Project description:Human livers biopsies from HCV+ patients and healthy donors were collected . RNA and Protein were extract and The Human Adipogenesis RT² Profiler™ PCR Array was used to compare the gene expression in the two group. At the same time PBMCs were collected and the same array were used to compare the adipogenesis expression in the two system.
Project description:Human livers biopsies from HCV+ patients and healthy donors were collected . RNA and Protein were extract and The Human Adipogenesis RT² Profiler™ PCR Array was used to compare the gene expression in the two group. At the same time PBMCs were collected and the same array were used to compare the adipogenesis expression in the two system.
Project description:Introduction: Mechanisms that contribute to the pathogenesis of liver damage caused by hepatitis C virus (HCV) are not fully understood. Our previous work on liver biopsies from chronic HCV patients has shown modulation of the expression of certain cell cycle proteins indicating HCV-induced modifications of cell cycle events. We therefore hypothesize that HCV infection disrupts normal regulation of cell cycle that contributes to disease progression. Objective: To identify molecular disruptions during the course of HCV-associated disease progression, using liver biopsy specimens of chronic hepatitis C patients. Methods: Liver biopsy samples classified on histological basis as early (fibrosis stage 0-1) or advanced (fibrosis stage 3-4) disease stage were studied using oligonucleotide array ( HG U133 Plus 2.0, Affymetrix GeneChip™ System). For comparison, liver specimens from patients with non-viral hepatitis were also analyzed by microarray. Expression data was analyzed using Genespring (GX 7.2) and Ingenuity Pathway analysis (3.0). The differential expression of selected cell cycle genes (cyclin D2, KPNA2, HERC5 and Bcl-2) identified after microarray analysis was confirmed by quantitative real-time RT-PCR. Results: Microarray analysis revealed two-fold or greater transcriptional change in 792 genes of the total 38,500 known human genes in HCV-advance disease stage (HCV-A) as compared to HCV-early disease stage (HCV-E). Most of the genes have a defined role in immune response, extracellular matrix and cell cycle and apoptosis. Keywords: HCV-advance disease state
Project description:In this study, Solexa deep sequencing technology was used for high-throughput analysis of miRNAs in a small RNA library isolated from serum sample of HCV-related fibrosis and control healthy. In total, 41 miRNAs were dysregulated (30 upregulated and 11 downregulated) in the patients with chronic HCV infection compared with the healthy controls. Furthermore, miRNA features including length distribution and end variations were characterized. Annotation of targets revealed a broad range of biological processes and signal transduction pathways regulated by HCV-induced fibrosis miRNAs. In addition, miRNAs of HCV-related fibrosis and control healthy were confirmed using miRNA microarray analysis. Real-time quantitative PCR (qPCR) analysis of miRNA in the chronic HCV infection patients and control healthy groups showed good concordance between the sequencing and qPCR data. This study provides the first large-scale identification and characterization of HCV-related fibrosis miRNAs and their potential targets, and represents a foundation for further characterization of their roles in the regulation of the diversity of HCV-related fibrosis.
