RNA干扰抑制EZH2基因表达对子宫内膜癌细胞上皮-间质转化的影响

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(P0.01)。(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(P0.01)。(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(P0.05),间质标志物N-cadherin和Vimentin的蛋白表达水平下调(P0.01)。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。

Influence of inhibition of EZH2 gene expression by RNA interference on epithelial-mes-enchymal transition in endometrial carcinoma cells

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 mi-gration. Western blot was applied to investigate the protein expression of E- cadherin、α-cate-nin、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 endome-trial carcinoma cell lines ECC-1 and RL95-2 ( P<0 . 01 ) . Suppression of EZH2 could upregulat-ed the expression of E-cadherin、α-catenin protein(P<0. 05) and downregulated the expression of N-cadherin and Vimentin protein ( P<0 . 01 ) . Conclusion:EZH2 expression is increased in endometrial carcinoma cell lines. Knocking down EZH2 expression suppresses the cell prolifera-tion,migration and epithelial-mesenchymal transition in endometrial carcinoma cells.