乙型肝炎肝硬化基因表达谱的研究

目的利用寡核苷酸芯片测定乙型肝炎肝硬化基因表达谱,筛选肝硬化差异基因。方法应用含有18 993个已知基因的A ffym etrix U133p lus2.0芯片,对从肝硬化抽提及纯化标记的mRNA进行芯片杂交,用GeneP ix 4 000A荧光激光扫描系统对芯片进行扫描,GeneP ix 3.0分析软件处理杂交信号获取结果。以Northern印迹技术检验芯片分析结果的可靠性。结果芯片分析结果具有可重复性。肝硬化差异表达基因共有118个,上调基因63个,下调基因55个。结论寡核苷酸芯片是筛选肝硬化差异表达基因的有力工具,多基因差异表达参与肝硬化发生。

Profiles of gene expression in patients with cirrhosis due to chronic viral hepatitis B

Objective To study gene expression profiles in hepatitis B virus related liver cirrhosis by oligonucleotide microarray,and to identify gene differentially expressed in the cirrhosis.Methods Total RNA was extracted from cirrhosis and matched normal liver tissue which were histologically defined,and was reverse transcribed into double-strand cDNA.After cDNA was transcribed into biotinylated cRNA targets,purified and fragmented according to the manufacturer's instructions,the cRNA targets were hybridized to the oligonucleotide microarray containing 18 993 genes.The chips were scanned with GenePix 4000A,and the signals were analyzed with GenePix Pro 3.0.Northern blotting confirmed the results reliable.Results Using this methodology,118 genes,which expression level were more than 2 times were screened out.In the cirrhosis tissue,63 genes of them had higher expression and 55 of them had lower expression.Conclusion The oligonucleotide microarray is a powerful method to identify cirrhosis-associated genes,and many different kinds of genes are possibly involved in the initiation and progression of cirrhosis.