细胞生物学综述综述标题.docVIP

乙型肝炎病毒X蛋白诱导肝卵圆细胞恶性转化 廖 凯，王月飞 ，卢乐乐， 张雷达(400038，重庆，第三军医大学西南医院肝胆外科) [摘要] 目的 研究乙型肝炎病毒X蛋白（Hepatitis B virus X, HBx）对肝卵圆细胞的转化作用。方法 构建表达HBx和嘌呤霉素抗性的慢病毒LV-HBx-PM及对照慢病毒LV-PM分别感染WB-F344肝卵圆细胞。以感染HBx慢病毒载体WB-F344细胞作为实验组（WB-F344-HBx），以感染对照慢病毒载体的WB-F344细胞作为阴性对照组（WB-F344-PM），以未转染慢病毒载体的培养相同代数的WB-F344细胞作为空白对照组（WB-F344-Mock）。通过real-time RT-PCR和Western blot验证各组细胞中HBx的表达；运用MTT法、流式细胞术、细胞迁移实验，裸鼠移植瘤实验观察各组细胞的增殖能力、细胞周期、迁移能力和成瘤能力。结果 WB-F344-HBx细胞在基因水平和蛋白水平稳定高效表达HBx；WB-F344-PM和WB-F344-Mock不表达HBx。与阴性对照组WB-F344-PM相比，实验组WB-F344-HBx G0/G1期细胞比例降低（55.98%±0.39%, 61.65%±1.37%, P=0.003），G2/M期细胞比例升高（15.05%±1.67%， 7.83%±0.46%， P0.001），细胞周期进程加快，细胞增殖能力增强（P0.001），具有更强的细胞迁移能力（P0.001）和成瘤能力；而阴性对照组WB-F344-PM与空白对照组WB-F344-Mock相比，细胞周期、细胞增殖能力、细胞迁移能力和成瘤能力无显著变化。结论 乙型肝炎病毒X蛋白可诱导肝卵圆细胞恶性转化。 [关键词] 乙型肝炎病毒；X蛋白；卵圆细胞；恶性转化 [中图分类号] R735.7 [文献标志码] A Hepatitis B virus X protein induces hepatic oval cell malignant transformation Liao Kai, Wang Yuefei, Lu Lele, Zhang Leida（Department of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, Chongqing 400038, China） [Abstract] Objective The aim of this study was to detect the malignant transformation of hepatitis B virus X (HBx) protein on hepatic oval cells. Methods The WB-F344 cell infecting with lentivirus expressing HBx and puromycin(PM)-resistant gene was de?ned as experimental group (WB-F344-HBx). The WB-F344 cell infecting with lentivirus only expressing puromycin-resistant gene was de?ned as negative control group (WB-F344-PM). The WB-F344 cell without infecting with lentivirus was de?ned as blank control group (WB-F344-Mock). The expressions of HBx in the three groups were confirmed by real-time RT-PCR and Western blot analysis. The cell cycle, proliferation, ability of metastasis and tumorigenicity were detected with flow cytometry, MTT assay, metastasis experiment and nude mouse transplantation tumor experiment, respectively. Results WB-F344-HBx highly stable expressed HBx protein while WB-F344-PM and WB-F344-Mock wi