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目的探讨HCV核心蛋白对Hep G2细胞microRNA表达谱的影响。方法选择我国流行的HCV RNA(1b型),通过RT-PCR方法扩增出HCV-1b-C基因,双酶切后将其连接到真核表达载体pc DNA3.1(-)上,构建真核表达质粒pc DNA3.1(-)/HCV-1b-C;然后利用脂质体将上述真核表达载体转染至Hep G2细胞,并分为3组:空白对照组、pc DNA3.1(-)组及pc DNA3.1(-)/HCV-1b-C组,对其microRNA的表达及相关蛋白进行检测、分析,并探讨HCV核心蛋白对Hep G2细胞microRNA表达谱的影响。结果 pc DNA3.1(-)/HCV-1b-C真核表达载体得以成功构建,且对Hep G2细胞进行转染之后,HCV-C microRNA及蛋白得以成功转录及表达;pc DNA3.1(-)/HCV-1b-C组的microRNA及其表达程度相对来说较小,分别与其他2组相比,差异均具有统计学意义(P<0.05),但另2组差异均无统计学意义(P>0.05)。结论 HCV核心蛋白会影响Hep G2细胞microRNA表达,推测这可能与HCV感染人体后抑制细胞凋亡的机制存在一定的关系。
Objective To investigate the effect of HCV core protein on the expression of microRNA in Hep G2 cells. Methods HCV-1b-C gene was amplified by RT-PCR from HCV RNA in our country (type 1b). After double enzyme digestion, the HCV-1b-C gene was ligated into pcDNA3.1 (-) and the eukaryotic expression vector pcDNA3.1 The recombinant plasmid pcDNA3.1 (-) / HCV-1b-C was constructed and transfected into Hep G2 cells by lipofectamine. The recombinant plasmid was divided into 3 groups: blank control group, pcDNA3.1 -) group and pc DNA3.1 (-) / HCV-1b-C group. The microRNA expression and related proteins were detected and analyzed, and the effect of HCV core protein on the microRNA expression profile of Hep G2 cells was also investigated. Results The pcDNA3.1 (-) / HCV-1b-C eukaryotic expression vector was constructed successfully and HCV-C microRNA and protein were successfully transcribed and transfected into Hep G2 cells. The pcDNA3.1 (- ) / HCV-1b-C group, the difference was statistically significant (P <0.05), but the other two groups had no statistical significance (P> 0.05). Conclusion The HCV core protein may affect the microRNA expression in Hep G2 cells, suggesting that this may be related to the mechanism of HCV core inhibition of apoptosis.