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

Effects of over-expression and silenced lentivirus Tmub1 vectors transfection on the proliferation of rat liver cell line BRL-3A

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。

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, w ith 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.