摘要
目的研究过表达富含亮氨酸重复序列免疫球蛋白样蛋白1(leucine-rich repeats and immunoglobulin-like do-mains-1,LRIG1)对顺铂(cisplatin,CDDP)诱导的胶质瘤细胞U251凋亡的影响及其机制。方法应用脂质体介导的基因转染技术将对照质粒(EGFP-N1)及LRIG1质粒(EGFP-N1-LRIG1)分别转染胶质瘤细胞系U251,设为对照组和LRIG1过表达组,用Real-time PCR检测转染后LRIG1表达水平;用AnnexinⅤ/7AAD双标流式细胞术检测细胞凋亡;用Western blot检测各目的蛋白表达水平。结果 LRIG1过表达组LRIG1mRNA及蛋白均较对照组明显增高;0、10、20μg/mL CDDP作用下,LRIG1过表达组细胞凋亡率均明显高于对照组(P<0.01);随着CDDP浓度增加,胶质瘤细胞U251磷酸化表皮生长因子受体(P-EGFR)表达增加;不同浓度CDDP作用下,LRIG1过表达组P-EGFR水平均较对照组明显降低,促凋亡蛋白Bax,Caspase-3表达增加,抗凋亡蛋白Bcl-2表达降低。结论 LRIG1通过下调P-EGFR水平而促进顺铂诱导的胶质瘤细胞凋亡,增强胶质瘤细胞对顺铂的敏感性。
Objective To explore the effects of LRIG1 over-expression on the apoptosis of glioblastoma cells induced by cisplatin and the underlying mechanism.Methods The LRIG1 over-expression plasmid was transfected into glioblastoma cell line U251.The expression levels were detected by using Western blot and quantitative PCR.R-PE/7-AAD double labeling flow cytometry was used to detect the apoptosis rate.Western blot was used to investigate the expression levels of proteins.Results The mRNA and protein expression levels of LRIG1 were significantly increased in LRIG1 over-expression group as compared with control group.As compared with control group,the apoptosis rate in LRIG1 over-expression group was significantly increased.CDDP induced the activation of EGFR in a dose-dependent manner.The expression levels of P-EGFR were markedly decreased in LRIG1 over-expression group,the Bcl-2 protein was decreased,and the proapoptotic proteins,Bax and Caspase-3,were increased.Conclusion LRIG1 promotes the apoptosis of glioblastoma cells and enhances the cisplatin sensitivity by blocking CDDP-induced EGFR activation.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2012年第6期656-659,664,共5页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.81001116)
