ABSTRACT
Urothelial carcinoma (UC) carcinogenesis has been hypothesized to occur through epigenetic repression of tumor-suppressor genes (TSGs). By quantitative real-time polymerase chain reaction array, we found that one potential TSG, angiopoietin-like 4 (ANGPTL4), was expressed at very low levels in all bladder cancer cell lines we examined. Previous studies had demonstrated that ANGPTL4 is highly expressed in some cancers, but downregulated, by DNA methylation, in others. Consequently, owing to these seemingly conflicting functions in distinct cancers, the precise role of ANGPTL4 in the etiology of UC remains unclear. In this study, using methylation-specific PCR and bisulfite pyrosequencing, we show that ANGPTL4 is transcriptionally repressed by DNA methylation in UC cell lines and primary tumor samples, as compared with adjacent noncancerous bladder epithelium. Functional studies further demonstrated that ectopic expression of ANGPTL4 potently suppressed UC cell proliferation, monolayer colony formation in vitro, and invasion, migration, and xenograft formation in vivo. Surprisingly, circulating ANGPTL4 was significantly higher in plasma samples from UC patients than normal control, suggesting it might be secreted from other cell types. Interestingly, our data also indicated that exogenous cANGPTL4 could promote cell proliferation and cell migration via activation of signaling through the Erk/focal adhesion kinase axis. We further confirmed that mouse xenograft tumor growth could be promoted by administration of exogenous cANGPTL4. Finally, immunohistochemistry demonstrated that ANGPTL4 was downregulated in tumor cells but overexpressed in tumor adjacent stromal tissues of muscle-invasive UC tissue samples. In conclusion, our data support dual roles for ANGPTL4 in UC progression, either as a tumor suppressor or oncogene, in response to microenvironmental context.
Subject(s)
Angiopoietin-Like Protein 4/genetics , Carcinoma, Transitional Cell/genetics , Epigenesis, Genetic/genetics , Gene Expression Regulation, Neoplastic/genetics , Tumor Microenvironment , Urinary Bladder Neoplasms/genetics , Aged , Aged, 80 and over , Angiopoietin-Like Protein 4/blood , Angiopoietin-Like Protein 4/metabolism , Animals , Carcinogenesis/genetics , Carcinoma, Transitional Cell/pathology , Carcinoma, Transitional Cell/surgery , Case-Control Studies , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Cystectomy , DNA Methylation/genetics , Down-Regulation , Female , Genes, Tumor Suppressor , Humans , Male , Mice , Mice, Inbred NOD , Mice, SCID , Middle Aged , Oncogenes/genetics , Promoter Regions, Genetic/genetics , Signal Transduction , Urinary Bladder/pathology , Urinary Bladder/surgery , Urinary Bladder Neoplasms/pathology , Urothelium/pathology , Xenograft Model Antitumor AssaysABSTRACT
We report the first genome-wide association study in 1000 bipolar I patients and 1000 controls, with a replication of the top hits in another 409 cases and 1000 controls in the Han Chinese population. Four regions with most strongly associated single-nucleotide polymorphisms (SNPs) were detected, of which three were not found in previous GWA studies in the Caucasian populations. Among them, SNPs close to specificity protein 8 (SP8) and ST8 α-N-acetyl- neuraminide α-2,8-sialyltransferase (ST8SIA2) are associated with Bipolar I, with P-values of 4.87 × 10(-7) (rs2709736) and 6.05 × 10(-6) (rs8040009), respectively. We have also identified SNPs in potassium channel tetramerization domain containing 12 gene (KCTD12) (rs2073831, P=9.74 × 10(-6)) and in CACNB2 (Calcium channel, voltage-dependent, ß-2 subunit) gene (rs11013860, P=5.15 × 10(-5)), One SNP nearby the rs1938526 SNP of ANK3 gene and another SNP nearby the SNP rs11720452 in chromosome 3 reported in previous GWA studies also showed suggestive association in this study (P=6.55 × 10(-5) and P=1.48 × 10(-5), respectively). This may suggest that there are common and population-specific susceptibility genes for bipolar I disorder.
Subject(s)
Bipolar Disorder/ethnology , Bipolar Disorder/genetics , Genetic Predisposition to Disease , Genome-Wide Association Study , Polymorphism, Single Nucleotide/genetics , Ankyrins/genetics , Asian People/ethnology , Asian People/genetics , Bipolar Disorder/epidemiology , Calcium Channels, L-Type/genetics , DNA-Binding Proteins/genetics , Female , Genotype , Humans , Male , Odds Ratio , Phenotype , Proteins/genetics , Reproducibility of Results , Sialyltransferases/genetics , Transcription Factors/geneticsABSTRACT
An expedient liquid-phase synthesis for construction of the diverse benzimidazole libraries is described. Nucleophilic aryl substitution of poly(ethylene glycol)-supported 4-fluoro-3-nitrobenzoic acid 3 with several primary amines under basic conditions, followed by Zn/NH(4)Cl mediated nitro group reduction, gave the PEG bound diamines 5. Subsequent cyclization of immobilized o-phenylenediamine 5 using thiocarbonyldiimidazole (TCD) or thiophosgene in dichloromethane furnished benzimidazole-2-thiones 6. Treatment of 6 with alkyl halides and benzylic halides in the presence of triethylamine provided 1-substituted-2-alkylthio-5-carbamoylbenzimidazoles on the support. The desired products 8 were severed from the PEG under mild conditions in high yield and high purity.
Subject(s)
Benzimidazoles/chemical synthesis , Benzimidazoles/chemistry , Drug Design , Indicators and Reagents , Models, Molecular , Molecular Conformation , Structure-Activity RelationshipSubject(s)
Cholesterol/blood , Coronary Disease/epidemiology , Heart Diseases/epidemiology , Adult , China , Coronary Disease/etiology , Ethnicity , Female , Heart Diseases/etiology , Humans , Male , Middle AgedABSTRACT
Pacemaker stimulation of the left ventricle is expected to yield a pattern of complete right bundle branch block, however, unusual patterns have been reported. The present report is a case of an unusual electrocardiographic pattern from left ventricular endocardial pacing in a patient with an atrial septal defect and acute myocarditis complicated by complete degree atrioventricular block.