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

doi:10.3969/j.issn.1004-616x.2017.06.003

癌变·畸变·突变 ›› 2017, Vol. 29 ›› Issue (6): 418-421,426.doi: 10.3969/j.issn.1004-616x.2017.06.003

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

肖红梅, 吴逸, 张婷, 王少康, 孙桂菊

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

收稿日期:2017-07-10 修回日期:2017-10-16 出版日期:2017-11-30 发布日期:2017-11-30
通讯作者: 王少康,E-mail:shaokangwang@seu.edu.cn E-mail:shaokangwang@seu.edu.cn
作者简介:肖红梅,E-mail:Cathyzh518@163.com。
基金资助:
国家自然科学基金项目（81372985，81673147）

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

XIAO Hongmei, WU Yi, ZHANG Ting, WANG Shaokang, SUN Guiju

Key Laboratory of Environmental Medicine and Engineering of Ministry of Education, School of Public Health, Southeast University, Nanjing 210009, Jiangsu, China

Received:2017-07-10 Revised:2017-10-16 Online:2017-11-30 Published:2017-11-30

摘要/Abstract

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、50 μg/mL）微囊藻毒素-LR处理人正常食管上皮HEEC细胞24 h，用四甲基偶氮唑蓝（MTT）法检测细胞活力，计算半数抑制浓度（IC₅₀）以确定后续实验的MC-LR浓度。进一步用0、6、12、24 μg/mL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，3.75~50 μg/mL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IC₅₀为24 μg/mL。因此，选择6、12和24 μg/mL的MC-LR用于后续实验。用6~24 μg/mL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。

关键词: 微囊藻毒素-LR, 人食管上皮细胞, 凋亡, Caspase-3, Caspase-9

Abstract: 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 (IC₅₀) which was used to determine the MC-LR concentrations for subsequent experiments. Accordingly,cells 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.

Key words: microcystin-LR, human esophageal epithelial cells, apoptosis, Caspase-3, Caspase-9

中图分类号:

R364

肖红梅, 吴逸, 张婷, 王少康, 孙桂菊. 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[J]. 癌变·畸变·突变, 2017, 29(6): 418-421,426.

XIAO Hongmei, WU Yi, ZHANG Ting, WANG Shaokang, SUN Guiju. Effects of microcystin-LR on apoptosis, and Caspase-3 and Caspase-9 expression in human normal esophageal epithelial cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(6): 418-421,426.

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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

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