丙型肝炎病毒1b基因型核心蛋白对HepG2细胞Bax与Bcl-2基因表达的影响

目的研究丙型肝炎病毒（HCV）1b基因型核心蛋白（C）对HepG2细胞B细胞淋巴瘤-2基因（Bcl-2）与Bcl-2相关X蛋白（Bax）表达的影响,以探索1b型HCV C蛋白与HepG2细胞凋亡的关系。方法利用RT-PCR扩增出HCV-1b-C基因,经双酶切后连接pcDNA3.1（-）,成功构建真核表达载体pcDNA3.1（-）/HCV-1b-C。利用脂质体转染HepG2细胞,RT-PCR及Western Blot检测其mRNA及蛋白的表达,RT-PCR及Western Blot检测转染成功后HCV-1b-C对HepG2细胞Bax与Bcl-2表达的影响,并设转染空质粒组及未处理组作对照。结果成功构建真核表达载体pcDNA3.1（-）/HCV-1b-C;瞬时转染HepG2细胞,成功表达HCV C mRNA及蛋白;转染C基因组的Bax的mRNA及蛋白相对表达量减少,与转染空质粒组及未处理组比较差异均有统计学意义（P〈0.01）;转染C基因组的Bcl-2的mRNA及蛋白相对表达量增多,与转染空质粒组及未处理组比较差异均有统计学意义（P〈0.01）。结论 1b基因型HCV C蛋白转染HepG2细胞会导致Bax表达减少及Bcl-2表达增多,降低Bax/Bcl-2比值,可能是抑制HepG2细胞凋亡的机制之一。

Effects of hepatitis C virus genotype 1b core protein on the gene expressions of Bax and Bcl-2 in HepG2 cells

Objective To investigate the effects of hepatitis C virus genotype 1b core protein on the expression of Bax and Bcl-2 in the HepG2 cell and to study the relationship of core protein and cell apoptosis.Methods The gene of HCV-1b-C was amplified by RT-PCR,and then it was inserted into pcDNA3.1(-) to construct the expression plasmid pcDNA3.1(-)/HCV-1b-C by double enzyme-cut.To transfect the HepG2 cell with recombination expression vector,pcDNA3.1(-)/ HCV-1b-C,by lipofectamine.The expression of HCV C mRNA and p...