CAJ | 학술논문

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.
목적 관찰병형간염병독핵심단백(HCVc)재유도담관암세포상피-간협양표형전화중적작용.방법 장전염적담관암세포(QBC939)분성공재체대조조화HCVc기인실험조,통과반정량역전록-취합매련반응(RT-PCR)화면역세포화학적방법 분별검측량조세포중HCVc、상피성표지물상피성개점부소(E-cadherin)、간협성표지물파형단백(Vimentin)、섬유련접단백(Fibronectin)적mRNA급단백적표체.결과 전염HCVc기인적실험조세포능구교호지표체해기인,이공재체대조조미견기표체,실험조세포E-cadherin적mRNA표체수평(0.44±0.03)교대조조(0.56±0.03)명현하강(P＜0.05),이기Vimentin(0.61±0.01)、Fibronectin(0.58±0.05)표체수평교대조조(0.38±0.02)、(0.42±0.03)명현증가(P＜0.05).량조세포각표지물단백표체차이명현,전염HCVc기인적실험조세포교공재체대조조세포E-cadherin표체명현결실,Vimentin、Fibronectin명현증고.결론 병형간염병독핵심단백가능유도담관암세포발생료상피-간협양표형전화.
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.