LRIG1基因表达下调对胶质瘤细胞周期及凋亡的调控作用

Effects of down-regulation of LRIG1 expression on glioma cells cycle and apoptosis

目的 探讨富含亮氨酸重复序列免疫球蛋白样蛋白1（LRIG1）基因表达下调后对胶质瘤细胞的周期及凋亡的影响及其机制.方法 将携带针对LRIG1基因特异性RNA干扰序列及非特异性shRNA编码序列的质粒载体分别转染GL15细胞株,用G418（600 mg/L）筛选出稳定株后,Western blot法检测LRIG1蛋白表达的改变,PI（0.05 g/L）与Rnasine（0.5 g/L）标记后流式细胞仪检测对照组与实验组细胞的周期差异,Annexin V-FTTC/PI双标后用流式细胞仪观察两组细胞凋亡的改变.结果 成功获得干扰LRIG1基因的GL15细胞稳定株,实验组较对照组LRIG1基因表达下降54.7%.流式细胞仪分析显示,实验组的G2/M期细胞百分率较对照组明显增高（P＜0.01）,实验组细胞的凋亡比例明显低于对照组（P＜0.01）.讨论抑制LRIG1表达后,GL15细胞的抗凋亡能力明显增强并能使细胞阻滞在G2/M期.

Objective To investigate the effects of down-regulation of LRIG1 expression on glioma cells cycle and apoptosis and the possible mechanism. Methods Small interfering RNA （siRNA） targeting LRIC1 gene and negative shRNA （neg） were constructed and transfected into CL1S glioma cells. And the cells （siRNA） that stably suppressed LRIG1 expression were selected by G418 （600 mg/L）. The change of LRIG1 protein level was measured by Western blotting. The apoptosis rate and cell cycle （0.05 g/L PI, 0.5 g/L RNasine） were analyzed by flow cytometry. Results The LRIG1 protein level in pGene-sil2-LRIGl -shRNA （siRNA） transfected cells was significantly silenced by 54.7%. The propotion of cells in G2/M phase was significantly higher, and apoptosis rate was obviously lower in experimental group than in control group （both P 〈 0.01）. Conclusion Down-regulation of LRIG1 expression can suppress the apoptosis of glioma cells and arrest the cells in G2/M phase.