HCV NS4A与钙离子信号调节亲环素配体相互作用对细胞基因表达的影响

Detection of gene expression influence after the interaction between HCV NS4A and CAML with microarray assay

目的探讨HCV NS4A与钙离子信号调节亲环素配体(CAML)相互作用后对细胞基因表达的影响。方法分别构建HCV NS4A及CAML真核表达载体pCMV/Myc-NS4A及pcDNA 3.1/HisA-CAML,分别转染pCMV/Myc-NS4A(实验组)及pCMV/Myc空载体(对照组)至稳定表达CAML的293细胞,提取总mRNA,逆转录为cDNA,基因芯片技术检测转染NS4A对稳定表达CAML细胞株基因表达谱的影响。结果与细胞凋亡有关的Rho蛋白信号转导相关基因有7种,4种呈现上调表达,3种呈现下调表达。结论转染HCV NS4A后的293-CAML细胞的基因表达谱发生了一系列改变,尤其是一些与细胞凋亡相关的Rho蛋白信号转导途径相关基因。HCV NS4A可能通过与CAML相互作用对细胞凋亡过程产生影响,并且主要以负调节为主,从而赋予细胞对凋亡的抵抗,并可能因此导致病毒感染的慢性化。

Objective To study gene expression influence after the interaction between HCV NS4A and CAML.Methods The vector of pCMV/Myc-NS4A and pcDNA3.1/HisA-CAML were constructed by routine molecular biological methods.The 293 cells were transfected with pCMV/Myc-NS4A and pcDNA3.1/HisA-CAML by lipofectamine,respectively.And pCMV/Myc vector was transfected as negative control Total RNA was isolated and reverse transcribed.The cDNAs were subjected for microarray screening.Results Seven genes were found related to cell apoptosis,among which,4 showed up-regulated and 3 were down-regulated.Conclusions The expression of NS4A protein influenced the expression spectrum of 293-CAML cells,especially some genes related to cell apoptosis pathway.HCV NS4A may have some negative effect on the progress of cell apoptosis,which may lead to chronic HCV infection.