific antigen in LNCaP cells ( Jia et al. , Mol Can Res 2003;1:385-92 ) . In the present study , we explored the mechanisms involved in this inhibition and considered possible effects on AR nuclear translocation , recruitment of transcription cofactors , and the signaling pathways that may mediate this inhibitory effect . P05231 neither induced nuclear localization of the AR nor inhibited dihydrotestosterone-induced nuclear translocation of the receptor . P05231 did not affect AR or P52701 coactivator recruitment to the transcription initiation complex on the prostate-specific antigen enhancer and promoter . Moreover , it did not lead to the recruitment of the corepressor silencing mediator of retinoic acid and thyroid hormone receptor ( Q9Y618 ) or histone deacetylase 1 ( Q13547 ) at the same sites . P05231 did , however , prevent the recruitment of the secondary coactivator , p300 , to the complex and partially inhibited histone H3 acetylation at the same loci . Furthermore , inhibition by P05231 was not mediated by the mitogen-activated protein kinase or the Akt pathways and was partially abrogated by signal transducers and activators of transcription-3 knock-down using small interfering RNA . Our results show that P05231 modulates androgen action through the differential recruitment of cofactors to target genes . These findings may account for the pleiotropic actions of P05231 in malignant prostate cells . Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 and RFP-induced hepatic oxidative stress in mice . Wild type ( MT+/+ ) and MT-null ( MT-/- ) mice were treated intragastrically with DB00951 ( 150 mg/kg ) , RFP ( 300 mg/kg ) , or the combination ( 150 mg/kg DB00951 +300 mg/kg RFP ) for 21 days . The results showed that MT-/- mice were more sensitive than MT+/+ mice to DB00951 and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 increased the protein expression of hepatic P05181 and DB00951 /RFP ( alone or in combination ) decreased the expression of hepatic P05177 . These findings clearly demonstrate that basal MT provides protection against DB00951 and RFP-induced toxicity in hepatocytes . The P05181 and P05177 were involved in the pathogenesis of DB00951 and RFP-induced hepatotoxicity . Ellagic acid inhibits P15692 / P35968 , PI3K/Akt and MAPK signaling cascades in the hamster cheek pouch carcinogenesis model . BACKGROUND : Blocking vascular endothelial growth factor ( P15692 ) mediated tumor angiogenesis by phytochemicals has emerged as an attractive strategy for cancer prevention and therapy . METHODS : We investigated the anti-angiogenic effects of ellagic acid in a hamster model of oral oncogenesis by examining the transcript and protein expression of hypoxia-inducible factor-1alpha ( HIF-1α ) , P15692 , P35968 , and the members of the PI3K/Akt and MAPK signaling cascades . Molecular docking studies and cell culture experiments with the endothelial cell line ECV304 were performed to delineate the mechanism by which ellagic acid regulates P15692 signaling . RESULTS : We found that ellagic acid significantly inhibits HIF-1α-induced P15692 / P35968 signalling in the hamster buccal pouch by abrogating PI3K/Akt and MAPK signaling via downregulation of PI3K , PDK-1 , p-Akt(ser473) , P42345 , p- P29323 , and p-JNK . Ellagic acid was also found to reduce the expression of histone deacetylases that could inhibit neovascularization . Analysis of the mechanism revealed that ellagic acid inhibits hypoxia-induced angiogenesis via suppression of HDAC-6 in ECV304 cells . Furthermore , knockdown of endogenous Q9UBN7 via small interfering RNA abrogated hypoxia-induced expression of HIF-1α and P15692 and blocked Akt activation . Molecular docking studies confirmed interaction of ellagic acid with upstream kinases that regulate angiogenic signaling . CONCLUSIONS : Taken together , these findings demonstrate that the anti-angiogenic activity of ellagic acid may be mediated by abrogation of hypoxia driven PI3K/Akt/ P42345 , MAPK and P15692 / P35968 signaling pathways involving suppression of Q9UBN7 and HIF-1α responses . GENERAL SIGNIFICANCE : Ellagic acid offers promise as a lead compound for anticancer therapeutics by virtue of its ability to inhibit key oncogenic signaling cascades and HDACs . Role of phospholipase D2 in the agonist-induced and constitutive endocytosis of G-protein coupled receptors . We have recently shown that the mu-opioid receptor [ P35372 , also termed mu-opioid peptide ( MOP ) receptor ] is associated with the phospholipase D2 ( O14939 ) , a phospholipid-specific phosphodiesterase located in the plasma membrane . We further demonstrated that , in human embryonic kidney ( P29320 ) 293 cells co-expressing P35372 and O14939 , treatment with ( D-Ala2 , Me Phe4 , Glyol5 ) enkephalin ( DAMGO ) led to an increase in O14939 activity and an induction of receptor endocytosis , whereas morphine , which does not induce opioid receptor endocytosis , failed to activate O14939 . In contrast , a C-terminal splice variant of the mu-opioid receptor ( MOR1D , also termed MOP(1D) ) exhibited robust endocytosis in response to both DAMGO and morphine treatment . We report here that MOR1D also mediates an agonist-independent ( constitutive ) O14939 -activation facilitating agonist-induced and constitutive receptor endocytosis . Inhibition of O14939 activity by over-expression of a dominant negative O14939 ( nPLD2 ) blocked the constitutive O14939 activation and impaired the endocytosis of MOR1D receptors . Moreover , we provide evidence that the endocytotic trafficking of the delta-opioid receptor [ Q8IXH6 , also termed delta-opioid peptide ( DOP ) receptor ] and cannabinoid receptor isoform 1 ( P21554 ) is also mediated by a O14939 -dependent pathway . These data indicate the generally important role for O14939 in the regulation of agonist-dependent and agonist-independent G protein-coupled receptor ( GPCR ) endocytosis . Statins control oxidized LDL-mediated histone modifications and gene expression in cultured human endothelial cells . OBJECTIVE : Activation of the endothelium by oxidized low-density lipoprotein ( oxLDL ) has been implicated in the development of atherosclerosis . Histone modifications impact on the transcriptional activity state of genes . We tested the hypothesis that oxLDL-induced inflammatory gene expression is regulated by histone modifications and experienced the effect of statins on these alterations . METHODS AND RESULTS : OxLDL-related interleukin-8 ( P10145 ) and monocyte-chemoattractant protein-1 ( P13500 ) secretion in endothelial cells was reduced by statins but enhanced by histone deacetylase inhibitors . OxLDL induced lectin-like oxidized P01130 -1 ( P78380 ) and extracellular regulated kinases ( P27361 /2 ) -dependent acetylation of histone H3 and H4 as well as phosphorylation of histone H3 , both globally and on the promoters of il8 and mcp1 . Pretreatment of oxLDL-exposed cells with statins reduced the above mentioned histone modification , as well as recruitment of CREB binding protein ( CBP ) 300 , NF-kappaB , and of RNA polymerase II but prevented loss of binding of histone deacetylase ( HDAC ) -1 and -2 at the il8 and mcp1 gene promoters . OxLDL reduced Q13547 and 2 expression , and statins partly restored global HDAC-activity . Statin-related effects were reverted with mevalonate . In situ experiments indicated decreased expression of Q92769 in endothelial cells in atherosclerotic plaques of human coronary arteries . CONCLUSIONS : Histone modifications seem to play an important role in atherosclerosis . Interactions of alpha- , beta- , and theta-defensins with influenza A virus and surfactant protein D . We have reported that the alpha-defensins human neutrophil peptides ( HNP ) -1 and HNP-2 neutralize and aggregate influenza A virus ( IAV ) and promote uptake of IAV by neutrophils . These alpha-defensins were also shown to bind to surfactant protein ( SP ) -D and reduce its antiviral activity . In this study , we examined retrocyclin (RC)1 and RC2 , humanized versions of the antiviral theta-defensins found in the leukocytes of certain nonhuman primates . Q9HC52 was just as effective as P59665 -3 in neutralizing IAV , and RC2 and RC101 ( an analog of Q9HC52 ) were more effective . In contrast , human beta-defensins ( HBDs ) showed less neutralizing activity . Human defensins 5 and 6 ( mainly produced by intestinal Paneth cells ) had viral neutralizing activity similar to P59665 -3 . Like P59665 -3 , RCs induced viral aggregation and promoted the uptake of IAV by neutrophils . We used surface plasmon resonance to evaluate binding of defensins to P35247 . HBDs , Q9UBN7 , and P12838 bound minimally to P35247 . P59665 -3 and RCs bound P35247 with high affinity ; however , unlike P59665 and HNP-2 , RCs did not inhibit P35247 antiviral activity . HBDs also did not inhibit antiviral activity of P35247 . Given their strong neutralizing activity and compatibility with P35247 , RCs may provide attractive prototypes for designing therapeutics that can prevent or treat respiratory infections caused by IAV . [ The effect of bunazosin vs captopril on hemodynamic and neurohumoral parameters in patients with congestive heart failure ] . The hemodynamic parameters ( right atrial pressure , mean pulmonary artery pressure , pulmonary capillary wedge pressure , cardiac index , heart rate , blood pressure ) and neurohumoral responses ( alpha- P01160 , plasma renin activity , aldosterone , angiotensin II ) of DB01197 , oral P12821 inhibitor , and Bunazosin , oral alpha 1-blocker , were investigated in 28 patients with congestive heart failure at rest and after exercise . These data were analysed in both acute and chronic phases . 1 ) Acute effect . DB01197 produced significant improvement of neurohumoral factors at rest and also after exercise . Bunazosin reduced alpha- P01160 , but other neurohumoral factors did not change . Bunazosin produced significant hemodynamic improvement both at rest and after exercise . 2 ) Chronic effect . DB01197 produced significant hemodynamic improvement both at rest and after exercise . Improvement of neurohumoral factors in acute phase was also preserved at chronic phase . On Bunazosin , improvement of hemodynamics at acute phase was also preserved at chronic phase without deterioration of neurohumoral factors . Loss of Androgen-Regulated MicroRNA 1 Activates P12931 and Promotes Prostate Cancer Bone Metastasis . Bone metastasis is the hallmark of progressive and castration-resistant prostate cancers . MicroRNA 1 ( miR-1 ) levels are decreased in clinical samples of primary prostate cancer and further reduced in metastases . P12931 has been implicated as a critical factor in bone metastasis , and here we show that P12931 is a direct target of miR-1 . In prostate cancer patient samples , miR-1 levels are inversely correlated with P12931 expression and a P12931 -dependent gene signature . Ectopic miR-1 expression inhibited extracellular signal-regulated kinase ( P29323 ) signaling and bone metastasis in a xenograft model . In contrast , P12931 overexpression was sufficient to reconstitute bone metastasis and P29323 signaling in cells expressing high levels of miR-1 . P10275 ( AR ) activity , defined by an AR output signature , is low in a portion of castration-resistant prostate cancer . We show that AR binds to the miR-1-2 regulatory region and regulates miR-1 transcription . Patients with low miR-1 levels displayed correlated low canonical AR gene signatures . Our data support the existence of an AR-miR-1- P12931 regulatory network . We propose that loss of miR-1 is one mechanistic link between low canonical AR output and P12931 -promoted metastatic phenotypes . Mammalian Q99572 receptor pharmacology : comparison of recombinant mouse , rat and human Q99572 receptors . BACKGROUND AND PURPOSE : Acute activation of Q99572 receptors rapidly opens a non-selective cation channel . Sustained Q99572 receptor activation leads to the formation of cytolytic pores , mediated by downstream recruitment of hemichannels to the cell surface . Species- and single-nucleotide polymorphism-mediated differences in Q99572 receptor activation have been reported that complicate understanding of the physiological role of Q99572 receptors . Studies were conducted to determine pharmacological differences between human , rat and mouse Q99572 receptors . EXPERIMENTAL APPROACH : Receptor-mediated changes in calcium influx and Yo-Pro uptake were compared between recombinant mouse , rat and human Q99572 receptors . For mouse Q99572 receptors , wild-type ( BALB/c ) and a reported loss of function ( C57BL/6 ) Q99572 receptor were also compared . KEY RESULTS : BzATP [ 2,3-O-(4-benzoylbenzoyl)- DB00171 ] was more potent than DB00171 in stimulating calcium influx and Yo-Pro uptake at rat , human , BALB/c and C57BL/6 mouse Q99572 receptors . Two selective Q99572 receptor antagonists , A-740003 and A-438079 , potently blocked Q99572 receptor activation across mammalian species . Several reported P51575 receptor antagonists [ e.g. P59665 2159 ( 4- [ ( 4-formyl-5-hydroxy-6-methyl-3- [ (phosphonooxy)methyl } -2-pyridinyl ) azo ] -benzoic acid ) , PPNDS and NF279 ] blocked Q99572 receptors . NF279 fully blocked human Q99572 receptors , but only partially blocked BALB/c Q99572 receptors and was inactive at C57BL/6 Q99572 receptors . CONCLUSIONS AND IMPLICATIONS : These data provide new insights into Q99572 receptor antagonist pharmacology across mammalian species . Q99572 receptor pharmacology in a widely used knockout background mouse strain ( C57BL/6 ) was similar to wild-type mouse Q99572 receptors . Several structurally novel , selective and competitive Q99572 receptor antagonists show less species differences compared with earlier non-selective antagonists . A transcriptional repressor co-regulatory network governing androgen response in prostate cancers . Transcriptional corepressors are frequently aberrantly over-expressed in prostate cancers . However , their crosstalk with the P10275 ( AR ) , a key player in prostate cancer development , is unclear . Using ChIP-Seq , we generated extensive global binding maps of AR , ERG , and commonly over-expressed transcriptional corepressors including Q13547 , Q92769 , O15379 , and Q15910 in prostate cancer cells . Surprisingly , our results revealed that ERG , HDACs , and Q15910 are directly involved in androgen-regulated transcription and wired into an AR centric transcriptional network via a spectrum of distal enhancers and/or proximal promoters . Moreover , we showed that similar to ERG , these corepressors function to mediate repression of AR-induced transcription including cytoskeletal genes that promote epithelial differentiation and inhibit metastasis . Specifically , we demonstrated that the direct suppression of P18206 expression by ERG , Q15910 , and HDACs leads to enhanced invasiveness of prostate cancer cells . Taken together , our results highlight a novel mechanism by which , ERG working together with oncogenic corepressors including HDACs and the polycomb protein , Q15910 , could impede epithelial differentiation and contribute to prostate cancer progression , through directly modulating the transcriptional output of AR . Q9UBC3 interacts with O60264 chromatin remodeling enzyme , HDACs 1 and 2 , and components of the histone methylation system . The non-random pattern of genome-wide DNA methylation in mammalian cells is established and maintained by DNA methyltransferases P26358 , 3A , and 3B . De novo DNA methyltransferase Q9UBC3 is critical for embryonic development and is mutated in ICF syndrome . Despite its importance in normal cellular functioning , little is known about how Q9UBC3 operates in the context of chromatin . Here we demonstrate that Q9UBC3 associates with four chromatin-associated enzymatic activities common to transcriptionally repressed , heterochromatic regions of the genome : DNA methyltransferase , histone deacetylase , ATPase , and histone methylase activities . By immunoprecipitation and Q86UG4 pull-down , we show that Q9UBC3 interacts with Q13547 , Q92769 , P59665 proteins , Suv39h1 , and the DB00171 -dependent chromatin remodeling enzyme O60264 . Endogenous O60264 is also associated with DNA methyltransferase activity . These proteins co-localize extensively with Q9UBC3 in heterochromatic regions . Our results therefore link Q9UBC3 to three other components of the epigenetic machinery and provide important insights into how DNA methylation patterns may be established within the chromatin environment . Deciphering the molecular and biologic processes that mediate histone deacetylase inhibitor-induced thrombocytopenia . Histone deacetylase inhibitor ( HDACI ) -induced thrombocytopenia ( DB01520 ) is a major dose-limiting toxicity of this new class of drugs . Using preclinical models to study the molecular and biologic events that underpin this effect of HDACI , we found that C57BL/6 mice treated with both the Q13547 /2-selective HDACI romidepsin and the pan-HDACI DB06603 developed significant DB01520 . HDACI-induced DB01520 was not due to myelosuppression or reduced platelet lifespan , but to decreased platelet release from megakaryocytes . Cultured primary murine megakaryocytes showed reductions in proplatelet extensions after HDACI exposure and a dose-dependent increase in the phosphorylation of myosin light chain 2 ( MLC2 ) . Phosphorylation of MLC to phospho-MLC ( pMLC ) and subsequent proplatelet formation in megakaryocytes is regulated by the Rho-GTPase proteins Rac1 , P60953 , and RhoA . Primary mouse megakaryocytes and the human megakaryoblastic cell line Meg-01 showed reductions in Rac1 , P60953 , and RhoA protein levels after treatment with HDACIs . We were able to overcome HDACI-induced DB01520 by administering the mouse-specific thrombopoietin ( P07202 ) mimetic AMP-4 , which improved platelet numbers to levels similar to untreated controls . Our report provides the first detailed account of the molecular and biologic processes involved in HDACI-mediated DB01520 . Moreover , our preclinical studies provide evidence that dose-limiting DB01520 induced by HDACIs may be circumvented using a P07202 mimetic . Gossypin up-regulates P01130 through activation of P29323 pathway : a signaling mechanism for the hypocholesterolemic effect . Hypercholesterolemia is one of the major risk factors for the development of cardiovascular disease . This study aims to elucidate the effect of gossypin on cholesterol metabolism in HepG2 cells . Results indicated that gossypin significantly reduced the total cholesterol concentration in a dose-dependent manner . There was a time- and dose-dependent increase in the expression of low-density lipoprotein receptor ( P01130 ) protein . However , 3-hydroxy-3-methylglutaryl coenzyme A ( HMG- DB01992 ) reductase , the rate-limiting enzyme in cholesterol synthesis , was not affected by gossypin . Moreover , gossypin had no effect on nuclear sterol regulatory element binding proteins ( SREBP ) -2 abundance . The activity of gossypin on P01130 expression was inhibited by the extracellular signal-regulated kinase ( P29323 ) inhibitor PD98059 . Western blotting analysis revealed that gossypin treatment dose- and time-dependently increased P29323 activation and preceded the up-regulation of P01130 expression . Collectively , these new findings identify gossypin as a new hypocholesterolemic agent that up-regulates P01130 expression independent of Q12772 but is dependent on P29323 activation . Determination of the class and isoform selectivity of small-molecule histone deacetylase inhibitors . The human HDAC ( histone deacetylase ) family , a well-validated anticancer target , plays a key role in the control of gene expression through regulation of transcription . While HDACs can be subdivided into three main classes , the class I , class II and class III HDACs ( sirtuins ) , it is presently unclear whether inhibiting multiple HDACs using pan-HDAC inhibitors , or targeting specific isoforms that show aberrant levels in tumours , will prove more effective as an anticancer strategy in the clinic . To address the above issues , we have tested a number of clinically relevant HDACis ( HDAC inhibitors ) against a panel of rhHDAC ( recombinant human HDAC ) isoforms . Eight rhHDACs were expressed using a baculoviral system , and a Fluor de Lystrade mark ( Biomol International ) HDAC assay was optimized for each purified isoform . The potency and selectivity of ten HDACs on class I isoforms ( rhHDAC1 , rhHDAC2 , rhHDAC3 and rhHDAC8 ) and class II HDAC isoforms ( rhHDAC4 , rhHDAC6 , rhHDAC7 and rhHDAC9 ) was determined . MS-275 was Q13547 -selective , DB05651 was Q13547 - and Q92769 -selective , apicidin was Q92769 - and O15379 -selective and valproic acid was a specific inhibitor of class I HDACs . The hydroxamic acid-derived compounds ( trichostatin A , DB00238 -LAQ824 , DB06603 , ITF2357 , vorinostat and belinostat ) were potent pan-HDAC inhibitors . The growth-inhibitory effect of the HDACis on HeLa cells showed that both pan-HDAC and class-I-specific inhibitors inhibited cell growth . The results also showed that both pan-HDAC and class-I-specific inhibitor treatment resulted in increased acetylation of histones , but only pan-HDAC inhibitor treatment resulted in increased tubulin acetylation , which is in agreement with their activity towards the Q9UBN7 isoform . A high-content RNAi-screening assay to identify modulators of cholesterol accumulation in Niemann-Pick type C cells . Niemann-Pick disease type C ( NPC ) is an inherited lipid storage disorder characterized by a defect in intracellular trafficking of exogenous cholesterol and glycosphingolipids . A goal for therapeutic treatment of NPC is to decrease/normalize cholesterol accumulation . We developed a functional genomics-based assay , combining high-throughput RNA interference ( HT-RNAi ) screening with high-content fluorescence imaging to identify specific genes in NPC cells that will result in more normal cholesterol levels in the diseased cells . Conditions for siRNA tranfections were optimized for 2 NPC fibroblast cell lines ( GM03123 , GM18453 ) and a normal fibroblast cell line ( GM05659 ) . RNAi screening was done using a focused-set siRNA library targeting 40 cholesterol trafficking-associated genes , knowledge mined from the existing literature on NPC disease , and/or their association with O15118 / P61916 genes . We utilized filipin staining as a measure of cholesterol accumulation in fixed NPC cells . Data analysis of these screens confirmed several genes including P01130 and P51151 that reduced cholesterol content in NPC cells . Nine genes were validated using filipin staining to detect unesterified cholesterol as well as cholesteryl BODIPY esters to study lipid trafficking . Gene silencing was also confirmed using qRT-PCR . Our results show that this technology can be applied to larger screens to identify genes responsible for lipid accumulation and/or trafficking in NPC disease , which could be instrumental in developing innovative therapies for individuals afflicted with NPC disease . Can a cocktail designed for phenotyping pharmacokinetics and metabolism enzymes in human be used efficiently in rat ? We recently designed the CIME cocktail consisting of 10 drugs to assess the activity of the major human CYPs ( P05177 , P10632 , P11712 , P33261 , P10635 and CYP3A ) , a phase II enzyme ( P22309 /6/9 ) , two drug transporters ( P-gp and Q9Y6L6 ) and a component of the renal function ( Videau et al. 2010 ) . The present work aimed at studying the usefulness of the CIME cocktail in the rat.The CIME cocktail was given per os to three male and three female rats , or incubated with rat liver microsomes . Parent substrates and metabolites were quantified by LC-MS/MS in plasma , urine and hepatic microsomal media , and phenotyping index were subsequently calculated.The CIME cocktail could therefore be used in the rat to phenotype rapidly and simultaneously CYP3A1/2 with omeprazole/omeprazole-sulfone , midazolam/1'-hydroxymidazolam or 4-hydroxymidazolam and/or dextromethorphan/3-methoxymorphinan , CYP2C6/11 with tolbutamide/4-hydroxytolbutamide , CYP2D1/2 with omeprazole/5-hydroxyomeprazole or dextromethorphan/dextrorphan , and P19224 /7 with acetaminophen/acetaminophen-glucuronide . Our results confirmed also several known gender differences and brought new information on the urinary excretion of rosuvastatin . However , the major rat CYPs , CYP2C11 and CYP2C12 , are not specifically assessed . An optimized version of the CIME cocktail should therefore be designed and would be of major importance to more largely phenotype Q09013 enzymes in rats to study Q09013 variability factors such as disease , age , or to exposure to inductors or inhibitors . Histone deacetylase inhibitors increase microRNA-146a expression and enhance negative regulation of interleukin-1β signaling in osteoarthritis fibroblast-like synoviocytes . OBJECTIVE : MiR-146a exerts negative control on inflammatory responses by suppressing cytokine-induced expression of interleukin-1 receptor-associated kinase-1 ( P51617 ) and tumor necrosis factor receptor-associated factor 6 ( Q9Y4K3 ) by impairing NF-κB activity and inhibiting the expression of target genes . Recent study suggests that histone deacetylases ( HDACs ) are involved in the regulation of microRNA ( miRNA ) expression . Therefore , we determined whether HDAC inhibitors can increase miR-146a expression , thereby inhibiting interleukin-1β ( IL-1β ) -induced signaling in osteoarthritis fibroblast-like synoviocytes ( OA-FLS ) . METHOD : MiRNA expression was analyzed using real-time PCR . IL-1β-induced downstream signals and cytokine expression were evaluated using Western blotting and ELISA . Transcription factors regulating promoter activation were identified using chromatin immunoprecipitation assays . RESULTS : IL-1β treatment of OA-FLS induced a mild ( 1.7-fold ) increase in miR-146a expression that was unable to appropriately downregulate P51617 and Q9Y4K3 expression . HDAC inhibitors , DB02546 ( vorinostat ) , and LBH589 ( DB06603 ) significantly ( 6.1- and 5.4-fold ) elevated miR-146a expression by increasing the binding of the transcription factor NF-κB to the miR-146a promoter , and negatively regulated IL-1β-induced IKK/IκB/p65 phosphorylation signaling and P05231 secretion . The increase in miR-146a expression induced by the HDAC inhibitors was prevented by transfection of miR-146a inhibitor or Q13547 ( class I HDAC ) , P56524 ( class IIa HDAC ) , and Q9UBN7 ( class IIb HDAC ) overexpression , suggesting that they were due to inhibition of HDAC activity . CONCLUSIONS : Our study demonstrated that HDAC inhibitor treatment in OA-FLS significantly increased miR-146a expression and mediated markedly negative regulation to inhibit IL-1β-induced signaling and cytokine secretion . Our results indicate the potential rationale of anti-inflammatory effects for HDAC inhibitors . DB00991 : kinetic and dynamic profile in the treatment of pain . DB00991 ( 4,5-diphenyl-2-oxazolepropionic acid ) is a non-steroidal anti-inflammatory drug ( NSAID ) which is effective in models of inflammation , pain and pyrexia . It is effective and well tolerated in the clinical management of adult rheumatoid arthritis ( RA ) , osteoarthritis ( OA ) , ankylosing spondylitis , soft tissue disorders and post operative dental pain . DB00991 has a high oral bioavailability ( 95 % ) , with peak plasma concentrations at 3 to 5 hours after dosing . It is metabolised in the liver by oxidative and conjugative pathways and readily eliminated by the renal and faecal routes . DB00991 's strong analgesic qualities are particularly useful in painful musculoskeletal conditions such as periarthritis of the shoulder , since it exhibits actions such as inhibition of P23219 and P35354 isoenzymes , inhibition of nuclear translocation of NF-kappaB and of metalloproteases , and modulates the endogenous cannabinoid system . This editorial addresses the accompanying paper by Barbara Heller and Rosanna Tarricone on the management of shoulder periarthritis pain , in which they studied the efficacy and safety of oxaprozin compared to the comparator drug diclofenac over a 15 day period . Both oxaprozin and diclofenac compared well in the primary study endpoint of reduction in shoulder pain . DB00991 and diclofenac were well tolerated and oxaprozin showed better improvement in shoulder function and in the mental health item of the SF-36 quality of life component . The study by Heller and Tarricone is an addition to the large number of clinical trials which demonstrate that oxaprozin has equal efficacy in comparison with standard doses of commonly used anti-rheumatic agents such as aspirin , diclofenac , ibuprofen , indomethacin etc. in several different painful musculoskeletal conditions . P04035 inhibitors deplete circulating classical and non-classical monocytes following human heart transplantation . BACKGROUND : Monocytes mediate immune responses following solid organ transplantation via cytokine secretion and differentiation to macrophage/dendritic cell lineages . To date , the pleiotropic immunomodulatory effect of statins on human monocytes following human heart transplantation has yet to be elucidated . This study was designed to assess the effects of statin administration on the monocyte repertoire . METHODS : 108 patients were recruited into the study . Clinical data were collected from patients ' notes . Peripheral blood immunophenotype was determined via flow cytometry ( using CD11c , P08571 , CD16 , CD49d , CD64 , P33681 and CD195 ) . RESULTS : There were fewer circulating classical ( p=0.0001 ) and non-classical ( p=0.0013 ) monocytes in patients treated with a statin . CD64 expression was down-regulated ( p=0.011 and p=0.049 ) whereas CD49d expression was up-regulated ( p=0.004 and p=0.022 ) on classical and non-classical monocytes in this group . Patients receiving DB01076 had fewer circulating classical monocytes ( p=0.001 ) compared to patients administered DB00175 . Patients receiving DB00175 had fewer circulating non-classical monocytes ( p=0.029 ) compared to patients administered DB01076 . DISCUSSION : Statin administration alters the circulating monocyte repertoire following heart transplantation , including population size , FcgammaRI and VLA-4 adhesion molecule expression . Furthermore , different statin treatments are associated with a selective depletion of macrophage or DC (re)generating monocytes . Topical glucocorticoids downregulate P23219 positive cells in nasal polyps . BACKGROUND : Influx of inflammatory cells is one of the hallmarks of nasal polyposis . As glucocorticoids ( GC ) are known to exhibit strong anti-inflammatory effects , these drugs are frequently used in the treatment of the disease . Part of the anti-inflammatory effects of GC is attributed to their interference with prostanoid synthesis . As cyclooxygenases ( P36551 ) are key enzymes in the synthesis of both pro- ( P23219 , P35354 ) and anti-inflammatory prostanoids ( P35354 ) , we investigated the role of topical GC on P23219 , P35354 and inflammatory markers in nasal polyps ( NP ) . METHODS : Immunohistochemical analysis of inflammatory markers ( P34810 , CD117 , MBP , elastase , IgE , BB-1 , P05112 , P05113 and P05231 ) , P23219 and P35354 was performed on normal nasal mucosa ( NM ) ( n = 18 ) , non-GC treated NP ( n = 27 ) and topical GC treated NP ( n = 12 ) . NP groups were matched for allergy , asthma and ASA intolerance . RESULTS : Increased numbers of eosinophils , P05113 + cells and IgE+ cells and decreased numbers of mastcells are striking features of NP inflammation ( P < 0.05 ) . In addition , increased numbers of P23219 + cells are observed in NP epithelium compared to NM ( P < 0.05 ) . CONCLUSION : Topical GC significantly reduce the number of P23219 + NP cells ( P < 0.05 ) , but have no significant effect on P35354 + NP cells . No significant reduction in the number of eosinophils is observed for GC treated NP . The number of P05113 + cells is however increased significantly upon GC treatment ( P < 0.05 ) .