微囊藻毒素-LR对人正常食管上皮细胞凋亡及对Caspase-3和Caspase-9蛋白表达的影响  

Effects of microcystin-LR on apoptosis, and Caspase-3 and Caspase-9 expression in human normal esophageal epithelial cells

作　　者：肖红梅 吴逸 张婷 王少康 孙桂菊 

机构地区：[1]东南大学公共卫生学院环境医学工程教育部重点实验室,江苏南京210009

出　　处：《癌变．畸变．突变》2017年第6期418-421,426,共5页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金项目(81372985;81673147)

摘　　要：目的:研究微囊藻毒素-LR对人正常食管上皮细胞活力及凋亡相关蛋白Caspase-3和Caspase-9表达的影响,为探讨其诱导凋亡的途径提供依据。方法:分别用不同浓度(1、3.75、7.5、15、30、50μg/m L)微囊藻毒素-LR处理人正常食管上皮HEEC细胞24h,用四甲基偶氮唑蓝(MTT)法检测细胞活力,计算半数抑制浓度(IC)以确定后续实验的MC-LR浓度。进一步用0、6、12、24 50μg/m L微囊藻毒素-LR处理细胞,采用PI-Annexin V双染法检测细胞凋亡,Western blot检测Caspase-3和Caspase-9蛋白的表达水平。结果:与对照组比较,3.75~50μg/m L的微囊藻毒素-LR均可使人正常食管上皮细胞活力降低(P均<0.01),测定得出IC为24 50μg/m L。因此,选择6、12和24μg/m L的MC-LR用于后续实验。用6~24μg/m L微囊藻毒素-LR处理细胞24 h后,细胞凋亡率升高(P<0.05或P<0.01),Caspase-3和Caspase-9蛋白表达增加(P均<0.01)。结论:微囊藻毒素-LR可以诱导人正常食管上皮细胞凋亡,且可能与细胞凋亡相关的信号分子Caspase-3和Caspase-9的激活有关。OBJECTIVE：To investigate the effects of microcystin-LR on viability and apoptosis of human normal esophageal epithelial cells,and on expression of Caspase-3 and Caspase-9. METHODS：Human normal esophageal epithelial cells were treated with microcystin-LR at different concentrations （1,3.75,7.5,15,30,50 μg/mL） for 24 h,and cell viability was detected by the MTT assay. The data were used to calculate the half inhibitory concentration （IC ） which was used to determine the MC-LR concentrations for subsequent experiments. Accordingly, 50cells were treated with 6,12 and 24 μg/mL MC-LR,and apoptosis was detected by PI-Annexin V double staining. Expression levels of Caspase-3 and Caspase-9 were detected by Western blot. RESULTS：Compared with the control group,microcystin-LR of 3.75-50 μg/mL decreased the viability of normal esophageal epithelial cells （P〈0.01）;the 6-24 μg/mL concentrations increased the apoptosis rate （P〈0.05 or P〈0.01）,and the expression of Caspase-3 and Caspase-9 （P〈0.01）. CONCLUSION：Induction of apoptosis in human normal esophageal epithelial cells by microcystin-LR may be related to its activation of Caspase-3 and Caspase-9.

关 键 词：微囊藻毒素-LR 人食管上皮细胞 凋亡 CASPASE-3 CASPASE-9 

分 类 号：R364[医药卫生—病理学]