氯喹对食管癌细胞系放射增敏的实验研究  

Study of radiosensitization of chloroquine on esophageal cancer cell line

作　　者：袁晓丽[1] 李涛[1] 黄建鸣[2] 查晓[2] 邓碧芳[2] 郎锦义[1] 

机构地区：[1]四川省肿瘤医院食管腹部放疗病区,成都610041 [2]四川省肿瘤研究所分子生物实验室

出　　处：《中华放射医学与防护杂志》2014年第11期823-826,共4页Chinese Journal of Radiological Medicine and Protection

基　　金：四川省科技厅科技支撑项目

摘　　要：目的 探讨氯喹对食管癌TE-1细胞系的放射增敏作用及其主要机制，方法 采用MTT法检测不同浓度氯喹对TE-1细胞的生长抑制作用，分别用单纯照射或照射前联合氯喹、照射后联合氯喹作用于TE-1细胞,作用6h后用Western blot测定自噬相关蛋白LC3和Beclin1的表达,用Lyso-Tracker Red DND-99/Hochest 33258进行荧光染色,并用荧光显微镜观察细胞内酸性囊泡（AVOs）的变化，克隆形成实验分析细胞增殖的改变,拟合剂量-生存曲线并计算放射增敏参数，结果 氯喹对TE-1细胞的生长抑制作用呈浓度依赖性，辐射显著诱导TE-1细胞LC3和Beclin1的表达,并促进LC3-Ⅰ转换为LC3-Ⅱ，与单纯照射及照射前加药相比,照射后加药显著降低TE-1细胞内AVOs的荧光强度（F=16.44,P＜0.05）和克隆形成率,照射前加药和照射后加药的SERD0分别为1.037和1.439（t =8.30,P＜0.05），结论 氯喹能增加食管鳞癌TE-1细胞的放射敏感性,其机制可能与抑制自噬作用相关。Objective To investigate the possibility of chloroquine radiosensitization of esophageal cancer cell line TE-1 and its further mechanism.Methods Effect of chloroquine on cell viability of TE-1 cells was determined by MTT method.Expression of LC3,Beclin-1 and formation of acidic vesicular organelles （AVOs） were determined by Western blot,and fluorescence staining with Lyso-Tracker Red DND-99,respectively.Clonogenic survival of TE-1 cells was examined by clonogenic forming assay.Results Chloroquine showed dose-dependent inhibition of TE-1 cell growth,and its values of IC50 and IC10 were （72.33 ± 5.28） and （15.42 ± 3.33） μmol/L,respectively.The expression of Beclin-1 and LC3-Ⅱ/Ⅰ markedly increased in irradiated TE-1 cells.The addition of chloroquine with IC10concentration significantly reduced the fluorescence and intensity of AVOs accumulation in the cytoplasm of TE-1 cells.Clonogenic survival fraction decreased obviously,in TE-1 cells with addition of chloroquine after radiation and the value of SERD0 was 1.439.Conclusions Chloroquine could radiosensitize esophageal cancer cells by blocking autophagy-lysosomal pathway and be used as a potential radiosensitizing strategy.

关 键 词：食管肿瘤 自噬 氯喹 放射敏感性 

分 类 号：R735.1[医药卫生—肿瘤]