氯喹对正常胃上皮细胞GES-1和胃癌细胞HGC-27凋亡的不同影响  

Inconsistent effect of chloroquine on apoptosis of normal gastric epithelial cells GES-1 and gastric cancer cell line HGC-27

作　　者：曹丽 苏强[2] 张亚萍 张鑫艳 程凯[3] 张晓延[1] CA O Li;SU Qiang;ZHANG Ya-Ping;ZHANG Xin-Yan;CHENG Kai;ZHANG Xiao-Yan(Fenyang College of Shanxi Medical University, Fenyang 032200, Chin)

机构地区：[1]山西医科大学研究生院,太原030001 [2]山西医科大学第一医院神经内科,太原030001 [3]山西医科大学汾阳学院,汾阳032200

出　　处：《中国免疫学杂志》2018年第6期810-814,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金(No.81301426); 山西医科大学汾阳学院校内科研项目(No.2018C01); 山西省高等学校大学生创新创业训练项目(No.2014499)资助

摘　　要：目的:比较氯喹对正常胃上皮细胞GES-1和胃癌细胞HGC-27凋亡的不同影响。方法:使用倒置显微镜观察CQ处理后这两种细胞形态学变化;采用DAPI核染色检测CQ对HGC-27细胞凋亡的作用;运用氯喹和雷帕霉素作用这两种细胞72 h后,用CCK-8检测这两种药物对两种细胞的增殖活性;JC-1检测氯喹处理后细胞线粒体膜电位的变化;用免疫印迹法检测凋亡效应酶Caspase-3和底物PARP的改变。结果:10μmol/L的CQ作用于GES-1细胞和HGC-27细胞72 h后,在镜下可见,氯喹对GES-1细胞的形态无明显影响,却能使HGC-27细胞间隙增宽,悬浮细胞数目逐渐增多,细胞密度明显减少,细胞逐渐萎缩变圆,胞质减少,失去正常细胞形态;通过DAPI核染色发现,氯喹作用两种细胞72 h后,GES-1细胞核浅染、核大小形态均未发生变化,HGC-27细胞核呈浓缩致密的固缩形态或颗粒状荧光;CQ和RAP作用于正常胃上皮细胞和胃癌细胞HGC-27 72 h后,CCK-8结果显示相对于正常胃上皮GES-1细胞而言,氯喹能抑制胃癌HGC-27细胞的增殖活性;JC-1结果显示氯喹作用于HGC-27细胞后发生了红色荧光向绿色荧光的转变;Western blot显示胃癌细胞HGC-27的凋亡蛋白Caspase-3和PARP表达显著减少。结论:相比正常胃上皮细胞GES-1,CQ可以明显抑制人胃癌细胞HGC-27细胞活力并诱导凋亡。Objective：To compare the effect of chloroquine on apoptosis of normal gastric epithelial cells and gastric cancer cell line HGC-27.Methods：Change of these two kinds of cells were observed by inverted microscope after treating with CQ.HGC-27 cells were detected on the effect of apoptosis by DAPI nuclear staining after treating with CQ.The proliferation of cells were measured by CCK-8.Changes of mitochondrial membrane potential were investigated by JC-1 after treating with CQ.The expression of apoptosis protein effector enzyme Caspase-3 and substrate PARP in these two kinds of cells were tested by Western blot after using chloroquine（CQ） and rapamycin（rapamycin,RAP） to treat cells 72 h.Results：After treated with 10 μmol/L CQ 72 h,morphological characteristics of GES-1 cells and HGC-27 cells could be visible under the microscope,CQ induced apoptosis of GES-1 cells,on the contrary,it could make the HGC-27 cell get widened,the number of apoptotic cells gradually increased,the cell density decreased,cell atrophy and gradually turned round,cytoplasm reduced,at last,lose normal cell morphology.After two kinds of cells treated with CQ 72 h,as for GES-1 cell nuclei stained light,nuclear size and shape were not changed,however,HGC-27 nuclei showed pyknosis or granular fluorescence dense concentrated form.CCK-8 results showed that comparing with normal gastric epithelial cells GES-1,the proliferation of gastric cancer HGC-27 cells activity could be inhibited by CQ.JC-1 results showed that the change of the red fluorescence to green fluorescence in HGC-27 cells treated by CQ.Western blot showed that after being treated with CQ and RAP in normal gastric epithelial cells and HGC-27 cell line 72 h,the expression of apoptosis protein Caspase-3 and PARP in gastric cancer cell HGC-27 decreased significantly,comparing to that in GES-1 cells.Conclusion：Compared to normal gastric epithelial cells,CQ can inhibit human gastric cancer HGC-27 cell viability and induce apoptosis.

关 键 词：氯喹 凋亡 GES-1细胞 HGC-27细胞 

分 类 号：R392.12[医药卫生—免疫学] R735.2[医药卫生—基础医学]