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Tmub1过表达及沉默慢病毒载体转染对大鼠BRL-3A肝细胞增殖的影响
作者:赵晓彪 李光耀 刘孟刚 范霞 陈平 
单位:400042  重庆 第三军医大学大坪医院野战外科研究所肝胆外科(赵晓彪、李光耀、刘孟刚、陈平) 一室(范霞) 
关键词:基因 Tmub1 肝细胞 细胞增殖 慢病毒感染 
分类号:Q253
出版年,卷(期):页码:2016,41(1):12-16
摘要:

[摘要] 目的 探讨Tmub1蛋白在肝细胞增殖中的作用。方法 分别使用Tmub1沉默慢病毒载体、Tmub1过表达慢病毒载体及对应的空载体转染大鼠BRL-3A肝细胞,得到稳定转染的细胞。转染后采用Western blotting检测Tmub1蛋白的表达,MTT法检测细胞增殖情况,流式细胞术检测细胞周期。结果 转染LV-Tmub1慢病毒的肝细胞Tmub1蛋白表达明显增加,转染LV-Tmub1-RNAi慢病毒的肝细胞Tmub1蛋白表达明显降低,与正常肝细胞比较,差异均有统计学意义(P<0.01)。转染LV-Tmub1慢病毒的肝细胞增殖最快,转染LV-Tmub1-RNAi慢病毒的肝细胞增殖最慢,两组之间差异有统计学意义(P<0.01)。过表达Tmub1慢病毒载体转染的肝细胞的G2+S期明显长于沉默Tmub1慢病毒载体转染的肝细胞,差异有统计学意义(P<0.01)。结论 Tmub1蛋白可促进BRL-3A肝细胞的增殖。

[Abstract]  Objective  To investigate the role of Tmub1 protein in liver cell proliferation. Methods  Using silenced lentivirus (LV)-Tmub1 vector, over-expression LV-Tmub1 vector, and corresponding empty vectors to transfect rat's liver cell line BRL-3A to obtain stable transfected cells, which bear the expression of Tmub1 protein as detected by Western blotting, cell proliferation with MTT, and cell cycle with flow cytometry. Results  The Tumb1 protein expression in the hepatocytes transfected by LV-Tmub1 was increased significantly and that by LV-Tmub1-RNAi decreased significantly, with a statistically significant difference compared with normal hepatocytes (P<0.01). The proliferation was fastest in hepatocytes transfected by LV-Tmub1 and slowest in those by LV-Tmub1-RNAi, and the difference was statistically significant (P<0.01). The G2+S phase was longer in hepatocytes transfected by over-expression LV-Tmub1 vectors than in those by silenced LV-Tmub1 vectors, and the difference was statistically significant (P<0.01). Conclusion  Tmub1 protein can promote the hepatocyte proliferation in BRL-3A cell line.

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