摘要
目的探讨富含亮氨酸重复序列免疫球蛋白样蛋白2(LRIG2)基因全长及胞外段对脑胶质瘤细胞系U251细胞增殖和凋亡的影响。方法将LRIG2全长,LRIG2胞外段(LRIG2ecto)及空白质粒(对照)经慢病毒法分别转染胶质瘤细胞系U251细胞,筛选稳定细胞株,实时PCR及免疫印迹法检测mRNA及蛋白表达,细胞增殖检测试剂盒检测细胞增殖,流式细胞分析检测细胞周期及细胞凋亡。结果两转染组Flag标签蛋白表达成功;两转染组LRIG2全长及LRIG2ectomRNA表达较对照组明显升高(P<0.01);两转染组细胞增殖率均明显高于对照组(P<0.01);两转染组S期与G2/M期细胞数之和(即细胞增殖指数)均明显高于对照组(P<0.01),而细胞凋亡率均明显低于对照组(P<0.05)。结论 LRIG2基因全长及LRIG2ecto促进胶质瘤细胞增殖,并将细胞周期阻滞于G2/M期,抑制细胞凋亡。
Objective To explore the effects of the leucine-rich repeats and immunoglobulin-like domains-2 （LRIG2） gene full length and LRIG2 ectodomain （LRIG2_ecto） on proliferation and apoptosis of glioma cell line U251 cells. Methods The lentiviral vectors expressing the full length andLRIG2_ecto were transfected into glioblastoma cell line U251 cells respectively. The expression levels of LR1G2 and LRIG2_ecto mRNA and protein in the U251 cells were determined respectively by realtime PCR and western blotting. The U251 cells proliferation was determined by cell counting kit-8 （CCK-8）. The U25l cells cycle distribution and apoptosis were detected by flow cytometry. Results The Flag proteins were successfully expressed in both the transfected groups. The mRNA expressions levels of LRIG2 andLRIG2_ecto, were significantly higher in the transfected U251 cells than those in the control ceils （P〈0.01）. The proliferation rates were significantly higher in the transfected U251 cells than those in the control cells （P〈0.01）. The proliferation indexes were significantly higher and the apoptotic rates were significantly lower in the transfected U251 ceils than those in the control cells （P〈0.01）. ConclusiQn Both LRIG2 full length andLRIG2_ecto may promote the proliferation and inhibit the apoptosis of glioma cell line U251 cells.
出处
《中国临床神经外科杂志》
2013年第1期30-33,共4页
Chinese Journal of Clinical Neurosurgery
基金
国家自然科学基金资助项目(NO:81001116)
