HCV核心基因转染对QBC939细胞凋亡及细胞周期的影响

The Effect of Hepatitis C Virus Core Protein on Apoptosis and Cell Cycle of Hilar Cholangiocarcinoma Cells

【目的】研究丙型肝炎病毒核心蛋白(HCV-C)对肝门部胆管癌细胞(QBC939)细胞凋亡及细胞周期的影响。【方法】将包含HCV-C基因的真核表达质粒pcDNAHCV-C和空载体,利用脂质体介导将其转染QBC939,经G418筛选获得稳定转染HCV-C的QBC939细胞(QBC939HCV-C+),RT-PCR和免疫荧光法检测HCV-C蛋白表达,MTT法检测QBC939HCV-C+细胞、空白质粒转染QBC939(QBC939pcDNA)细胞和未转染QBC939细胞生长增生率;流式细胞术(FACS)检测3组细胞凋亡率和细胞周期,以及QBC939HCV-C+细胞经Fas抗体诱导凋亡后的细胞周期变化。【结果】①QBC939HCV-C+细胞增殖率显著高于空白质粒转染QBC939和未转染QBC939细胞增殖率;②细胞未经Fas抗体诱导凋亡时,QBC939HCV-C+细胞S期(20.1%±1.8%)高于未转染QBC939细胞S期(14.2%±3.6%),QBC939HCV-C+细胞凋亡率(5.0%±0.7%)低于无HCV-C转染QBC939细胞凋亡率(9.2%±0.7%);③QBC939HCV-C+细胞经Fas抗体诱导凋亡时,细胞S期低于未经诱导凋亡细胞S期。【结论】HCV-C蛋白具有抑制QBC939细胞凋亡和促进细胞增殖作用。

To explore the effect of hepatitis C virus core protein (HCV C) on apoptosis and cell cycle of hilar cholangiocarcinoma cells(QBC939). The recombinant plasmid (pcDNA HCVC) including the gene of HCV C and the vector alone were transfected into QBC939 with liposome. Then it was selected with G418 and resistant colonies (QBC939 HCV C+) were obtained. The colonies were identificated by reverse transcription PCR (RT PCR) and immuohistochemistry. The growth rate of QBC939 HCV C+, QBC939 pcDNA and QBC939 were detected by MTT assay. The apoptosis rate and cell cycle of this three group cells were detected by flow cytometry (FACS). The change of cell cycle of QBC939 HCV C+which was induced apoptosis by Fas antibody was also detected by FACS.①The cell proliferation rate of QBC939 HCV C+was significantly higher than that of QBC939 pcDNA and QBC939; ②When all cells were not induced by Fas antibody, the S stage percentage(20.1%±1.8%)of QBC939 HCV C+was higher than that of QBC939(14.2%±3.6%)and the apoptosis rate of QBC939 HCV C+was lower than that QBC939 without being tansfected by HCV C. ③When the QBC939 HCV C+was induced apoptosis by Fas antibody, the apoptosis rate of QBC939 HCV C+was lower then that of the cells without being induced [Conclusion]HCV C protein may contribute to hilar cholangiocarcinoma cell proliferation and reduce hilar cholangiocarcinoma cells apoptosis.