氯喹通过抑制自噬促进TRAIL诱导的胶质瘤细胞凋亡  

Chloroquine promotes TRAIL-induced apoptosis of glioma cells through inhibition of autophagy

作　　者：冯驰[1] 郭双毅 程龙海[2] 罗杰[2] 

机构地区：[1]武汉大学中南医院神经外科,武汉420073 [2]十堰市太和医院神经外科,湖北442000

出　　处：《中国临床神经外科杂志》2016年第4期227-229,共3页Chinese Journal of Clinical Neurosurgery

摘　　要：目的探讨氯喹对肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导的胶质瘤细胞凋亡的影响。方法体外培养人脑胶质瘤细胞U251和Hela细胞,分为对照组、氯喹组、重组人TRAIL蛋白(rhTRAIL)组和联用组(氯喹与rhTRAIL联合应用);MTT法检测细胞活力,Annexin V-FITC/PI凋亡检测试剂盒检测细胞凋亡,共聚焦显微镜检测GFP-LC3的分布判断细胞自噬水平,Western Blot检测cleaved caspase-8的表达水平。结果氯喹和rhTRAIL均显著增加U251细胞和Hela细胞自噬水平,而且联用较单独应用自噬水平更高,Hela细胞自噬水平明显高于U251细胞。U251细胞和Hela细胞增殖抑制率和细胞凋亡率:氯喹组与对照组均无统计学差异(P>0.05),rhTRAIL和联用组均明显高于氯喹组(P<0.05),联用组明显高于rhTRAIL组(P<0.05)。氯喹组和对照组Cleaved caspases-8水平无统计学差异(P>0.05),rhTRAIL和联用组均明显增高(P<0.05),联用组明显高于rhTRAIL组(P<0.05)。结论氯喹能通过抑制细胞自噬,增强TRAIL诱导的U251细胞凋亡。Objective To study the mechanism of glioma cells apoptosis induced by Chloroquine（CQ）.MethodsHuman glioma U251 cells were treated with recombined human tumor necrosis factor-related apoptosis-inducing ligand（rh TRAIL） and/or CQ. The cells vitality was detected by MTT assay and the U251 cells apoptosis was detected by Annexin V-FITC/PI Apoptosis Detection Kit. The GFP-LC3 distribution was detected by confocal microscope. The expression level of caspases-8 was detected by Western Blot.Results The autophagy level of the U251 cells treated with rh TRAIL increased when rh TRAIL induced the U251 cells apoptosis. The growth inhibition rate [（47.11±0.18）%] and the cell apoptosis rate [（34.31±0.97）] of U251 cells treated with rh TRAIL and CQ were significantly higher respectively than those [（23.88±0.48）% and（19.68±0.67）% respectively] of U251 cells treated with rh TRAIL only（P〈0.05）. The level of cleared caspases-8 expression was significantly higher in the U251 cells treated by rh TRAIL and CQ than those in U251 cells treated by rh TRAIL only（P〈0.05）.ConclusionIt is suggested that CQ may enhance rh TRAIL-induced apoptosis through inhibiting the autophagy induced by rh TRAIL.

关 键 词：胶质瘤 U251细胞 肿瘤坏死因子相关凋亡诱导配体 氯喹 细胞凋亡 细胞自噬 

分 类 号：R739.41[医药卫生—肿瘤] R730.59[医药卫生—临床医学]