羟氯喹对人乳腺癌MDA-MB-435细胞增殖及凋亡的影响  

Influence of hydroxychloroquine on the proliferation and apoptosis of breast cancer cell MDA-MB-435

作　　者：张倩雯[1] 张媛媛[1] 晁振华[1] 张配[1] 蒋琛琛[1] 刘浩[1] 蒋志文[1] 

机构地区：[1]蚌埠医学院药学系,安徽省生化药物工程技术研究中心,安徽蚌埠233030

出　　处：《中国药理学通报》2013年第11期1549-1553,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81000992;81072207);安徽省高校省级自然科学研究项目(No KJ2012A202)

摘　　要：目的观察羟氯喹(hydroxychloroquine,HCQ)对人乳腺癌MDA-MB-435细胞增殖及凋亡的影响。方法采用MTT法及克隆形成抑制实验观察HCQ对人乳腺癌MDAMB-435细胞的增殖抑制作用;采用倒置显微镜观察HCQ处理后细胞形态学变化;采用Hoechst 33258荧光染色检测HCQ对人乳腺癌MDA-MB-435细胞凋亡的作用;采用PI单染流式细胞术检测细胞晚期凋亡水平;采用线粒体膜电位检测试剂盒(JC-1)观察HCQ处理人乳腺癌MDA-MB-435细胞后线粒体膜电位的转变。结果不同浓度的HCQ作用于乳腺癌MDA-MB-435细胞24、48、72 h后,均能明显抑制细胞增殖;在倒置显微镜下可观察到HCQ作用于乳腺癌MDAMB-435细胞后细胞逐渐萎缩变圆,细胞密度变小,失去正常细胞形态;HCQ能明显抑制MDA-MB-435细胞克隆形成;Hoechst 33258荧光染色结果显示,HCQ可以诱导乳腺癌MDA-MB-435细胞凋亡;PI结果显示,20、40、60μmol·L-1HCQ处理细胞24 h后,晚期凋亡率分别为7.1%、15.9%、35.3%(P<0.05);HCQ作用于乳腺癌MDA-MB-435后JC-1从红色荧光到绿色荧光转变(P<0.05)。结论 HCQ可以明显抑制人乳腺癌MDA-MB-435细胞增殖及诱导凋亡。Aim To observe the influence of hydroxychloroquine （HCQ） on the proliferation and apoptosis of breast cancer cell MDA-MB435. Methods The effect of HCQ on the proliferation of MDA-MB-435 cell was evaluated by M3T assay and clone formation assay; morphological changes of MDA-MB435 cell after HCQ treatment was observed by inverted microscope; Hoechst 33258 staining was used to assess the changes in HCQ-induced apoptosis of MDA-MB-435 cell; PI staining was used to analyze the apoptosis of the cell exposed to HCQ; JC-1 was used to detect the changes of mitochondrial membrane potential of MDA-MB435 cell. Results Incubation with HCQ for 24 h,48 h and 72 h resulted in a significant inhibition of MDA-MB- 435 cell proliferation; HCQ treatment at different concentrations caused MDA-MB-435 cell turning round and shrinkage gradually, density of cell turning smaller gradually and losing the normal cell morphology; HCQ treatment also inhibited cell colony formation; Hoechst 33258 staining showed HCQ could induce apoptosis of MDA-MB-d35 cell; exposure to 20,40,60 μmol · L^-1 HCQ for 24 h caused a percentage of late apoptotic cells of 7.1%,15.9%,35.3% （P 〈0.05）; HCQ treatment at different concentrations also turned JC-1 from red to green（P 〈0.05）. Conclusion HCQ can significantly inhibit the proliferation of MDA-MB-435 cell and induce apoptosis.

关 键 词：羟氯喹 增殖 凋亡 MDA-MB435细胞 流式细胞术 JC-1 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]