米非司酮通过调控H19基因甲基化抑制子宫内膜癌细胞的迁移  

Mifepristone inhibites the migration of endometrial cancer cells through regulating H19 methylation

作　　者：路真真 颜磊[1] 张辉[1] 李明江[1] 张晓晖[1] 赵兴波[1] 

机构地区：[1]山东大学附属省立医院妇产科,济南250021

出　　处：《中华肿瘤杂志》2016年第6期411-416,共6页Chinese Journal of Oncology

基　　金：国家自然科学基金（81272858、81202057、81571414）

摘　　要：目的探讨米非司酮对子宫内膜癌Ishikawa细胞迁移的影响及其作用机制。方法体外培养人子宫内膜癌Ishikawa细胞。以不同浓度的米非司酮处理Ishikawa细胞，采用细胞划痕实验观察米非司酮对Ishikawa细胞迁移的影响，采用逆转录聚合酶链反应和甲基化特异性聚合酶链反应检测Ishikawa细胞中I-t19mRNA的表达及其甲基化变化，采用Westernblot法检测Ishikawa细胞中H19的下游基因高迁移率族蛋白A2（HMGA2）的表达情况以及相关的上皮一间质转化蛋白的表达。结果米非司酮处理48h时，对照组、5mg／L米非司酮处理组、10mg／L米非司酮处理组Ishikawa细胞的划痕宽度分别为（4．18±0．07）mm、（4．68±0．07）mm和（4．99±0．07）mm；与对照组比较，各药物处理组差异均有统计学意义（均P〈0．05）。处理72h时，对照组、5mg／L米非司酮处理组、10mg／L米非司酮处理组Ishikawa细胞的划痕宽度分别为（3．46±0．07）mm、（4．29±0．07）mm和（4．78±0．04）mm；与对照组比较．各药物处理组差异均有统计学意义（均P〈0．05）。随着处理时间和浓度的增加，米非司酮对Ishikawa细胞中H19mRNA表达的抑制作用增加，对H19基因启动子甲基化的促进作用增强，对HMGA2蛋白表达的抑制作用增强，从而使E．cadherin蛋白表达增加，Vimentin蛋白表达减少。结论米非司酮能通过上调H19基因甲基化下调H19mRNA表达，进而下调HMGA2和Vimentin蛋白表达，上调E—cadherin表达．抑制子宫内膜癌细胞迁移。Objective To investigate the effect and mechanism of mifepristone on the migration of human endometrial carcinoma cells. Methods A human endometrial carcinoma cell line, Ishikawa cells, was cultured in vitro and treated with mifepristone at different concentrations. Wound healing assay was applied to detect the migration of Ishikawa cells. RT-PCR and methylation-specific PCR （MSP） were used to detect the levels of H19 mRNA and its DNA methylation. Western-blot was used to detect the expressions of HMGA2 and epithelial to mesenchymal transition （EMT） related proteins. Results When treated with different concentrations of mifepristone for 48 hours, the width of scratch of the the control group, the 5 mg/L and the 10 mg/L mifepristone treatment groups were （4.18±0.07） mm, （4.68±0.07） mm, and （4.99±0.07） mm, respectively （P〈0.05 for all） and treated with mifepristone for 72 hours, those were（3.46±0.07）mm, （4.29±0.07） mm, and（4.78±0.04） mm, respectively （ P〈0.05 for all）. In the Ishikawa cells, mifepristone suppressed the transcriptional level of H19 through enhancing its promoter methylation, which resulted in inhibited expressions of HMGA2 and vimentin and increased expression of E-cadherin in a time- and concentration-dependent manner. Conclusion Mifepristone inhibits the migration of endometrial carcinoma cells partially through methylation-induced of transcriptional inhibition of H19, which results in the down- regulation of HMGA2 and vimentin and upregulation of E-cadherin.

关 键 词：子宫内膜肿瘤 甲基化 米非司酮 H19基因 

分 类 号：R737.33[医药卫生—肿瘤]