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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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[1]Lee S, Park U, Lee YI.Hepatitis C virus core protein transactivates insulin-like growth factor II gene transcription through acting concurrently on Egr1 and Sp1 sites[J].Virology, 2001, 283 (2) :167-177. [2]Nguyen H, Sankaran S, Dandekar S.Hepatitis C virus core protein induces expression of genes regulating immune evasion and anti-apoptosis in hepatocytes[J].Virology, 2006, 354 (1) :58-68. [3]刘妍, 成军.HCV致肝细胞癌的分子生物学机制[J].国外医学·流行病学传染病学分册, 2000, 27 (1) :10-13. [4]Schwer B, Ren S, Pietschmann T, et al.Targeting of hepatitis C virus core protein to mitochondria through a novel C-terminal localization motif[J].Virol, 2004, 78 (15) :7958-7968. [5]朱德强, 黄志勇.DNA损伤与肝癌发生[J].世界华人消化杂志, 2007, 15 (16) :1775-1780. [6]Giannini C, Caini P, Giannelli F, et al.Hepatitis C virus core protein expression in human B-cell lines does not significantly modify main proliferative and apoptosis pathways[J].Gen Virol, 2002, 83 (Pt 7) :1665-1671. [7]刘英, 郭世杰, 孙景辉, 等.缺氧缺血性脑损伤新生大鼠心肌细胞凋亡及凋亡蛋白Bax、Bcl-2表达的变化[J].临床儿科杂志, 2009, 27 (12) :1172-1176. [8]Otsuka M, Kato N, Taniguchi H, et al.Hepatitis C virus core protein inhibits apoptosis via enhanced Bcl-xL expression[J].Virology, 2002, 296 (1) :84-93. -
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