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Effects of hepatitis C virus genotype 1b core protein on the gene expressions of Bax and Bcl-2 in HepG2 cells
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摘要: 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...Abstract: 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 protein was detected by RT-PCR and Western Blot respectively.And then, detect the effects of HCV-1b-C on the expression of Bax and Bcl-2 gene in the HepG2 cell by RT-PCR and Western Blot, and set groups transfect with the plasmid pcDNA3.1 (-) and normal groups as control.Results The expression vector pcDNA3.1 (-) / HCV-1b-C was constructed successfully;And then, HCV C protein was expressed in the HepG2 cell line by transfection;For the corresponding mRNA and protein expression of Bax were lower in the groups transfected with HCV-1b-C compare to those in the groups transfected with the empty plasmid and normal groups (P<0.01) ;For the corresponding mRNA and protein expression of Bcl-2 were higher in the groups transfected with HCV-1b-C than those of in groups transfected with the empty plasmid and normal groups (P<0.01) .Conclusion The expression of genotype 1b HCV C in the HepG2 cell can reduce the expression of Bax and increase the expression of Bcl-2, leading to decrease in the ratio of Bax /Bcl-2, considering one of the mechanism of HepG2 cell escape from cell apoptosis.
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Key words:
- hepacivirus /
- viral core protein /
- hepatocytes /
- bcl-2 associated X protein /
- genes /
- bcl-2
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