摘要
目的:对新基因NS5A-TP4转染肝癌细胞的基因表达谱进行分析,探索该基因表达对肝细胞基因表达的调节机制及其生物学功能.方法:应用生物信息学(bioinformatics)技术,分析我室通过抑制性消减杂交技术筛选得到的新基因NS5A-TP4的全长编码序列,并NS5A-TP4构建基因的真核表达载体pcDNA3.1(-)-NS5A-YP4.应用基因表达谱芯片技术对重组表达质粒pcDNA3.1(-)-NS5A-TP4转染的HepG2细胞和空载体处理的相同细胞差异表达的mRNA进行检测.结果:确定基因NS5A-TP4由762nt组成,编码253aa的蛋白.基因表达谱芯片所检测的1152条目的基因均为GenBank中登录的基因,NS5A-TP4表达质粒转染的细胞有18条差异表达基因,其中12条基因表达增强,6条基因表达降低.这些差异表达的基因与细胞信号转导、凋亡、生长调节密切相关.结论:基因表达谱芯片技术可为初步探索新基因的功能提供重要的资料.本实验结果为进一步阐明NS5A-TP4生物学功能及HCV-NS5A的致病机制提供了理论依据.
AIM: To study the molecular mechanism of NS5A-TP4 in the up- and down-regulated genes of hepatocyte by cDNA microarray assay. METHODS: HepG2 cells were transfected by recombined expression plasmid pcDNA3.1(-)-NS5A-TP4. Total RNA was isolated from the transfected HepG2 cells with pcDNA3.1 (-) and pcDNA3.1(-)-NS5A-TP4, respectively. cDNA was prepared by reverse transcription. cDNA microarray was conducted for screening of up- and down-regulated genes of both HepG2 cells. RESULTS: After screening with cDNA microarray, we found that 12 genes were up-regulated, and 6 genes down-regulated. Some of them were involved in cell signal transduction, cell proliferation, and carcinogenesis. CONCLUSION: cDNA microarray is an important choice for the analysis of target genes of NS5ATP4 transactivation. The obtained sequences bring some new dues for studying the biological functions of NS5ATP4 and infection mechanism of HCV.
出处
《世界华人消化杂志》
CAS
2004年第2期311-314,共4页
World Chinese Journal of Digestology
基金
国家自然科学基金攻关项目
No.C03011402
No.C30070689军队"九
五"科技攻关项目
No.98D063军队回国留学人员启动基金项目
No.98H038军队"十
五"科技攻关青年基金项目
No.01Q138军队"十
五"科技攻关面上项目
No.01MB135~~
