应用微阵列技术筛选PS1TP1基因转染细胞差异表达基因

阐明乙型肝炎病毒(HBV)前S1蛋白反式激活蛋白1(PS1TP1)的表达对于肝细胞的基因表达谱的影响。应用基因芯片技术对于pcDNA3.1()和pcDNA3.1()PS1TP1分别转染的HepG2细胞的基因表达谱进行分析。以肝癌细胞系HepG2基因作为模板,应用聚合酶链反应(PCR)技术扩增PS1TP1基因片段,以常规的分子生物学技术构建表达载体pcDNA3.1()PS1TP1。以脂质体技术转染肝母细胞瘤细胞系HepG2,提取总RNA,逆转录为cDNA,与转染空白表达载体pcDNA3.1()的HepG2细胞进行DNA芯片分析并比较。在4096个基因表达谱的筛选中,发现有8个基因表达水平显著上调,14个基因表达水平显著下调。PS1TP1基因的表达对于肝细胞基因表达谱有显著影响。DNA芯片技术是分析反式调节靶基因的有效技术途径。

Screening of Genes Differentially Expressed in HepG2 Cells Transfected with Gene 1 Transactivated by Hepatitis B Virus Pre-s1 with Microarray Assay

To understand the differentially expressed target genes in HepG2 cells transfected with PS1TP1 protein expression vector, we compared the differentially expressed genes between the hepatoblastoma cell line HepG2 transfected by pcDNA3.1(-) and pcDNA3.1(-)-PS1TP1, respecti vely with cDNA microarray . The PS1TP1 coding DNA fragment was amplified with polymerase chain reaction (PCR) . The expressive vector of pcDNA3.1(-)-PS1TP1 was constructed by routine molecular biological methods. The HepG2 cells were transfected by pcDNA3.1(-) and pcDNA 3.1(-)-PS1TP1, respectively ,using FuGENE6 Transfection Reagent. Total RNA was isolated and reverse transcribed. The cDNAs were subjected for microarray screening with 4096 cDNA probes. From the scanning results, it was found that 8 genes were up-regulated and 14 genes were down-regulated by PS1TP1 protein expression. The expression of PS1TP1 protein affected the expression spectrum of hepatocyte.