转染丙型肝炎病毒核心基因对胆管癌细胞上皮-间叶样表型转化的影响

目的 观察丙型肝炎病毒核心蛋白(HCVc)在诱导胆管癌细胞上皮-间叶样表型转化中的作用.方法 将转染的胆管癌细胞(QBC939)分成空载体对照组和HCVc基因实验组,通过半定量逆转录-聚合酶链反应(RT-PCR)和免疫细胞化学的方法 分别检测两组细胞中HCVc、上皮性标志物上皮性钙黏附素(E-cadherin)、间叶性标志物波形蛋白(Vimentin)、纤维连接蛋白(Fibronectin)的mRNA及蛋白的表达.结果 转染HCVc基因的实验组细胞能够较好地表达该基因,而空载体对照组未见其表达,实验组细胞E-cadherin的mRNA表达水平(0.44±0.03)较对照组(0.56±0.03)明显下降(P＜0.05),而其Vimentin(0.61±0.01)、Fibronectin(0.58±0.05)表达水平较对照组(0.38±0.02)、(0.42±0.03)明显增加(P＜0.05).两组细胞各标志物蛋白表达差异明显,转染HCVc基因的实验组细胞较空载体对照组细胞E-cadherin表达明显缺失,Vimentin、Fibronectin明显增高.结论 丙型肝炎病毒核心蛋白可能诱导胆管癌细胞发生了上皮-间叶样表型转化.

The effect of hepatitis C virus core gene transfection on epithelial-mesenchymal transition in human biliary carcinoma cell line

Objective To observe the effect of hepatitis C virus core protein (HCVe) on epithelial-mesenehymal transition (EMT) in human biliary carcinoma cell line. Methods After The PGST HCVcI95 plasmid containing HCVc full long gene (experimental group) and the PCEX-3ks (empty vector, eantrol group) were transfected into QBC939 cells respectively, the expression of HCVe, E-cadherin, Vimentin, Fibronectin mRNA and protein was detected by RT-PCR and immunocytochemistry, respectively. Results The high expression of HCVc mRNA and protein was observed in experimental group, but not in control group. The E-cadherin expression (0.44 ± 0.03 vs 0.56 ± 0.03 ) was decreased ( P ＜ 0.05 ), but the expression of Vimentin (0.61 ±0.01 vs 0.38 ± 0.02) and Fibronectin (0.58 ±0.05 vs 0.42 ± 0.03) was increased (P ＜ 0.05 ) in experimental group as compared with that in control group. Conclusion HCVe may contribute to epithelial mesenchymal transition in human biliary carcinoma cells.