论文部分内容阅读
目的:应用基因表达谱芯片(基因芯片)技术,检测乙型肝炎病毒DNA聚合酶RNaseH结构域蛋白(HBVDNAP RNaseH ,RNaseH)的表达对肝母细胞瘤细胞HepG2基因表达谱的影响,进一步阐明RNaseH对肝细胞基因表达的调节机制及其生物学功能。方法:以常规的分子生物学技术构建真核表达载体pcDNA 3 1(-) RNaseH ,以脂质体转染肝母细胞瘤细胞系HepG2 ,提取mRNA ,逆转录为cDNA ,与转染空白表达载体pcDNA 3 1(-)的HepG2细胞进行cDNA芯片分析。结果:RNaseH表达质粒转染的细胞有2 2 2条差异表达基因,其中113条基因表达水平上调,10 9条基因表达水平下调。结论:应用基因芯片成功筛选了RNaseH转染细胞后差异表达基因,为进一步阐明RNaseH的反式激活作用及免疫调节机制提供了新的依据。
OBJECTIVE: To detect the effect of HBVDNA RNaseH (RNaseH) on HepG2 gene expression in Hepatoblastoma cells by gene expression microarray (microarray), and to further elucidate the effect of RNaseH Regulatory Mechanism of Hepatocyte Gene Expression and Its Biological Function. METHODS: The eukaryotic expression vector pcDNA3 1 (-) RNaseH was constructed by conventional molecular biology techniques. The hepatoblastoma cell line HepG2 was transfected by lipofectamine 2000. The mRNA was reverse transcribed into cDNA and transfected into the blank vector pcDNA3 1 (-) HepG2 cells for cDNA microarray analysis. RESULTS: Two hundred and twenty-two differentially expressed genes were transfected by RNaseH expression plasmid, of which 113 genes were up-regulated and 109 genes were down-regulated. Conclusion: The gene differentially expressed in RNaseH transfected cells was successfully screened by gene chip, which provided a new basis for elucidating the transactivation and immunoregulatory mechanisms of RNaseH.