interacts_with DB01268?
Stage-dependent inhibition of Plasmodium falciparum by potent Ca2+ and calmodulin modulators . The effects of Ca2+ channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W-7 and W-5 , on Plasmodium falciparum in culture were examined . Among Ca2+ blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W-7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W-5 . All Ca2+ blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 , diltiazem , calmidazolium and W-5 also retarded parasite development at earlier stages and/or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2+ and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2+ channels or P62158 . Synthesis and evaluation of ( S ) -2-(2-[18F]fluoroethoxy)-4- ( [ 3-methyl-1-(2-piperidin-1-yl-phenyl)-butyl-carbamoyl ] -methyl ) -benzoic acid ( [18F]repaglinide ) : a promising radioligand for quantification of pancreatic beta-cell mass with positron emission tomography ( PET ) . 18F-labeled non-sulfonylurea hypoglycemic agent ( S ) -2-(2-[(18)F]fluoroethoxy)-4- ( ( 3-methyl-1-(2-piperidin-1-yl-phenyl)-butylcarbamoyl ) -methyl ) -benzoic acid ( [(18)F]repaglinide ) , a derivative of the sulfonylurea-receptor ( Q09428 ) ligand repaglinide , was synthesized as a potential tracer for the non-invasive investigation of the sulfonylurea 1 receptor status of pancreatic beta-cells by positron emission tomography ( PET ) in the context of type 1 and type 2 diabetes . [(18)F] DB00912 could be obtained in an overall radiochemical yield ( RCY ) of 20 % after 135 min with a radiochemical purity higher than 98 % applying the secondary labeling precursor 2-[(18)F]fluoroethyltosylate . Specific activity was in the range of 50-60 GBq/micromol . Labeling was conducted by exchanging the ethoxy-moiety into a 2-[(18)F]fluoroethoxy group . To characterize the properties of fluorinated repaglinide , the affinity of the analogous non-radioactive (19)F-compound for binding to the human Q09428 isoform was assessed . [(19)F] DB00912 induced a complete monophasic inhibition curve with a Hill coefficient close to 1 ( 1.03 ) yielding a dissociation constant ( K(D) ) of 134 nM . Biological activity was proven via insulin secretion experiments on isolated rat islets and was comparable to that of repaglinide . Finally , biodistribution of [(18)F]repaglinide was investigated in rats by measuring the concentration of the compound in different organs after i.v. injection . Pancreatic tissue displayed a stable accumulation of approximately 0.12 % of the injected dose from 10 min to 30 min p.i . 50 % of the radioactive tracer could be displaced by additional injection of unlabeled repaglinide , indicating that [(18)F]repaglinide might be suitable for in vivo investigation with PET . New anti-angiogenic targeted therapy in advanced renal cell carcinoma ( RCC ) : current status and future prospects . OBJECTIVES : To address the rationale for anti-angiogenic targeted therapies in advanced RCC . METHODS : We reviewed the international recent literature , using Pubmed search . RESULTS : RCC is genetically linked to factors regulating angiogenesis , in particular vascular endothelial growth factor ( P15692 ) . DB01268 is a multitarget receptor tyrosine-kinase ( TK ) inhibitor , acting on P15692 receptor ( VEGFR ) and platelet-derived growth factor receptors ( P09619 ) . DB00398 is an oral multikinase inhibitor ( VEGFR and P09619 ) showing also inhibitors effect on the Raf system . Phase I trials showed no life threatening toxicities relates to these agents . Phase II and phase III trials showed that these antiangiogenic agents are effective in the treatment of advanced RCC , mainly in cytokine refractory metastatic RCC . Survival benefits exist in particular when advanced RCC patients undergo cytoreductive nephrectomies before the initiation of the systemic therapy . To better use this kind of targeted therapy in advanced RCC , different points must be developed : the identification of clinical characteristic of RCC able to predict outcomes and responses to therapy ; differences among different compounds ; advantages of combination or sequential therapies . CONCLUSIONS : Targeted therapy with DB01268 and DB00398 has been approved to FDA and is revolutioning how we clinically approach advanced RCC . DB01268 in metastatic thymic carcinomas : laboratory findings and initial clinical experience . BACKGROUND : Thymic carcinoma ( TC ) is a rare aggressive tumour . Median survival with current treatments is only 2 years . DB01268 is a multi-targeted tyrosine kinase inhibitor that has shown benefit in various other cancers . METHODS : Laboratory analyses of snap-frozen tumour tissues were performed to detect activation and genetic mutations of receptor tyrosine kinases ( RTKs ) in TC samples . On the basis of molecular analyses showing activation of multiple RTKs in their tumour , four patients with metastatic TCs refractory to conventional therapies were treated with sunitinib according to standard protocols . RESULTS : RTK analysis in three of the patients showed activation of multiple RTKs , including platelet-derived growth factor-beta and vascular endothelial growth factor 3 . Mutations of P00533 , c- P10721 , P01116 , and P15056 genes were not found . Administration of sunitinib yielded a partial remission ( lasting 2 to 18+ months ) according to the RECIST criteria in three patients and stable disease with excellent metabolic response in DB09150 -PET in another one . The overall survival with sunitinib treatment ranges from 4 to 40+ months . Withdrawal of the drug in one patient prompted rapid tumour progression that could be controlled by re-administration of sunitinib . CONCLUSIONS : DB01268 is an active treatment for metastatic TC . A panel of molecular analyses may be warranted for optimal patient selection . Survey of 548 oncogenic fusion transcripts in thyroid tumors supports the importance of the already established thyroid fusions genes . Neoplasms frequently present structural chromosomal aberrations that can alter the level of expression of a protein or to the expression of an aberrant chimeric protein . In the thyroid , the Q06710 - P37231 fusion is present in the neoplastic lesions that have a follicular architecture-follicular thyroid carcinoma ( FTC ) and follicular variant of papillary thyroid carcinoma ( FVPTC ) , and less frequently in follicular thyroid adenoma ( DB00499 ) , while the presence of P07949 /PTC fusions are largely restricted to papillary thyroid carcinoma ( PTC ) . The ability to detect fusion genes is relevant for a correct diagnosis and for therapy . We have developed a new fusion gene microarray-based approach for simultaneous analysis of all known and predicted fusion gene variants . We did a comprehensive screen for 548 known and putative fusion genes in 27 samples of thyroid tumors and three positive controls-one thyroid cancer cell line ( TPC-1 ) and two PTCs with known Q16204 - P07949 ( alias P07949 / Q13635 ) fusion gene , using this microarray . Within the thyroid tumors tested , only well known , previously reported fusion genes in thyroid oncology were identified . Our results reinforce the pathogenic role played by P07949 / Q13635 , P07949 /PTC3 , and Q06710 - P37231 fusion genes in thyroid tumorigenesis . Disease stabilization of progressive olfactory neuroblastoma ( esthesioneuroblastoma ) under treatment with sunitinib mesylate . Olfactory neuroblastoma ( esthesioneuroblastoma ) is a rare neoplasm of the olfactory epithelium in the upper nasal cavity . Here , we report the case of a 69-year-old man who presented with massive progression of a metastatic esthesioneuroblastoma after endonasal resection , functional neck dissection , and radiotherapy of local and distant tumor relapses . After exhaustion of all conventional therapeutic options , we initiated treatment with the oral multityrosinekinase inhibitor sunitinib mesylate . Using this drug , significant improvement of clinical symptoms , disease stabilization , and recovery from Karnofsky index of 40 % to 70 % could be achieved in the absence of significant adverse drug effects . The patient died 15 months after initiation of sunitinib therapy due to complications of a traumatic femoral neck fracture without evidence of tumor progression . Immunohistochemical analysis of tumor tissue specimens obtained at initial surgery revealed ample expression of platelet-derived growth factor receptor ( P09619 ) -b on stromal and endothelial cells . DB01268 should be considered for palliative therapy of advanced esthesioneuroblastoma . DB03203 kinase-1 activation causes acquired resistance against DB01268 in renal cell carcinoma cells . Multi-target tyrosine kinase inhibitor DB01268 has been widely used in cancer treatment , including metastatic renal cell carcinoma . However , most patients who initially benefit from DB01268 develop resistance with extended usage of DB01268 , which is referred to as " acquired resistance " . The molecular mechanisms contributing to this acquired resistance remain poorly understood . In this present study , we established DB01268 -resistant cell lines from human renal cell lines ( 786-O , A498 , ACHN and CAKI1 ) by continuous treatment with DB01268 to explore the molecular mechanism leading to DB01268 resistance . We found that P09619 -β expression in cell seems to be a protective factor against DB01268 resistance formation . In addition , we found that both SK1 and P29323 were activated in DB01268 -resistance cell lines and SK1 and P29323 inhibitors could resensitize DB01268 -resistant cell lines . In conclusion , our observations suggest that SK1 and P29323 activation is a feature of resistant cell lines , which serves as an alternative pathway evading anti-tumor activity of DB01268 . Genome-wide association study identifies genetic determinants of warfarin responsiveness for Japanese . DB00682 is a commonly used anticoagulant , whose dose needs to be determined for each individual patient owing to large inter-individual variability in its therapeutic dose . Although several clinical and genetic variables influencing warfarin dose have been identified , uncovering additional factors are critically important for safer use of warfarin . Through a genome-wide association study , we identified single-nucleotide polymorphism ( SNP ) rs2108622 [ cytochrome P450 , family 4 , subfamily F , polypeptide 2 ( P78329 ) ] as a genetic determinant of warfarin responsiveness for Japanese . Stratifying subjects who have been pre-classified according to the genotypes of SNP rs10509680 [ cytochrome P450 , family 2 , subfamily C , polypeptide 9 ( P11712 ) ] and SNP rs9923231 [ vitamin K epoxide reductase complex subunit 1 ( Q9BQB6 ) ] , based on their genotypes of rs2108622 allowed identification of subjects who require higher dose of warfarin . Incorporating genotypes of rs2108622 into a warfarin dosing algorithm that considers age , body surface area , status of amiodarone co-administration and genotypes of SNPs in the P11712 and Q9BQB6 genes improved the model 's predictability to 43.4 % . In this study , the association of P78329 with warfarin dose of the Japanese has been established for the first time . Besides , a warfarin dosing algorithm that incorporates genotypes of rs2108622 and amiodarone co-administration status was suggested for the Japanese . Our study also implied that common SNPs other than those in the P11712 , Q9BQB6 and P78329 genes that show strong effect on the therapeutic warfarin dose might not exist . DB01268 as a second-line therapy for advanced GISTs after failure of imatinib : relationship between efficacy and tumor genotype in Korean patients . BACKGROUND : To assess the efficacy and safety of sunitinib with regards to primary genotypes of tumor in Korean patients with advanced gastrointestinal stromal tumors ( GISTs ) who failed an initial therapy of imatinib . METHODS : Clinical data were collected from 88 consecutive patients with metastatic/unresectable GISTs treated with sunitinib at the Asan Medical Center . RESULTS : The median time-to-progression ( TTP ) and overall survival ( OS ) times were 7.1 months and 17.6 months , respectively . Of the 74 patients tested for P10721 ( exons 9 , 11 , 13 , 17 ) and P16234 ( exons 12 and 18 ) , patients with P10721 exon 9 mutant GIST ( n = 11 , 14.9 % ) showed numerically better clinical benefit ( objective response or stable disease ≥ 24 weeks ) rate ( 63.6 % vs 46.8 % , p = 0.504 ) and TTP ( median 13.6 mo vs 6.9 mo , p = 0.631 ) than those with P10721 exon 11 mutant GIST ( n = 47 , 63.5 % ) . The most common grade 3/4 adverse events included neutropenia ( 34.1 % ) , thrombocytopenia ( 33.0 % ) and hand-foot skin reaction ( 25.0 % ) . CONCLUSIONS : DB01268 is an effective and safe second-line therapy for Korean patients with advanced GIST . The superior efficacy of sunitinib against GISTs with P10721 exon 9 mutations appears to be similar in Korean patients to Western experience although statistical significance was not secured . [ GIST refractory to imatinib treatment ] . After the introduction of imatinib , the outcome for patients with advanced gastro-intestinal stromal tumor ( GIST ) was vastly improved . However , resistance to imatinib has become a new problem . The cause of resistance to imatinib is the low sensitivity of gene mutations in the P10721 gene or PDGFRα , or an acquisition of additional mutations , and a low plasma level of imatinib . DB01268 is the first drug that showed an effectiveness for treating GIST refractory to imatinib . New molecular target drugs for overcoming imatinib resistance are being developed now . DB01268 acts primarily on tumor endothelium rather than tumor cells to inhibit the growth of renal cell carcinoma . DB01268 is a broad-spectrum small-molecule inhibitor of receptor tyrosine kinases ( RTK ) that serves as the present standard of care for first-line therapy of advanced clear cell renal cell carcinoma ( ccRCC ) . A full understanding of the targets and mechanism of action of sunitinib in ccRCC treatment remains incomplete . In this study , we evaluated several tumor cell and endothelial targets of sunitinib and investigated which RTK(s) may specifically contribute to its therapeutic effects . Microarray expression profiling and Western blot analysis revealed that among known sunitinib targets , only platelet-derived growth factor receptor-beta and vascular endothelial growth factor receptor-2 ( P35968 ) were overexpressed in ccRCCs relative to normal tissues . DB01268 was unable to inhibit survival or proliferation of ccRCC cells at pharmacologically relevant concentrations ( approximately 0.1 micromol/L ) that inhibit RTK targets . In contrast , sunitinib inhibited endothelial cell proliferation and motility at the same concentrations by suppressing P35968 signaling . Moreover , whereas sunitinib inhibited the growth of ccRCC xenograft tumors and decreased tumor microvessel density as soon as 12 hours after treatment , sunitinib showed no significant effects on tumor cell proliferation or apoptosis up to 72 hours after treatment . Our findings indicate that sunitinib inhibits ccRCC growth primarily through an antiangiogenic mechanism and not through direct targeting of ccRCC tumor cells . Alternative dosing schedules for sunitinib as a treatment of patients with metastatic renal cell carcinoma . DB01268 malate ( Sutent ; Pfizer , Inc. , New York , NY ) is an oral multitargeted tyrosine kinase inhibitor that inhibits P15692 receptors ( P17948 , P35968 , and P35916 ) among a family of kinase targets and have a central role in first-line treatment of metastatic renal cell carcinoma ( mRCC ) . The approved schedule for sunitinib is 50mg/day oid in the so called " 4 weeks on and two weeks off " ( 4/2 schedule ) . Since treatment with sunitinib can be maintained for years , adequate treatment of adverse events ( AEs ) and care for quality of life is essential . For this reason , several alternative schedules have been proposed in order to personalize sunitinib administration and reduce related toxicity . This review discusses the efficacy and tolerability of alternative regimens to the standard 4/2 schedule that have been investigated in RCC patients including schedule of 50mg/day 2-weeks on/1-week off , continuous schedule of 37.5mg daily and the " Stop and Go strategy " . Crystal structure of the PDZ1 domain of human Na(+)/H(+) exchanger regulatory factor provides insights into the mechanism of carboxyl-terminal leucine recognition by class I PDZ domains . The Na(+)/H(+) exchanger regulatory factor ( O14745 ; also known as O14745 ) contains two PDZ domains that mediate the assembly of transmembrane and cytosolic proteins into functional signal transduction complexes . The O14745 PDZ1 domain interacts specifically with the motifs DSLL , DSFL , and DTRL present at the carboxyl termini of the beta(2) adrenergic receptor ( beta(2)AR ) , the platelet-derived growth factor receptor ( P09619 ) , and the cystic fibrosis transmembrane conductance regulator ( P13569 ) , respectively , and plays a central role in the physiological regulation of these proteins . The crystal structure of the human O14745 PDZ1 has been determined at 1.5 A resolution using multiwavelength anomalous diffraction phasing . The overall structure is similar to known PDZ structures , with notable differences in the O14745 PDZ1 carboxylate-binding loop that contains the GYGF motif , and the variable loop between the beta2 and beta3 strands . In the crystalline state , the carboxyl-terminal sequence DEQL of PDZ1 occupies the peptide-binding pocket of a neighboring PDZ1 molecule related by 2-fold crystallographic symmetry . This structure reveals the molecular mechanism of carboxyl-terminal leucine recognition by class I PDZ domains , and provides insights into the specificity of O14745 interaction with the carboxyl termini of several membrane receptors and ion channels , including the beta(2)AR , P09619 , and P13569 . Emergence of polyclonal P36888 tyrosine kinase domain mutations during sequential therapy with sorafenib and sunitinib in P36888 -ITD-positive acute myeloid leukemia . PURPOSE : To evaluate the clinical activity of sequential therapy with sorafenib and sunitinib in P07333 -like tyrosine kinase 3 ( P36888 ) -internal tandem duplication ( ITD ) -positive acute myelogenous leukemia ( AML ) and monitor the emergence of secondary P36888 tyrosine kinase domain ( TKD ) mutations during treatment . EXPERIMENTAL DESIGN : Six children with relapsed/refractory AML were treated with sorafenib in combination with clofarabine and cytarabine , followed by single-agent sorafenib if not a candidate for transplantation . DB01268 was initiated after sorafenib relapse . Bone marrow samples were obtained for assessment of P36888 TKD mutations by deep amplicon sequencing . The phase of secondary mutations with ITD alleles was assessed by cloning and sequencing of P36888 exons 14 through 20 . Identified mutations were modeled in Ba/ P13726 cells , and the effect of kinase inhibitors on P36888 signaling and cell viability was assessed . RESULTS : Four patients achieved complete remission , but 3 receiving maintenance therapy with sorafenib relapsed after 14 to 37 weeks . DB01268 reduced circulating blasts in two patients and marrow blasts in one . Two patients did not respond to sorafenib combination therapy or sunitinib . P36888 mutations at residues D835 and F691 were observed in sorafenib resistance samples on both ITD-positive and -negative alleles . Deep sequencing revealed low-level mutations and their evolution during sorafenib treatment . DB01268 suppressed leukemic clones with D835H and F691L mutations , but not D835Y . Cells expressing sorafenib-resistant P36888 mutations were sensitive to sunitinib in vitro . CONCLUSIONS : DB01268 has activity in patients that are resistant to sorafenib and harbor secondary P36888 TKD mutations . The use of sensitive methods to monitor P36888 mutations during therapy may allow individualized treatment with the currently available kinase inhibitors . Cytokeratin expression in gastrointestinal stromal tumors : morphology , meaning , and mimicry . BACKGROUND : Gastrointestinal stromal tumors ( GISTs ) are biologically distinctive neoplasms harboring P10721 and P16234 mutations . Cytokeratin expression in GISTs is an under-recognized diagnostic pitfall , especially in high grade GISTs with limited biopsy material and from metastatic sites . MATERIALS AND METHODS : We evaluated the histomorphology and expression of four ' broad-spectrum ' cytokeratin markers , P02730 - P48751 , P62158 5.2 , P85037 -116 , and 34β Q96BH3 in 64 GISTs diagnosed over a 68-month period . Individual cytokeratins 5 , 6 , 7 , 8 , 14 , 17 , 18 , 19 , and 20 were investigated in the ' broad-spectrum ' cytokeratin-positive GISTs . RESULTS : Of 64 GISTs , 10 ( 15 % ) demonstrated cytokeratin immunopositivity . All 10 , considered high risk by the National Institutes of Health consensus approach , were immunopositive for P62158 5.2 and P85037 -116 . Seven were P02730 - P48751 immunopositive . Cytokeratins 8 and 18 were confirmed in 10 and 9 GISTs , respectively . One GIST demonstrated biphasic morphology with cytokeratin immunonegativity in low-grade spindle and immunopositivity in high-grade epithelioid foci . P10721 and P16234 mutational analysis , undertaken in 5/10 cytokeratin-positive GISTs , harbored P10721 exon 11 mutations . CONCLUSION : We hypothesize that cytokeratin expression exclusively in high risk GISTs is a consequence of tumor progression . Given the increasing number of commercially available broad-spectrum cytokeratin immunomarkers , including those reacting with cytokeratins 8 and 18 , cytokeratin-positive GISTs must be differentiated from carcinomas , melanomas , and a range of cytokeratin-positive sarcomas to ensure optimal patient management and prognostication . DB01268 and periodic hair depigmentation due to temporary c- P10721 inhibition . Comparison of two polymer-based immunohistochemical detection systems : ENVISION+ and ImmPRESS . The non-specific background reaction produced in avidin-biotin-based immunohistochemistry , particularly after harsh antigen retrieval procedures , has promoted the use of non-avidin-biotin systems , yet there are few reports comparing the performance of non-avidin-biotin , polymer-based methods . In this study we compare two of these methods , ENVISION+trade mark and ImmPRESS , in animal tissues . We examined the immunoreactivity of 18 antigens in formalin-fixed , paraffin-embedded tissues . Antigens were located in the cytoplasmic membrane ( CD11d , P05107 and CD79a ) , cytoplasm ( calretinin , P23219 , P35354 , Glut-1 , HepPar 1 , P10721 , Melan A , tryptase and uroplakin III ) or nucleus ( Q2TAK8 , P09936 and thyroid transcription factor 1 ) . We also evaluated three infectious agents ( Aspergillus , calicivirus and West Nile virus ) . The staining with ENVISION+ or ImmPRESS was performed simultaneously for each antigen . The intensity of the reaction and background staining were scored . ImmPRESS yielded similar or higher reaction intensity than ENVISION+trade mark in 16/18 antigens . ImmPRESS produced abundant background with the other two antigens ( calretinin and P35354 ) , which hindered interpretation of the specific reaction . The cost of ImmPRESS was 25 % lower than for ENVISION+trade mark . Based on these results , ImmPRESS is a good polymer-based detection system for routine immunohistochemistry . Expression and mutational status of treatment-relevant targets and key oncogenes in 123 malignant salivary gland tumours . BACKGROUND : Malignant tumours of the salivary glands ( MSGT ) are rare and pleomorphic entities . Patients with advanced disease may benefit from targeted therapy ; however , specific targets for optimising and personalising treatments are yet to be identified . DESIGN : Immunohistochemistry for C- P10721 , P00533 , P04626 , P15941 , phospho- P42345 , androgen/estrogens/progesterone receptors and Ki67 was carried out and evaluated in terms of progression-free and overall survival . High throughput molecular screening of key oncogenes was done in 107 patients using routine diagnostic methods and Sequenom technology . RESULTS : Several therapy leads were identified , including high levels of P04626 and androgen receptors in salivary duct carcinomas , C- P10721 in myoepithelial carcinomas and P00533 in mucoepidermoid carcinomas . Recurrent mutations involving downstream elements of the P00533 pathway were found in P01112 , notably in tumours with a myoepithelial component , and in other key oncogenes ( P01116 / P01111 /PI3KCA/ P15056 /MAP2K ) . On the other hand , < 1 % of samples had P00533 or P04626 mutations . CONCLUSION : Several tumour subtypes overexpressed targets of directed therapies suggesting potential therapy leads . Genotyping results suggest activation downstream of P00533 in 18 of the 107 samples that could be associated with low efficacy of P00533 inhibitors . Other molecules , such as PI3K/MEK or P42345 inhibitors , may have anti-tumour activity in this subgroup . The high mutation rate in P01112 highlights a novel key oncogenic event in MSGT . First report of warfarin dose requirements in patients possessing the P11712 *12 allele . BACKGROUND : DB00682 is the most frequently prescribed anticoagulant in North America and Europe . It is administered as a racemate , but S-warfarin is principally responsible for its anticoagulant activity . Cytochrome P450 ( CYP ) 2C9 is the enzyme primarily responsible for the metabolism of S-warfarin . Numerous variant alleles of P11712 have been identified . The P11712 *12 ( rs9332239 ) allele harbors a P489S substitution in P11712 which has been shown to result in a 40 % decline in catalytic activity in vitro . CASES : Four Caucasian patients with a low mean weekly warfarin dose ( MWWD ) were genotyped for P11712 , Q9BQB6 and P02649 variant alleles . None of the four patients carried the common P11712 variant alleles ( *2 , *3 , *5 , *6 , *7 , *8 , *9 , *11 , *13 ) despite a relatively low MWWD ( 23.4±7.94 mg ) compared to 208 patients carrying the CYP29C9*1 genotype ( 32.2±12.65 mg ) . Given that P11712 *12 confers decreased in vitro activity to the enzyme , we investigated whether these patients carried this allele . All four patients were P11712 *12 CT heterozygotes . Individual comparisons with patients possessing the same Q9BQB6 and P02649 genotypes also demonstrated lower dose requirements in the patients that possessed P11712 *12 allele . CONCLUSIONS : There are no reports of the clinical impact of rs9332239 on P11712 substrates . This is the first report of patients with the rare P11712 *12 genotype and lower warfarin dose requirements . Expression and release of phosphatidylinositol anchored cell surface molecules by a cell line derived from sensory neurons . Early postnatal mouse dorsal root ganglion neurons were found to express several glycosylphosphatidylinositol-anchored ( P06744 ) molecules from the immunoglobulin superfamily ( neural cell adhesion molecule 120 kD isoform , P13726 , Thy1 ) whose expression is developmentally regulated . A hybrid cell line ( ND26 ) , made by fusing postmitotic rat dorsal root ganglion ( Q86YR7 ) neurons with the mouse neuroblastoma N18Tg2 , could be induced to differentiate by manipulating the composition of the culture medium and expressed similar P06744 molecules to Q86YR7 neurons . We used this model system to investigate the metabolism of P06744 -anchored molecules . We found that neural cell adhesion molecule 120 Kd isoform expression decreased upon differentiation , whereas the level of P13726 and Thy1 increased , suggesting a role in neurite outgrowth processes . The ratio of molecules cleavable by exogenous phosphatidylinositol phospholipase C ( PI- P98160 ) was similar for all the P06744 -anchored molecules , which could mean that cell-specific modifications of the basic anchoring structure determine the level of potentially releasable molecules . Measurements of spontaneous release indicated that this reflected the overall level of expression of these molecules by the ND26 cell line . Finally , we observed an effect of dibutyryl DB02527 on the level of expression of P13726 and Thy1 but not of N- P62158 . However , we could not detect any significant effect of nerve growth factor ( P01138 ) either on the level of expression or on the amount of spontaneously released molecules . P01308 secretory defects and impaired islet architecture in pancreatic beta-cell-specific P40763 knockout mice . Normal islet formation and function depends on the action of various growth factors operating in pre- and postnatal development ; however , the specific physiological function of each factor is largely unknown . Loss-of-function analyses in mice have provided little information so far , perhaps due to functional redundancies of the growth factors acting on the pancreas . The present study focuses on the role of the transcription factor P40763 in insulin-producing cells . P40763 is one of the potential downstream mediators for multiple growth factors acting on the pancreatic beta-cells , including betacellulin , hepatocyte growth factor , growth hormone , and heparin-binding P01133 -like growth factor . To elucidate its role in the beta-cells , the P40763 gene was disrupted in insulin-producing cells in mice ( P40763 -insKO ) , using a cre-mediated gene recombination approach . Unexpectedly , P40763 -insKO mice exhibited an increase in appetite and obesity at 8 weeks of age or older . The mice showed partial leptin resistance , suggesting that expression of the RIP ( rat insulin promoter ) -cre transgene in hypothalamus partially inhibited the appetite-regulating system . Intraperitoneal glucose tolerance tests , performed in non-obese 5-week-old mice , showed that the P40763 -insKO mice were glucose intolerant . Islet perifusion experiments further revealed a deficiency in early-phase insulin secretion . Whereas islet insulin content or islet mass was not affected , expression levels of P11168 , Q09428 , and P15692 were significantly reduced in P40763 -insKO islets . Interestingly , P40763 -insKO mice displayed impaired islet morphology : alpha-cells were frequently seen in central regions of islets . Our present observations demonstrate a unique role of P40763 in maintaining glucose-mediated early-phase insulin secretion and normal islet morphology . New aspects in the pathogenesis of diabetic atherothrombosis . Diabetes mellitus is increasing worldwide , resulting from the interaction of obesity , inflammation , and hyperglycemia . Activated immunity and cytokine production lead to insulin resistance and other components of the metabolic syndrome , establishing the link between diabetes and atherosclerosis . Hyperglycemia-induced endothelial dysfunction is mediated by increased oxidative stress , a promoter of adventitial inflammation and vasa vasorum neovascularization in experimental models of diabetic atherosclerosis . Recent studies have documented increased inflammation , neovascularization , and intraplaque hemorrhage in human diabetic atherosclerosis . This inflammatory microangiopathic process is independently associated with plaque rupture , leading to coronary thrombosis . P13726 , the most potent trigger of the coagulation cascade , is increased in diabetic patients with poor glycemic control . Circulating tissue factor microparticles are also associated with apoptosis of plaque macrophages , closing the link among inflammation , plaque rupture , and blood thrombogenicity . High-density lipoproteins , responsible for free cholesterol removal , are reduced in patients with insulin resistance and diabetes . High-density lipoprotein therapy leads to a significant decrease in plaque macrophages and increase in smooth-muscle cells . These beneficial effects may be responsible for coronary plaque stabilization in patients treated with recombinant P02647 Milano/phospholipid complex . Finally , peroxisomal proliferator-activated receptors ( PPARs ) are now considered the nuclear transcriptional regulators of atherosclerosis . Three subfamilies , including Q07869 , -delta , and -gamma , have been identified with crucial roles in lipid metabolism , plaque inflammation , expression of adhesion molecules and cytokines , and regulation of matrix metalloproteinases . Multiple experimental studies have documented plaque stabilization with P37231 agonists , a group of medications holding great promise in the treatment of diabetes atherosclerosis . AL3810 , a multi-tyrosine kinase inhibitor , exhibits potent anti-angiogenic and anti-tumour activity via targeting VEGFR , FGFR and P09619 . Angiogenesis plays an important role in neoplastic transformation and progression as well as in the metastasis process of most human cancers . Herein , we identified AL3810 as a novel and orally bioavailable small molecular inhibitor with potent inhibitory activity against multiple tyrosine kinases involved in the process of angiogenesis . We found that AL3810 substantially inhibited the autophosphorylation of P35968 , PDGFRβ and P11362 in endothelial cells . Moreover , AL3810 exhibited potent anti-angiogenesis activity , manifested by significant inhibition of microvessel outgrowth of rat arterial ring and chickallantochorion membrane ( P62158 ) in ex vivo angiogenesis models . Daily dosing of AL3810 has shown broad-spectrum anti-tumour activity in human kidney , pancreas , liver cancer xenograft models . Importantly , immunohistochemistry results demonstrated that the anti-tumour activity of AL3810 was closely correlated with its anti-angiogenesis activity , as demonstrated by a decreased microvessel area and reduced microvessel numbers in tumour tissues . The overall pharmacological profiles of AL3810 are superior to sorafenib . The clinical trials of AL3810 will soon be launched in China . P37231 ligands are potent inhibitors of angiogenesis in vitro and in vivo . P37231 ( PPARgamma ) is a nuclear receptor that functions as a transcription factor to mediate ligand-dependent transcriptional regulation . Activation of PPARgamma by the naturally occurring ligand , 15-deoxy-Delta12,14-prostaglandin J2 ( 15d-PGJ2 ) , or members of a new class of oral antidiabetic agents , e.g. BRL49653 and ciglitizone , has been linked to adipocyte differentiation , regulation of glucose homeostasis , inhibition of macrophage and monocyte activation , and inhibition of tumor cell proliferation . Here we report that human umbilical vein endothelial cells ( HUVEC ) express PPARgamma mRNA and protein . Activation of PPARgamma by the specific ligands 15d-PGJ2 , BRL49653 , or ciglitizone , dose dependently suppresses HUVEC differentiation into tube-like structures in three-dimensional collagen gels . In contrast , specific PPARalpha and -beta ligands do not affect tube formation although mRNA for these receptors are expressed in HUVEC . PPARgamma ligands also inhibit the proliferative response of HUVEC to exogenous growth factors . Treatment of HUVEC with 15d-PGJ2 also reduced mRNA levels of vascular endothelial cell growth factor receptors 1 ( Flt-1 ) and 2 ( Flk/ P35968 ) and urokinase plasminogen activator and increased plasminogen activator inhibitor-1 ( P05121 ) mRNA . Finally , administration of 15d-PGJ2 inhibited vascular endothelial cell growth factor-induced angiogenesis in the rat cornea . These observations demonstrate that PPARgamma ligands are potent inhibitors of angiogenesis in vitro and in vivo , and suggest that PPARgamma may be an important molecular target for the development of small-molecule inhibitors of angiogenesis . The effects of five alkaloids from Bulbus Fritillariae on the concentration of DB02527 in P29320 cells transfected with muscarinic M(2) receptor plasmid . The aim of this study was to investigate the effects of five alkaloids , namely verticine , verticinone , imperialine , imperialine-3beta-D-glucoside , and puqietinone , purified from Bulbus Fritillariae and used as an antitussive drug in traditional Chinese medicine , on their antimuscarinic M(2) function and the DB02527 level of P29320 cells transfected with muscarinic M(2) receptor plasmid . By transfecting the P29320 cells with the method of calcium phosphate co-precipitation and screening with G418 , the cells stably expressing M(2) receptor were identified . The expression of M(2) receptor in P29320 cells was confirmed by both RT-PCR and western blot . The DB02527 level in the treated cells was analyzed with RIA method ( (125)I- DB02527 P10721 ) . And the results suggested that the five alkaloids could significantly elevate the DB02527 concentration in the P29320 cells transfected with muscarinic M(2) receptor plasmid ( p < 0.01 ) . Endothelial Q09428 -8 acts in an P29323 -independent pathway during atrioventricular cushion development . Q09428 -8 , a conserved leucine-rich repeats protein , was first identified as a positive regulator of Ras pathway in Caenorhabditis elegans . Biochemical studies indicated that Q09428 -8 interacts with Ras and Raf , leading to the elevated P29323 activity . However , the physiological role of Q09428 -8 during mammalian development remains unclear . Here we found that germline deletion of Q09428 -8 in mice resulted in early embryonic lethality . Inactivated Q09428 -8 specifically in mouse endothelial cells ( ECs ) revealed that Q09428 -8 is essential for embryonic heart development . Q09428 -8 deficiency in ECs resulted in late embryonic lethality , and the mutant mice displayed multiple cardiac defects . The reduced endothelial-mesenchymal transformation ( EMT ) and the reduced mesenchyme proliferation phase were observed in the atrioventricular canal ( AVC ) within the mutant hearts , leading to the formation of hypoplastic endocardial cushions . However , P29323 activation did not appear to be affected in mutant ECs , suggesting that Q09428 -8 may act in an P29323 -independent pathway to regulate AVC development . Heterogeneous expression of cyclooxygenase-2 and inducible nitric oxide synthase within colorectal tumors : correlation with tumor angiogenesis . BACKGROUND : Recent studies have shown that the cyclooxygenase ( P36551 ) and the inducible nitric oxide synthase ( P35228 ) pathways are involved in the development of tumor angiogenesis in human cancers . AIMS : To investigate whether a different pattern of P35354 and P35228 expression/activity exists within different areas of colorectal tumors and to analyze the relationship between these two enzymes and tumor angiogenesis . METHODS : Microvessel density ( P53602 ) and P35354 , P35228 , vascular endothelial growth factor ( P15692 ) and P15692 receptor-2 ( P35968 ) protein expression were evaluated at both the invasive front ( IF ) and the tumor center ( TC ) in 46 human colorectal cancer specimens . We also investigated the concentration of DB00917 and NO at the same sites . RESULTS : P35354 and P35228 protein expression and activity were significantly higher within the IF than the TC of the tumor specimens . Similarly , P53602 and P15692 / P35968 expression significantly increased from the TC to the IF . Only P35354 expression was significantly correlated with P53602 and P15692 / P35968 expression at both the TC and the IF . CONCLUSION : Our study shows a heterogeneous expression of P35354 and P35228 in colorectal cancer . The up-regulation of P35354 at the IF parallels an increase in vessel density and P15692 / P35968 expression , thus supporting the hypothesis that the tumor periphery is the most aggressive portion of a colorectal tumor . DB00316 -inhibitable P35354 . Although paracetamol potently reduces pain and fever , its mechanism of action has so far not been satisfactorily explained . It inhibits both P23219 and P35354 weakly in vitro , but reduces prostaglandin synthesis markedly in vivo . In mouse macrophage J774.2 cells , P35354 induced for 48 hr with high concentrations of NSAIDs is more sensitive to inhibition with paracetamol than endotoxin-induced P35354 . In the rat pleurisy model of inflammation , a second peak of P35354 protein appears 48 hr after administration of the inflammatory stimulus , during the resolution phase of the inflammatory process . Inhibition of the activity of this late-appearing P35354 with indomethacin or a selective P35354 inhibitor , delays resolution and the inflammation is prolonged . Cultured lung fibroblasts also express P35354 activity after stimulation with IL-1beta which is highly sensitive to inhibition with paracetamol . Thus , evidence is accumulating for the existence of a P35354 variant or a new P36551 enzyme which can be inhibited with paracetamol . Pharmacological management of gastrointestinal stromal tumours : an update on the role of sunitinib . The efficacy and tolerability of the receptor tyrosine kinase inhibitor , sunitinib malate , have been demonstrated in phase I-III clinical trials of patients with imatinib-resistant or imatinib-intolerant gastrointestinal stromal tumours ( GIST ) as well as in a worldwide expanded-access study and in a continuous daily dosing ( CDD ) trial . Tumour genotype may have a significant influence on the activity of sunitinib in patients with imatinib-resistant GIST . DB01268 activity was observed across different GIST genotypes and particularly in patients with wild-type and P10721 exon 9 mutations ( all relatively resistant to standard-dose imatinib ) and in patients with secondary P10721 exons 13 and 14 mutations . Adverse events with sunitinib were generally mild to moderate and easily managed by dose reduction , dose interruption or standard supportive measures . Treatment discontinuation can be avoided in most patients by close monitoring before and during treatment with appropriate adverse event management as necessary . The correlation between treatment exposure and clinical response is prompting the search for new approaches to treatment optimisation to ensure that patients derive maximum benefit from sunitinib therapy , including dose adjustments based on blood testing to ensure optimal drug exposure , and the use of the alternative CDD regimen to avoid treatment interruption . Inhibition of activated receptor tyrosine kinases by DB01268 induces growth arrest and sensitizes melanoma cells to DB00188 by blocking Akt pathway . Despite the use of multiple therapeutic strategies , metastatic melanoma remains a challenge for oncologists . Thus , new approaches using combinational treatment may be used to try to improve the prognosis of this disease . In this report , we have analyzed the expression of receptor tyrosine kinases ( RTKs ) in melanoma specimens and in four metastatic melanoma cell lines . Both melanoma specimens and cell lines expressed RTKs , suggesting that they may represent eventual targets for multitargeted tyrosine kinase inhibitor , Suntinib . DB01268 reduced the proliferation of two melanoma cell lines ( M16 and M17 ) and increased apoptosis in one of them ( M16 ) . Moreover , the two metastatic melanoma cell lines harbored an activated receptor ( PDGFRα and VEGFR , respectively ) , and DB01268 suppressed the phosphorylation of the RTKs and their downstream targets Akt and ribosomal protein S6 , in these two cell lines . Similar results were obtained when either PDGFRα or P35968 expression was silenced by lentiviral-mediated short-hairpin RNA delivery in M16 and M17 , respectively . To evaluate the interaction between DB01268 and DB00188 , median dose effect analysis using MTT assay was performed , and combination index was calculated . DB00188 synergistically enhanced the DB01268 -induced growth arrest in DB01268 -sensitive cells ( combination index < 1 ) . Moreover , LY294002 , a PI3K inhibitor , sensitized melanoma cells to DB00188 treatment , suggesting that downregulation of phospho-Akt by DB01268 mediates the synergy obtained by DB00188 + DB01268 cotreatment . Altogether , our results suggest that melanoma cells harboring an activated RTK may be clinically responsive to pharmacologic RTK inhibition by DB01268 , and a strategy combining DB01268 and DB00188 , may provide therapeutic benefit . Growth factors expression in patients with erosive esophagitis . Although the pathogenesis and treatment of erosive esophagitis ( EE ) is well recognized , little is known about the cellular and molecular mechanisms of mucosal healing in EE patients . In this pilot study , we enrolled typical EE patients to evaluate what kinds of growth factors and their receptors were activated in their injured esophageal mucosa . Forty endoscopically proved EE patients were consecutively enrolled . Messenger RNA expressions , which includes keratinocyte growth factor ( KGF ) and its receptor ( P21802 ) , epidermal growth factor ( P01133 ) and its receptor ( P00533 ) , hepatocyte growth factor ( P14210 ) and its receptor ( HGFR ) , basic fibroblast growth factor ( P09038 ) , vascular endothelial growth factor ( P15692 ) , and cyclooxygenase ( P36551 ) -1 and P35354 , were measured using real-time polymerase chain reaction ( PCR ) . Data were compared between the injured EE mucosa and their normal esophageal mucosa above EE . The mRNA expressions of P14210 , HGFR , P01133 , P15692 , and P35354 , but not P00533 , KGF , P21802 , P09038 , and P23219 , were significantly increased in the injured mucosa of EE patients compared with those of normal mucosa ( P < 0.05 ) . The study found that P14210 , HGFR , P01133 , P15692 , and , P35354 are activated in the injured mucosa of EE patients ; their activation might be involved in mucosal repair and ulcer healing of EE . Preclinical evaluation of sunitinib as single agent or in combination with chemotherapy in nasopharyngeal carcinoma . PURPOSE : DB01268 is a multi-target receptor tyrosine kinase ( RTK ) inhibitor against vascular endothelial growth factor receptors , platelet-derived growth factor receptors ( P09619 ) , c-kit and P07949 . Several of these RTKs are known to be involved in the progression of nasopharyngeal carcinoma ( NPC ) . Here , we evaluated the preclinical activities of sunitinib in NPC . METHOD : We determined the basal level of total and phosphorylated P09619 , c-kit and P07949 by immunoblotting in a panel of five NPC cell lines . The effect of sunitinib on NPC cell proliferation was evaluated by MTT assay . We further studied the effect of sunitinib on NPC cell cycle progression and apoptosis . We investigated the in vitro and in vivo activities of sunitinib as single agent and in combination with cisplatin or docetaxel in NPC cell lines and tumor xenografts . RESULTS : DB01268 exhibited dose-dependent growth inhibition in all NPC cell lines tested with IC(50) between 2-7.5 μM and maximum inhibition of over 97 % . DB01268 induced apoptosis and cell cycle arrest at G(0)/G(1) phase . In vitro , sunitinib moderately enhanced the growth inhibition of cisplatin or docetaxel . Single agent sunitinib demonstrated significant growth inhibition , reduced microvessel density and caused extensive tumor necrosis in a NPC xenograft model . However , concurrent administration of sunitinib and docetaxel induced severe toxicity in mice without enhanced antitumor effect . CONCLUSIONS : Single agent sunitinib demonstrated potent in vitro and in vivo growth inhibition in NPC . When combined with chemotherapy , sequential instead of concurrent administration schedule should be further explored . Pharmacologic modulation of niche accessibility via tyrosine kinase inhibition enhances marrow and thymic engraftment after hematopoietic stem cell transplantation . Essential survival signals within hematopoietic stem cell ( P19526 ) and thymic niches are mediated by receptor tyrosine kinases , which can be reversibly inhibited using clinically available drugs . We studied whether sunitinib , a multityrosine kinase inhibitor that inhibits P10721 , enhances engraftment after bone marrow transplantation ( BMT ) in mice . DB01268 diminished hematopoietic progenitor cell numbers , and sunitinib enhanced marrow , peripheral myeloid , and lymphoid engraftment after BMT in Rag1(-/-) mice . DB01268 augmented P19526 engraftment because recipients displayed increased myeloid and lymphoid engraftment and because sunitinib-treated recipients of purified HSCs showed enhanced engraftment of secondary hosts . However , sunitinib preferentially augmented T-cell engraftment with lesser effects on myeloid and P19526 engraftment . Consistent with this , sunitinib preferentially depleted the early thymic progenitor subset in the thymus . DB01268 did not increase engraftment in mice with deficient P10721 signaling , and the pattern of more potent effects on T cell compared with P19526 engraftment observed in sunitinib-treated hosts was also observed after BMT into P10721 (W/Wv) mice . These results implicate P10721 as a critical modulator of thymic niches . We conclude that transient , pharmacologic inhibition of P10721 enhances accessibility of marrow and thymic niches , and provides a novel , noncytotoxic approach to accomplish engraftment after stem cell transplantation . [ Imatinib . DB01268 ] . Imatinib mesilate , which efficiently inhibits P11274 - P00519 ,and P10721 as well as platelet-derived growth factor receptor ( PDGF-R ) kinases , is highly effective for clinical treatment of CML , Ph+ALL , and advanced GIST with good tolerability , respectively . Acquired resistance to the drug,however , becomes an clinically emerging problem with long-standing use . Meanwhile , sunitinib malate,which inhibits three P15692 -Rs and P17948 3 in addition to P10721 as well as PDGF-R , was clinically evaluated in the phase II clinical trials for imatinib-resistant or intolerant GIST , and advanced renal cell carcinoma in Japan . DB01268 is therapeutically effective for both on imatinib-resistant GIST and advanced renal cell carcinoma with modest tolerability , and is now under review for approval in Japan . Effects of preset sequential administrations of sunitinib and everolimus on tumour differentiation in Caki-1 renal cell carcinoma . BACKGROUND : DB01268 ( VEGFR/ P09619 inhibitor ) and everolimus ( P42345 inhibitor ) are both approved for advanced renal cell carcinoma ( RCC ) as first-line and second-line therapy , respectively . In the clinics , sunitinib treatment is limited by the emergence of acquired resistance , leading to a switch to second-line treatment at progression , often based on everolimus . No data have been yet generated on programmed alternating sequential strategies combining alternative use of sunitinib and everolimus before progression . Such strategy is expected to delay the emergence of acquired resistance and improve tumour control . The aim of our study was to assess the changes in tumours induced by three different sequences administration of sunitinib and everolimus . METHODS : In human Caki-1 RCC xenograft model , sunitinib was alternated with everolimus every week , every 2 weeks , or every 3 weeks . Effects on necrosis , hypoxia , angiogenesis , and EMT status were assessed by immunohisochemistry and immunofluorescence . RESULTS : DB01268 and everolimus programmed sequential regimens before progression yielded longer median time to tumour progression than sunitinib and everolimus monotherapies . In each group of treatment , tumour growth control was associated with inhibition of P42345 pathway and changes from a mesenchymal towards an epithelial phenotype , with a decrease in vimentin and an increase in P12830 expression . The sequential combinations of these two agents in a RCC mouse clinical trial induced antiangiogenic effects , leading to tumour necrosis . CONCLUSIONS : In summary , our study showed that alternate sequence of sunitinib and everolimus mitigated the development of mesenchymal phenotype compared with sunitinib as single agent . Stimulation of epithelial repair is a likely mechanism for the action of mifepristone in reducing duration of bleeding in users of progestogen-only contraceptives . Many women using progestogen ( P ) -only contraceptives experience uterine bleeding problems . In clinical trials , a single low dose of mifepristone , given to DB00294 users at the beginning of a bleeding episode reduced the number of bleeding days by approximately 50 % compared with controls . In this study , a single dose of mifepristone was administered to etonogestrel ( P17813 ) -exposed pseudo-pregnant mice , 5 days after artificial decidualization was induced when the endometrium showed signs of bleeding . Control mice received vehicle alone . Mice were culled 12- , 18- , 24- and 48-h post-treatment . In the continued presence of P17813 , a single dose of mifepristone stimulated tissue breakdown followed by very rapid repair : most treated tissues were fully restored to the pre-decidualized state by 48 h post-treatment . During repair , proliferating cells ( Ki67 immunostained ) were localized to a band of cells around the basal area in breaking down tissues and to the repairing luminal epithelium and glands . P06401 -positive cells were largely localized to the basal area of the breaking down tissue in treated mice compared with decidual cells in controls . Oestrogen receptor-positive cells were observed in the repairing luminal epithelium and glands compared with the decidua and the basal region in control tissues . It is concluded that mifepristone treatment stimulates rapid restoration of luminal epithelial integrity : such action may be a key event in reducing the number of bleeding days observed in women using DB00294 who were treated with a single dose of mifepristone . Immunohistochemical expression of P12830 does not distinguish canine cutaneous histiocytoma from other canine round cell tumors . Immunohistochemistry for P12830 ( ECAD ) has been used to distinguish canine cutaneous histiocytoma from other leukocytic neoplasms ( " round cell tumors " ) . To determine the specificity of this test , 5 types of canine cutaneous round cell tumors were evaluated for immunohistochemical expression of ECAD . Tumors of all 5 types had variable cytoplasmic , plasma membrane , and/or paranuclear ECAD expression : All 13 cutaneous histiocytomas were ECAD+ ; all but 1 of 14 mast cell tumors expressed ECAD ; 10 of 12 epitheliotropic lymphomas reacted with P12830 antibody ; of 72 plasmacytomas , 54 were ECAD+ ; and 5 of 5 histiocytic sarcomas were positive . Conclusions based on these results include the following : First , immunoreactivity for ECAD is not limited to leukocytes of cutaneous histiocytoma ; second , antibody to ECAD also labels neoplastic cells in most mast cell tumors , plasmacytomas , cutaneous histiocytic sarcomas , and epitheliotropic lymphomas ; third , although most histiocytomas have membranous ECAD expression , the immunoreactivity varies among round cell tumors and is frequently concurrent in different cellular compartments ; fourth , the distinctively paranuclear ECAD expression pattern in epitheliotropic lymphomas might distinguish them from other round cell tumors ; and , fifth , ECAD should be used with other markers ( eg , MUM1 for plasmacytomas , P10721 for mast cell tumors , CD3 and CD79a for lymphomas ) to distinguish among canine round cell tumors . Differences in transcript levels of ABC transporters between pancreatic adenocarcinoma and nonneoplastic tissues . OBJECTIVES : The aim of this study was to evaluate transcript levels of all 49 human DB00171 -binding cassette transporters ( ABCs ) in one of the most drug-resistant cancers , namely , the pancreatic ductal adenocarcinoma ( PDAC ) . Association of ABCs levels with clinical-pathologic characteristics and P01116 mutation status was followed as well . METHODS : Tumors and adjacent nonneoplastic tissues were obtained from 32 histologically verified PDAC patients . The transcript profile of ABCs was assessed using quantitative real-time polymerase chain reaction with a relative standard curve . P01116 mutations in exon 2 were assessed by high-resolution melting analysis and sequencing . RESULTS : Most ABCs were deregulated in PDAC and 10 ABCs were associated with clinical-pathologic characteristics . P01116 mutations did not change the global expression profile of ABCs . CONCLUSIONS : The expression of ABC transporters was significantly deregulated in PDAC tumors when compared to nonmalignant tissues . The observed up-regulation of P21439 , O95342 , P33527 , O15438 , O15440 , Q5T3U5 , and Q9UNQ0 in tumors may contribute to the generally poor treatment response of PDAC . The up-regulation of O95477 , Q8IZY2 , and P45844 implicates a serious impairment of cellular cholesterol homeostasis in PDAC . On the other hand , the observed down-regulation of Q99758 , O95255 , P13569 , and Q09428 suggests a possible role of stem cells in the development and progression of PDAC . Recent advances in the treatment of gastrointestinal stromal tumors . Constitutively activating mutations in the P10721 and platelet-derived growth factor receptor α ( P16234 ) RTKs play a crucial role in the biology of gastrointestinal stromal tumors ( GISTs ) , and this disease has served as an effective model for targeting gain-of-function kinase mutations in cancer . Imatinib has entered the clinical arena in the last decade and substantially improved the outcome in these formerly untreatable cancers . However , most advanced GISTs responding to imatinib progress within 2-3 years due to heterogeneous subclones harboring a range of imatinib-resistant secondary P10721 mutations . DB01268 , and more recently , regorafenib , have obtained US Food and Drug Administration approval for the treatment of GISTs after imatinib failure , and thus expanded the treatment options in resistant disease . Within this framework , we present an evaluation of current GIST management , emphasizing the most recent advances in the field together with a discussion on future steps to be taken in refractory disease . Functional blockade of platelet-derived growth factor receptor-beta but not of receptor-alpha prevents vascular smooth muscle cell accumulation in fibrous cap lesions in apolipoprotein E-deficient mice . BACKGROUND : The vascular smooth muscle cell ( VSMC ) is the central cell component involved in the fibroproliferative response in atherogenesis . As the lesion advances , VSMCs migrate from the media into the subendothelial space , thereby forming fibrous plaque lesions . Platelet-derived growth factor ( PDGF ) has been known to be a potent chemoattractant and mitogen for SMCs , but the pathophysiological role of the 2 PDGF receptors , receptor-alpha ( P16234 ) and receptor-beta ( P09619 ) in atherogenesis is poorly understood . To clarify this problem , we prepared antagonistic rat monoclonal antibodies , APA5 and APB5 , against murine P16234 and P09619 , respectively . METHODS AND RESULTS : P02649 -deficient mice were fed a high-fat diet containing 0.3 % cholesterol from 6 weeks of age and subjected to injection with 1 mg/d IP of either antibody from 12 to 18 weeks every other day . In the mice injected with APB5 , the aortic atherosclerotic lesion size and the number of intimal VSMCs were reduced by 67 % and 80 % , respectively , compared with the control mice injected with irrelevant rat IgG . In contrast , the mice that received APA5 showed only minimal reduction of lesion size , and a large number of VSMCs were observed in the intima . In the intima of advanced lesions , APB5 immunolabeled VSMCs , whereas APA5 could detect VSMCs mainly in the media . CONCLUSIONS : These results indicate that P09619 plays a significant role in formation of fibrous atherosclerotic lesions and that regulation of the signal transduction through P09619 could affect atherogenesis in mice . Therapeutic effects of sunitinib , one of the anti-angiogenetic drugs , in a murine arthritis . OBJECTIVES : The purpose of this study was to confirm the inhibitory effects of sunitinib , an angiogenesis inhibitor that targets tyrosine kinases of vascular endothelial growth factor receptor ( VEGFR ) family and platelet-derived growth factor receptor ( P09619 ) family , on arthritis in mice with type II collagen-induced arthritis ( CIA ) . METHODS : DB01268 at a concentration of 30 or 60 mg/kg/day was intraperitoneally administered to mice with CIA . We compared the changes in arthritis score over time , pathological score , bone density , and microvascular density in synovial membrane between the vehicle and treatment groups . RESULTS : In the sunitinib-treated groups , the arthritis score decreased in a dose-dependent manner in comparison with that in the vehicle group . Furthermore , improvement in the pathological score , inhibitory tendency of loss in the bone density , and a decrease in the synovial microvascular density were also observed in the sunitinib-treated groups . CONCLUSIONS : DB01268 remarkably inhibited arthritis , particularly synovial angiogenesis in a murine CIA model . This compound may be useful for treating arthritis . DB01268 in malignant melanoma : a treatment option only for P10721 -mutated patients ? DB01268 has previously been reported to be potentially effective in the treatment of malignant melanomas expressing c- P10721 . Here we report on the case of a 77-year-old gentleman affected by a metastatic clear cell carcinoma of the kidney and a metastatic malignant melanoma with liver and lung metastases . Despite the negativity for CD117 and the absence of P10721 amplification or mutations in the melanoma specimen , he achieved a partial response both in the lungs and in the liver while on sunitinib ( 50 mg once/day , 4 weeks on/2 weeks off ) for the treatment of kidney cancer . To our knowledge , this represents the first evidence of sunitinib activity in P10721 wild-type melanoma . Further studies should be performed to confirm these preliminary data . Gastrointestinal stromal tumor . Gastrointestinal stromal tumors ( GISTs ) are the most common mesenchymal tumors of the gastrointestinal tract . These form a distinct category of tumors characterized by oncogenic mutations of the P10721 receptor tyrosine kinase in a majority of patients . P10721 is used not only for diagnosis but also for targeted therapy of GISTs . Imatinib , a tyrosine kinase inhibitor , is widely used in the treatment of advanced and metastatic GISTs and has been recently employed in the neo adjuvant and adjuvant set-up with encouraging results . Certain specific mutations in an exon ( such as in exon 9 ) of the P10721 gene result in GISTs that are relatively unresponsive to the Imatinib treatment . New therapeutic agents like DB01268 have now been approved for the treatment of Imatinib-resistant GIST . This review summarizes the salient features of GIST along with a detailed review of targeted multi-disciplinary approach to the treatment of these special tumors . Pathways targeted by antidiabetes drugs are enriched for multiple genes associated with type 2 diabetes risk . Genome-wide association studies ( GWAS ) have uncovered > 65 common variants associated with type 2 diabetes ( T2D ) ; however , their relevance for drug development is not yet clear . Of note , the first two T2D-associated loci ( P37231 and Q14654 / Q09428 ) encode known targets of antidiabetes medications . We therefore tested whether other genes/pathways targeted by antidiabetes drugs are associated with T2D . We compiled a list of 102 genes in pathways targeted by marketed antidiabetic medications and applied Gene Set Enrichment Analysis ( MAGENTA [ Meta-Analysis Gene-set Enrichment of variaNT Associations ] ) to this gene set , using available GWAS meta-analyses for T2D and seven quantitative glycemic traits . We detected a strong enrichment of drug target genes associated with T2D ( P = 2 × 10(-5) ; 14 potential new associations ) , primarily driven by insulin and thiazolidinedione ( TZD ) targets , which was replicated in an independent meta-analysis ( Metabochip ) . The glycemic traits yielded no enrichment . The T2D enrichment signal was largely due to multiple genes of modest effects ( P = 4 × 10(-4) , after removing known loci ) , highlighting new associations for follow-up ( P33121 , P19838 , P11168 , incretin targets ) . Furthermore , we found that TZD targets were enriched for LDL cholesterol associations , illustrating the utility of this approach in identifying potential side effects . These results highlight the potential biomedical relevance of genes revealed by GWAS and may provide new avenues for tailored therapy and T2D treatment design . Raddeanin A , a triterpenoid saponin isolated from Anemone raddeana , suppresses the angiogenesis and growth of human colorectal tumor by inhibiting P35968 signaling . Raddeanin A ( RA ) is an active triterpenoid saponin from a traditional Chinese medicinal herb , Anemone raddeana Regel . It was previously reported that RA possessed attractive antitumor activity through inhibiting proliferation and inducing apoptosis of multiple cancer cells . However , whether RA can inhibit angiogenesis , an essential step in cancer development , remains unknown . In this study , we found that RA could significantly inhibit human umbilical vein endothelial cell ( HUVEC ) proliferation , motility , migration , and tube formation . RA also dramatically reduced angiogenesis in chick embryo chorioallantoic membrane ( P62158 ) , restrained the trunk angiogenesis in zebrafish , and suppressed angiogenesis and growth of human HCT-15 colorectal cancer xenograft in mice . Western blot assay showed that RA suppressed P15692 -induced phosphorylation of P35968 and its downstream protein kinases including PLCγ1 , O60674 , Q05397 , Src , and Akt . Molecular docking simulation indicated that RA formed hydrogen bonds and hydrophobic interactions within the DB00171 binding pocket of P35968 kinase domain . Our study firstly provides the evidence that RA has high antiangiogenic potency and explores its molecular basis , demonstrating that RA is a potential agent or lead candidate for antiangiogenic cancer therapy . Circulating cytokines and monocyte subpopulations as biomarkers of outcome and biological activity in sunitinib-treated patients with advanced neuroendocrine tumours . BACKGROUND : DB01268 is approved worldwide for treatment of advanced pancreatic neuroendocrine tumours ( pNET ) , but no validated markers exist to predict response . This analysis explored biomarkers associated with sunitinib activity and clinical benefit in patients with pNET and carcinoid tumours in a phase II study . METHODS : Plasma was assessed for vascular endothelial growth factor ( P15692 ) -A , soluble P15692 receptor ( sVEGFR ) -2 , sVEGFR-3 , interleukin ( IL ) -8 ( n=105 ) , and stromal cell-derived factor ( SDF ) -1α ( n=28 ) . Pre-treatment levels were compared between tumour types and correlated with response , progression-free ( PFS ) , and overall survival ( OS ) . Changes in circulating myelomonocytic and endothelial cells were also analysed . RESULTS : Stromal cell-derived factor-1α and sVEGFR-2 levels were higher in pNET than in carcinoid ( P=0.003 and 0.041 , respectively ) . High ( above-median ) baseline SDF-1α was associated with worse PFS , OS , and response in pNET , and high sVEGFR-2 with longer OS ( P⩽0.05 ) . For carcinoid , high P10145 , sVEGFR-3 , and SDF-1α were associated with shorter PFS and OS , and high P10145 and SDF-1α with worse response ( P⩽0.05 ) . Among circulating cell types , monocytes showed the largest on-treatment decrease , particularly P08571 + monocytes co-expressing P17948 or P61073 . CONCLUSIONS : P10145 , sVEGFR-3 , and SDF-1α were identified as predictors of sunitinib clinical outcome . Putative pro-tumorigenic P61073 + and P17948 + monocytes represent novel candidate markers and biologically relevant targets explaining the activity of sunitinib . Tyrosine kinase blockers : new hope for successful cancer therapy . Tyrosine kinases ( TKs ) are attractive targets for cancer therapy , as quite often their abnormal signaling has been linked with tumor development and growth . Constitutive activated TKs stimulate multiple signaling pathways responsible for DNA repair , apoptosis , and cell proliferation . During the last few years , thorough analysis of the mechanism underlying tyrosine kinase 's activity led to novel cancer therapy using TKs blockers . These drugs are remarkably effective in the treatment of various human tumors including head and neck , gastric , prostate and breast cancer and leukemias . The most successful example of kinase blockers is Imatinib ( Imatinib mesylate , Gleevec , STI571 ) , the inhibitor of Bcr/Abl oncoprotein , which has become a first-line therapy for chronic myelogenous leukemia . The introduction of STI571 for the treatment of leukemia in clinical oncology has had a dramatic impact on how this disease is currently managed . Others kinase inhibitors used recently in cancer therapy include Dasatinib ( BMS-354825 ) specific for P00519 non-receptor cytoplasmic kinase , Gefitinib ( DB00317 ) , Erlotinib ( DB00530 , Tarceva ) and DB01268 ( SU 11248 , Sutent ) specific for P15692 receptor kinase , AMN107 ( DB04868 ) and INNO-406 ( NS-187 ) specific for c- P10721 kinase . The following TK blockers for treatment of various human tumors are in clinical development : DB01259 ( DB01259 ditosylate , DB01259 , GW-572016 ) , Canertinib ( DB05424 ) , DB05294 ( DB05294 ) , DB04879 ( PTK787/ZK 222584 ) , DB00398 ( Bay 43-9006 , Nexavar ) , and Leflunomide ( SU101 , DB01097 ) . Herein , we discuss the chemistry , biological activity and clinical potential of new drugs with tyrosine kinase blockers for cancer treatment . New paradigms in gastrointestinal stromal tumour management . BACKGROUND : Targeted agents have improved the prognosis for patients with advanced gastrointestinal stromal tumours ( GISTs ) . Many patients exhibit intolerance or resistance to first-line therapy with imatinib mesylate . DB01268 malate is approved multinationally for the treatment of advanced imatinib-refractory GIST . DESIGN : This article reviews responses to imatinib and sunitinib reported in clinical trials in advanced GIST and discusses the effect of mutational status on treatment responses ; therapeutic developments in GIST treatment are also reviewed . RESULTS : Imatinib 400 mg/day has shown efficacy for first-line treatment of advanced GIST , particularly in patients with P10721 exon 11 mutations . DB01268 50 mg/day ( Schedule 4/2 ) has demonstrated effectiveness and tolerability in imatinib-refractory GIST , including patients who would be excluded from clinical trials . DB01268 is associated with longer median overall survival in patients with primary P10721 exon 9 mutations and wild-type GIST compared with P10721 exon 11 mutations in a retrospective study . Ongoing studies , including imatinib in the adjuvant setting and the use of targeted agents in sequence or in combination , will further refine the therapeutic pathway for advanced GIST . CONCLUSIONS : The availability of targeted therapies and greater knowledge of the effect of mutational status on patient responses will assist in optimising outcomes in advanced GIST . Expression of vascular endothelial growth factor ( P15692 ) and its receptors P17948 and P35968 in primary and recurrent WHO grade III meningiomas . AIMS : WHO grade III meningiomas are malignant neoplasms for which new and more targeted treatment strategies are urgently needed . Although clinical trials investigating anti-angiogenic vascular endothelial growth factor ( P15692 ) targeted therapies are currently recruiting , knowledge about the expression of P15692 and P15692 receptors remains to be determined . METHODS : We investigated the expression of P15692 and its receptors P17948 and P35968 in 32 WHO grade III meningioma samples by immunohistochemistry . Furthermore , we performed in-situ hybridisation for P15692 . RESULTS : We found low P15692 expression in tumor and endothelial cells . Highest P15692 expression levels were seen in peri-necrotic tumor cells potentially suffering from hypoxia . P17948 and 2 were virtually absent on tumor cells , although endothelial cells displayed significantly higher levels reaching stronger expression for P35968 than P17948 . CONCLUSIONS : Our findings showing constant expression levels of P35968 in endothelial cells serve as a first indication that the use of small tyrosine kinase inhibitors such as DB01268 directly targeting the P15692 -receptors might be worth testing , also in the clinical context in cases of therapy-refractory meningiomas . Further investigations are needed to study the response to drugs targeting the P15692 pathway in relation to the expression profile of P15692 and its receptors in high grade meningiomas . Complementary and alternative medicine use and quality of life in patients with primary brain tumors . This study explored the use of complementary and alternative medicine ( P62158 ) approaches and their relationship with demographic and disease characteristics and quality of life ( QOL ) in the primary brain tumor ( P10721 ) population . One hundred one P10721 patients were enrolled in this study . The results showed that 34 % of patients reported using P62158 . Forty-one percent reported using more than one type of P62158 . The average cost of each P62158 used per month was 69 dollars , with 20 % of patients spending more than 100 dollars per month . The majority ( 74 % ) reported that their physicians were unaware of their use of P62158 . Data analysis found a higher performance status to be the only factor significantly related to use of P62158 therapy ( P < 0.005 ) . There was no difference in patient report of QOL between users and nonusers of P62158 therapies . The high number of patients who do not report P62158 use has potential implications for evaluation of symptoms and response to therapy in this population . This may be especially relevant in those patients with higher functional status participating in clinical trials . DB01268 ( SUTENT , SU11248 ) suppresses tumor growth and induces apoptosis in xenograft models of human hepatocellular carcinoma . Hepatocellular carcinoma ( HCC ) is the fifth most common and third deadliest primary neoplasm . Since HCC is a particularly vascular solid tumor , we determined the antitumor and antiangiogenic activities of sunitinib malate , a potent inhibitor of two receptors involved in angiogenesis - vascular endothelial growth factor receptor ( VEGFR ) and platelet-derived growth factor receptor ( P09619 ) . In the present study , we reported that treatment of HepG2 and SK-Hep-1 cells with sunitinib led to growth inhibition and apoptosis in a dose-dependent fashion . DB01268 inhibited phosphorylation of P35968 at Tyr951 and P09619 at Tyr1021 both in vitro and in vivo . DB01268 also suppressed tumor growth of five patient-derived xenografts . DB01268 -induced tumor growth inhibition was associated with increased apoptosis , reduced microvessel density and inhibition of cell proliferation . This study provides a strong rationale for further clinical investigation of sunitinib in patients with hepatocellular carcinoma . Blocking autocrine P15692 signaling by sunitinib , an anti-cancer drug , promotes embryonic stem cell self-renewal and somatic cell reprogramming . Maintaining the self-renewal of embryonic stem cells ( ESCs ) could be achieved by activating the extrinsic signaling , i.e. , the use of leukemia inhibitory factor ( P15018 ) , or blocking the intrinsic differentiation pathways , i.e. , the use of GSK3 and MEK inhibitors ( 2i ) . Here we found that even in medium supplemented with P15018 , mESCs still tend to differentiate toward meso-endoderm lineages after long-term culture and the culture spontaneously secretes vascular endothelial growth factors ( VEGFs ) . Blocking P15692 signaling with sunitinib , an anti-cancer drug and a receptor tyrosine kinase ( RTK ) inhibitor mainly targeting P15692 receptors ( VEGFRs ) , is capable of maintaining the mESCs in the undifferentiated state without the need for feeder cells or P15018 . DB01268 facilitates the derivation of mESCs from blastocysts , and the mESCs maintained in sunitinib-containing medium remain pluripotent and are able to contribute to chimeric mice . DB01268 also promotes iPSC generation from MEFs with only Oct4 . Knocking down P35968 or blocking it with neutralizing antibody mimicks the effect of sunitinib , indicating that blocking P15692 /VEGFR signaling is indeed beneficial to the self-renewal of mESCs . We also found that hypoxia-inducible factor alpha ( HIF1α ) and endoplasmic reticulum ( ER ) stress are involved in the production of P15692 in mESCs . Blocking both pathways inhibits the expression of P15692 and prevents spontaneous differentiation of mESCs . Interestingly , P15018 may also exert its effect by downregulating HIF1α and ER stress pathways and subsequent P15692 expression . These results indicate the existence of an intrinsic differentiation pathway in mESCs by activating the autocrine P15692 signaling . Blocking P15692 signaling with sunitinib or other small molecules help to maintain the mESCs in the ground state of pluripotency . P01308 / P05019 signaling pathways enhances tumor cell invasion through bisecting GlcNAc N-glycans modulation . an interplay with P12830 . Changes in glycosylation are considered a hallmark of cancer , and one of the key targets of glycosylation modifications is P12830 . We and others have previously demonstrated that P12830 has a role in the regulation of bisecting GlcNAc N-glycans expression , remaining to be determined the P12830 -dependent signaling pathway involved in this N-glycans expression regulation . In this study , we analysed the impact of P12830 expression in the activation profile of receptor tyrosine kinases such as insulin receptor ( IR ) and P08069 ( IGF-IR ) . We demonstrated that exogenous P12830 expression inhibits IR , IGF-IR and P29323 1/2 phosphorylation . Stimulation with insulin and P05019 in MDA-MD-435 cancer cells overexpressing P12830 induces a decrease of bisecting GlcNAc N-glycans that was accompanied with alterations on P12830 cellular localization . Concomitantly , IR/IGF-IR signaling activation induced a mesenchymal-like phenotype of cancer cells together with an increased tumor cell invasion capability . Altogether , these results demonstrate an interplay between P12830 and IR/IGF-IR signaling as major networking players in the regulation of bisecting N-glycans expression , with important effects in the modulation of epithelial characteristics and tumor cell invasion . Here we provide new insights into the role that P01308 / P05019 signaling play during cancer progression through glycosylation modifications . A new cell culture-based assay quantifies vitamin K 2,3-epoxide reductase complex subunit 1 function and reveals warfarin resistance phenotypes not shown by the dithiothreitol-driven Q9BQB6 assay . BACKGROUND : DB00682 directly inhibits the vitamin K 2,3-epoxide reductase complex subunit 1 ( Q9BQB6 ) enzyme to effect anticoagulation . Q9BQB6 function has historically been assessed in vitro using a dithiothreitol ( DTT ) -driven vitamin K 2,3-epoxide reductase ( Q9BQB6 ) assay . DB00682 inhibits wild-type Q9BQB6 function by the DTT- Q9BQB6 assay . However , Q9BQB6 variants with warfarin resistance-associated missense mutations often show low Q9BQB6 activities and warfarin sensitivity instead of resistance . OBJECTIVES : A cell culture-based , indirect Q9BQB6 assay was developed and characterized that accurately reports warfarin sensitivity or resistance for wild-type and variant Q9BQB6 proteins . METHODS : Human coagulation factor (F)IX and Q9BQB6 variants were coexpressed in P29320 293T cells under standardized conditions at various warfarin concentrations . Secreted FIX activity served as surrogate marker to report wild-type and variant Q9BQB6 inhibition by warfarin . RESULTS AND CONCLUSIONS : DB00682 dose-response curves fit to the secreted FIX activity data for coexpressed hVKORC1 wild-type , Val29Leu , Val45Ala and Leu128Arg variants . The corresponding calculated IC50 values were 24.7 , 136.4 , 152.0 and 1226.4 nm , respectively . Basal activities in the absence of warfarin for all Q9BQB6 variants were similar to that of wild-type Q9BQB6 . Ranked IC50 values from the cell culture-based assay accurately reflect elevated warfarin dosages for patients with Q9BQB6 missense mutation-associated warfarin resistance . Implantation of P15692 transfected preadipocytes improves vascularization of fibrin implants on the cylinder chorioallantoic membrane ( P62158 ) model . The successful substitution or augmentation of soft tissues by implantation of three dimensional cell constructs , consisting of human preadipocytes and fibrin glue as a carrier matrix , requires a rapid and homogeneous vascularization of the whole implant in order to provide a sufficient blood supply of centrally situated cells . Previous investigations have shown that under in vivo conditions primary human preadipocytes induce vascularization of fibrin matrices by secretion of several growth factors , such as P15692 and P09038 . The current study investigates whether vascularization of implants can be improved by transplantation of preadipocytes following transfection with a P15692 -vector . Transfection was performed by electroporation with an pCMX-GFP and pCMX-VEGF165 vector . Transfection efficiency ( GFP expression ) and P15692 expression were determined in vitro by FACS analysis and P15692 immunoassay , respectively . In vivo investigations to determine the vascularization of the implants were performed on the cylinder chorioallantoic membrane ( P62158 ) . Four million P15692 transfected cells were transferred within a fibrin matrix onto the P62158 on the 7(th) day of incubation and after 8 days the vascularization of the implant was histologically examined and evaluated by means of a computer-assisted image analysis program . Transfection of preadipocytes with the GFP vector by electroporation yielded transfection efficiencies between 12 % and 41 % of surviving cells . Results of the P15692 immunoassay demonstrated that P15692 expression was significantly higher following transfection . Investigations on the P62158 outlined a significantly higher rate of vascularization in the transfected vs. control population . Our investigations demonstrate that primary human preadipocytes can be successfully transfected by electroporation with a P15692 vector . The enhanced P15692 expression on transfected cells results in an increase of vascularization of the cell constructs on the P62158 . Molecular target modulation , imaging , and clinical evaluation of gastrointestinal stromal tumor patients treated with sunitinib malate after imatinib failure . PURPOSE : To evaluate sunitinib activity and potential cellular and molecular correlates in gastrointestinal stromal tumor ( GIST ) patients after imatinib failure , in addition to assessing the safety and pharmacokinetics ( PK ) of different dose schedules . EXPERIMENTAL DESIGN : In this open-label , dose-ranging , phase I/II study , 97 patients with metastatic imatinib-resistant/intolerant GIST received sunitinib at doses of 25 , 50 , or 75 mg/d on one of three schedules . Serial tumor imaging was done using computed tomography and [18F]fluoro-2-deoxy-d-glucose positron emission tomography scanning . PK and cell proliferation and P10721 phosphorylation status in tumor biopsies were also analyzed . RESULTS : Clinical benefit was observed in 52 patients ( 54 % : 7 objective partial responses , 45 stable disease > or =6 months ) . Decreased tumor glycolytic activity was shown in most patients within 7 days of starting sunitinib using [18F]fluoro-2-deoxy-d-glucose positron emission tomography . DB01268 treatment was associated with reduced tumor cell proliferation by > 25 % in 52 % of cases analyzed and reduced levels of phospho- P10721 in tumor biopsies ( indicating target modulation ) . The recommended dose schedule was 50 mg/d for 4 weeks followed by 2 weeks off treatment . On the 50-mg dose across all schedules , 79 % of PK-evaluable patients achieved total drug trough concentrations above the target concentration ( 50 ng/mL ) within 14 days of dosing . In addition , adverse events were generally mild to moderate in severity . CONCLUSION : Cellular and molecular analyses showed that sunitinib clinical activity is associated with inhibition of P10721 in GIST following imatinib failure , illustrating the rational approach used to develop a therapy aimed at the underlying oncogenic signaling pathway aberrancy . DB01268 in advanced alveolar soft part sarcoma : evidence of a direct antitumor effect . BACKGROUND : The purpose of this study was to confirm sunitinib activity in alveolar soft part sarcoma ( ASPS ) and to report on new insights into the molecular bases thereof . PATIENTS AND METHODS : From July 2007 , nine patients with progressive metastatic ASPS received sunitinib 37.5 mg/day , within a named use program . Cryopreserved material was available for five naive patients , among whom three received sunitinib . Immunofluorescence ( IF ) /confocal microscopy , biochemical , and molecular/cytogenetic analyses were carried out , complemented by antiproliferative and activation assays in a short-term culture derived from one case . RESULTS : All patients were eligible for response . Best RECIST response was partial response in five cases , stable disease in three , and progression in one . The median progression-free survival was 17 months . Positron emission tomography results were consistent . Two cases of interval progressions were recorded . Antiproliferative assays and biochemistry on short-term culture showed that sunitinib is able to markedly impair ASPS cells growth and switch-off P09619 . IF/confocal microscopy demonstrated coexpression and physical association between P09619 /vascular endothelial growth factor receptor 2 ( P35968 ) and P07949 / P35968 in ASPS cells , which was validated by biochemistry . P09619 , P07949 , and MET ligand-dependent activation was confirmed . CONCLUSIONS : We confirm the clinical efficacy of sunitinib in ASPS , mediated by P09619 , P35968 , and P07949 , which are all expressed in tumor cells . A direct antitumor effect was shown in a short-term cell culture . Gender difference in the activity but not expression of estrogen receptors alpha and beta in human lung adenocarcinoma cells . The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer . We evaluated estrogen receptor ( ER ) alpha and beta expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts . Q8N1N2 -length ERalpha and ERbeta proteins were expressed in all cell lines with higher ERbeta than ERalpha . Although estradiol ( E(2) ) binding was similar , E(2) stimulated proliferation only in cells from females , and this response was inhibited by anti-estrogens 4-hydroxytamoxifen ( DB04468 ) and DB00947 . In contrast , E(2) did not stimulate replication of lung adenocarcinoma cells from males and DB04468 or ICI did not block cell proliferation . Similarly , transcription of an estrogen response element-driven reporter gene was stimulated by E(2) in lung adenocarcinoma cells from females , but not males . P06401 ( PR ) expression was increased by E(2) in two out of five adenocarcinoma cell lines from females , but none from males . E(2) decreased P12830 protein expression in some of the cell lines from females , as it did in MCF-7 breast cancer cells , but not in the cell lines from males . Thus , ERalpha and ERbeta expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells . On the other hand , coactivator Q15648 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells . Q15648 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males . Effects of ellagic Acid on angiogenic factors in prostate cancer cells . BACKGROUND : Several natural antioxidants , including ellagic acid ( EA ) , have been reported to have chemotherapeutic activity in vivo and in vitro settings . Cytochrome P450 ( CYP ) activity and synthesis of both epoxyeicosatrienoic acids ( EETs ) and 20-hydroxy-5,8,11,14-eicosatetraenoic acid ( 20-HETE ) , together with vascular endothelial growth factor ( P15692 ) and heme oxygenase system ( HO ) have emerged as important modulators of tumor growth and metastasis . METHODS : The anti-angiogenic effects of EA were investigated in the human prostatic cancer cell line LnCap . P09601 , P30519 , P51589 and soluble epoxyde hydrolase ( sEH ) expressions were evaluated by western blotting . Levels of P15692 and osteoprotegerin ( O00300 ) were determined in the culture supernatant using an ELISA assay , while CYP mRNAs were determined by qRT-PCR . RESULTS : EA treatment induced a significant decrease ( p < 0.05 ) in P09601 , P30519 and P51589 expression , and in P15692 and O00300 levels . Similarly P51589 , P78329 and CYPA22 mRNAs were significantly ( p < 0.05 ) down-regulated by EA treatment . The decrease in P51589 mRNA was associated with an increase in sEH expression . CONCLUSIONS : RESULTS reported in the present study highlighted the ability of EA to modulate a new pathway , in addition to anti-proliferative and pro-differentiation properties , via a mechanism that involves a decrease in eicosanoid synthesis and a down-regulation of the HO system in prostate cancer . EV71 induces P35354 expression via c-Src/ P09619 /PI3K/Akt/ Q8NFH3 / Q8TCB0 MAPK/AP-1 and NF-kappaB in rat brain astrocytes . Enterovirus 71 ( EV71 ) induces the expression of cyclooxgenase ( P36551 ) -2 served as a major neurotoxic factor in CNS injury . However , the mechanisms underlying EV71-initiated intracellular signaling pathways leading to P35354 expression remain unknown . Therefore , we investigated the mechanisms underlying EV71-induced P35354 expression and prostaglandin E(2) ( PGE(2) ) production in rat brain astrocytes ( RBA ) -1 , determined by Western blotting , RT-PCR , and promoter assay . Here , we reported that EV71-induced P35354 expression and PGE(2) production were attenuated by pretreatment with the inhibitors of c-Src ( P50391 ) , P09619 ( AG1296 ) , PI3K ( Wortmannin ) , Q02750 /2 ( PD98059 ) , NF-kappaB ( helenalin ) , and AP-1 ( Tanshinone ) and transfection with shRNA or siRNA of c-Src , P09619 , p85 , c-Jun , c-Fos , P27361 , or P28482 . We further observed that EV71-induced activation of Akt and Q8NFH3 / Q8TCB0 MAPK were mediated via c-Src and P09619 . Pretreatment with P50391 attenuated EV71-stimulated phosphorylation of Src , P09619 , Akt , and Q8NFH3 / Q8TCB0 MAPK . Inhibition of PI3K by Wortmannin attenuated EV71-induced Akt and Q8NFH3 / Q8TCB0 MAPK phosphorylation , but had no effect on P09619 phosphorylation , suggesting that P09619 is an upstream and Q8NFH3 / Q8TCB0 MAPK is a downstream component of PI3K/Akt in these responses . EV71-stimulated NF-kappaB translocation from the cytoplasm to the nucleus , P25963 degradation and NF-kappaB promoter activity were attenuated by pretreatment with helenalin , but not AG1296 , Wortmannin , and PD98059 . EV71-induced c-Jun mRNA expression was attenuated by pretreatment with PD98059 , AG1296 , or Wortmannin . These results demonstrate that in RBA-1 cells , EV71-induced P35354 expression associated with PGE(2) production is mediated through activation of c-Src/ P09619 /PI3K/Akt/ Q8NFH3 / Q8TCB0 MAPK to initiate the expression of AP-1 . DB01268 synergizes the antitumor effect of cisplatin via modulation of P07992 expression in models of gastric cancer . We evaluated the effects of sunitinib monotherapy and in combination with cisplatin in human gastric cancer cell lines . DB01268 showed antiproliferative effect in gastric cancer cells line with high P16234 expression . Knockdown of P16234 showed that sunitinib sensitivity was correlated with the basal expression of P16234 . Synergistic growth inhibitory activity in combination with cisplatin was identified . We further explored how sunitinib potentiated the activity of cisplatin . We found that sunitinib treatment resulted in the down-regulation of P07992 expression via the modulation of P16234 expression in gastric cancer cells . The effect was verified via SNU484 xenograft model . Our data support the rationale of clinical trial using sunitinib in combination of cisplatin in gastric cancer . DB01268 mesylate inhibits proliferation of human colonic stromal fibroblasts in vitro and in vivo . OBJECTIVE : Cancer stromal fibroblasts are important members of the cancer microenvironment . In this study , we determined the effect of sunitinib , a small molecule tyrosine kinase inhibitor , on the primary human colonic fibroblasts . METHODS : Cell cycle analysis and cell proliferation assays were performed to evaluate the inhibitory effect of sunitinib in vitro . Western-blot analysis was performed to evaluate variations in the levels of phosphorylated platelet-derived growth factor receptor β ( P09619 -β ) , Akt , and P29323 proteins . Co-injection of SW620 cells and colonic fibroblasts in nude mice was employed to test anti-growth efficacy in vivo . RESULTS : DB01268 was found to effectively inhibit the growth of primary colonic fibroblasts . Low-dose sunitinib blocked the DB00102 -induced cell proliferation and P09619 -β signaling . Co-injection of SW620 cells and colonic fibroblasts in nude mice generated greater tumor volumes than single injection of SW620 cells . DB01268 treatment inhibited the SW620 cell+colonic fibroblast tumor growth more effectively than treatment of 5-fluorouracil . CONCLUSIONS : DB01268 mesylate inhibited the proliferation of primary human colonic fibroblasts through target-inhibited P09619 signaling in vitro and in vivo . Differential regulation of sunitinib targets predicts its tumor-type-specific effect on endothelial and/or tumor cell apoptosis . PURPOSE : DB01268 is an inhibitor of tyrosine-kinase receptors , and no biomarker predictive of sunitinib response is available . The purpose of this preclinical study was to show whether sunitinib molecular targets could be used as biomarkers to assess tumor response to sunitinib in human cancer cell line xenografts of three different tumor types . METHODS : Using mice xenografted with liver , breast and renal carcinoma cell lines , we sequentially analyzed the effect of 7-day sunitinib treatment on tumor and vascular compartments . RESULTS : In all xenografts , microvessel damage occurred from Day 1 . Tumor damage also occurred in liver , breast , but not in renal xenografts . Using specific human and mouse probes for genes encoding sunitinib targets , we showed a significant relation between apoptotic tumor cell numbers and human PDGFRΒ and P07949 mRNA expression in liver cancer and to human P35968 expression in breast cancer xenografts . In contrast , in renal cancer xenografts , vascular effect evaluated by measuring endothelial cell apoptosis was related to mouse Vegfr1 , Vegfr2 and Vegfa-164 expression . CONCLUSION : This study identifies sunitinib vascular and tumor effects according to different tumor types and shows that sunitinib molecular targets used as biomarkers enable assessment of therapeutic response .