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目的探讨丙型肝炎病毒非结构蛋白NS4B对肝细胞内p53表达的影响,以及在肝癌发生中的作用与机制。方法设置空白对照组、空白载体组、转染NS4B组、转染p53组、共转染NS4B及p53组。使用脂质体介导转染法,转染丙型肝炎病毒非结构蛋白重组质粒PCXN2-NS4B及突变型p53基因重组质粒pC53-CX22AN3进入Chang肝细胞内,并用G418筛选获得稳定表达细胞。采用免疫细胞化学法检测p53表达率。结果空白对照组无p53表达,空白载体组及转染NS4B组呈弱阳性表达,转染p53组及共转染组呈阳性表达;转染p53组、共转染组分别与空白对照组、空白载体组及转染NS4B组比较,差异均有统计学意义(P<0.05)。结论 NS4B可能抑制p53表达,也可能阻止其进入细胞核,但NS4B与突变型p53关系不明确。NS4B导致肝细胞异常增生,诱导肝癌发生可能不依赖p53的异常表达及突变。
Objective To investigate the effect of hepatitis C virus nonstructural protein NS4B on the expression of p53 in hepatocytes and its role in the pathogenesis of hepatocellular carcinoma. Methods The blank control group, blank vector group, transfected NS4B group, transfected with p53 group, transfected NS4B and p53 group. The recombinant plasmid pcXN2-NS4B and the mutant p53 gene plasmid pC53-CX22AN3 were transfected into Chang hepatocytes by liposome-mediated transfection, and stable expression cells were screened by G418. The expression of p53 was detected by immunocytochemistry. Results There were no p53 expression in blank control group, weak expression in blank vector group and NS4B transfected group, positive expression in transfected p53 group and co-transfected group, transfected with p53 group, transfected with blank control group, blank control group Compared with NS4B group, the difference was statistically significant (P <0.05). Conclusion NS4B may inhibit the expression of p53, may also prevent its entry into the nucleus, but the relationship between NS4B and mutant p53 is not clear. NS4B lead to abnormal proliferation of hepatocytes, liver cancer induction may not depend on the abnormal expression of p53 and mutations.