抑制自噬增强MCF-7乳腺癌细胞对依托泊苷的敏感性  

Inhibition of autophagy increases the sensitivity of breast cancer MCF-7 cells to Etoposide

作　　者：刘晓健[1] 黄雷 陈睿琦[3] 张秀冰[3] 胡峰[3] 曾庆虹[3] 

机构地区：[1]辽宁医学院药理学教研室,辽宁锦州121001 [2]锦州市食品药品检验所,辽宁锦州121001 [3]辽宁医学院基础医学院,辽宁锦州121001

出　　处：《基础医学与临床》2016年第8期1087-1091,共5页Basic and Clinical Medicine

基　　金：辽宁省自然科学基金(2014022040);辽宁医学院校长基金-奥鸿博泽基金大学生科技创新项目(2014D04);地方高校国家级大学生创新创业训练计划(201510160009)

摘　　要：目的探讨自噬抑制剂(Baf)在乳腺癌MCF-7细胞对依托泊苷(VP-16)的药物敏感性的影响。方法实验分为对照组(NC)、Baf组、VP-16和VP-16+Baf组。用MTT法检测细胞生存活力、GFP-LC3质粒转染后荧光显微镜检测绿色荧光的分布、Western blot检测蛋白表达和流式细胞仪检测细胞凋亡。结果 15μmol/L的VP-16使细胞活力降低,而在收集细胞前12 h时加入10 nmol/L的Baf进一步降低了细胞活力(P<0.01);VP-16明显增加MCF-7细胞的LC3Ⅱ的表达和GFP-LC3绿色荧光斑点的聚集,而减少了P62的蛋白表达;与VP-16组比较,VP-16+Baf组细胞P62的蛋白表达增多,凋亡蛋白cleaved-PARP的表达和细胞凋亡比例也明显增多(P<0.01)。结论 VP-16抑制乳腺癌细胞增殖的过程中诱导了保护性自噬,抑制自噬促进了凋亡性死亡,可以增加癌细胞对VP-16的敏感性。Objective To investigate the effects of bafilomycin A1 （Baf） on the sensitivity of MCF-7 breast cancer cells to etoposide （VP-16）. Methods The cultured cells were divided into normal control （NC）, Bar, VP-16 and VP- 16 ＋ Baf groups. The viability of cells was determined with MTF assay. After GFP-LC3 plasmid transfection to the cells, the distribution of green fluorescence was observed by using fluorescence microscopy. The protein expres- sion was assayed by Western blot and the apoptosis of MCF-7 cells was detected by flow cytometry. Results VP-16 at 15 μmol/L reduced the viability of cells. Baf （ 10 nmol/L） which was added to treat the cells for 12 h before the cells collection reduced the viability of cells more than VP-16 used only（P 〈0. 01 ）. VP-16 obviously induced the expression of LC3 II and GFP-LC3 dots in MCF-7 cells, and decreased the expression of P62. Baf inhibited the au- tophagy induced by VP-16, and increased the expression of cleaved-PARP in MCF-7 cells and the apoptotic rates（P 〈 0. 01 ）. Conclusions VP-16 reduced the viability of breast cancer cells and induced protective autophagy, and inhibition of autophagy promoted the apoptotic death and increased the sensitivity of cells to VP-16.

关 键 词：依托泊苷 自噬 BAFILOMYCIN A1 凋亡 乳腺癌细胞 

分 类 号：R737.9[医药卫生—肿瘤]