摘要
目的:利用RNA干扰技术靶向沉默EZH2基因,检测其对子宫内膜癌细胞增殖、迁移能力及上皮-间质转化(EMT)的影响。方法:实时荧光定量PCR、Western blot检测EZH2在子宫内膜细胞ESC及子宫内膜癌细胞株ECC-1、RL95-2中的表达差异。利用化学技术合成小分子siRNA转染子宫内膜癌细胞,靶向抑制EZH2基因表达。CCK-8、Transwell小室检测干扰前后子宫内膜癌细胞株ECC-1、RL95-2增殖、迁移能力变化。Western blot检测干扰前后子宫内膜癌细胞株ECC-1、RL95-2中EMT相关蛋白E-cadherin、α-catenin、N-cadherin和Vimentin的表达变化。结果:(1)EZH2在子宫内膜癌细胞株ECC-1、RL95-2中的表达明显高于子宫内膜细胞ESC(P<0.01)。(2)RNA干扰抑制EZH2基因表达后,ECC-1、RL95-2细胞增殖、迁移能力降低(P<0.01)。(3)RNA干扰抑制EZH2基因表达后,ECC-1、RL95-2细胞中上皮标志物E-cadherin和α-catenin的蛋白表达水平上调(P<0.05),间质标志物N-cadherin和Vimentin的蛋白表达水平下调(P<0.01)。结论:EZH2在子宫内膜癌细胞中高表达,siRNA靶向沉默EZH2基因可抑制子宫内膜癌细胞的增殖、迁移和上皮-间质转化。
Objective： To explore the effects on cell proliferation and migration as well as epithelial-mesenchymal transition（ EMT） after suppressing EZH2 expression in endometrial carcinoma.Methods： RT-PCR and Western blot were used to examine the expression of EZH2 in normal endometrial cell line ESC and endometrial carcinoma cell lines ECC-1 and RL95-2.SiRNAs targeting to EZH2 were transfected into endometrial carcinoma cells ECC-1 and RL95-2,CCK-8 and transwell migrating assay were utilized to examine the cell proliferation and migration.Western blot was applied to investigate the protein expression of E-cadherin、α-catenin、N-cadherin and Vimentin after siRNA targeting to EZH2 in endometrial carcinoma cell lines ECC-1 and RL95-2.Results： The expression of EZH2 in endometrial carcinoma cell lines ECC-1 and RL95-2 were higher than in normal endometrial cell line ESC（ P 0.01）. Further more,knocking down EZH2 expression could inhibit cell proliferation and migration in endometrial carcinoma cell lines ECC-1 and RL95-2（ P〈0.01）.Suppression of EZH2 could upregulated the expression of E- cadherin、α-catenin protein（ P〈0.05） and downregulated the expression of Ncadherin and Vimentin protein（ P〈0.01）.Conclusion： EZH2 expression is increased in endometrial carcinoma cell lines. Knocking down EZH2 expression suppresses the cell proliferation,migration and epithelial-mesenchymal transition in endometrial carcinoma cells.
出处
《现代妇产科进展》
CSCD
北大核心
2017年第2期99-103,共5页
Progress in Obstetrics and Gynecology
基金
上海市科委基金资助项目(No:20134226)
关键词
子宫内膜癌
EZH2基因
增殖
迁移
上皮-间质转化
Endometrial carcinoma
EZH2 gene
Proliferation
Migration
Epithelial-mesenchymal transition（EMT）
