LSD1、NDRG1基因沉默对人卵巢癌SKOV3细胞迁移能力的影响及两者的关系

发布时间：2018-05-06 09:36

  本文选题：赖氨酸特异性去甲基化酶 + N-myc下游调节基因　； 参考：《山东医药》2017年38期

【摘要】：目的观察赖氨酸特异性去甲基化酶1(LSD1)、N-myc下游调节基因1(NDRG1)基因沉默对人卵巢癌SKOV3细胞迁移能力的影响,并探讨两者的作用关系。方法取人卵巢癌SKOV3细胞株,通过shRNA方法建立诱导型干扰LSD1的SKOV3细胞株(LSD1-shRNA-SKOV3),将其分为对照组、Dox组、转染组和联合组。对照组用水处理,Dox组用100 ng/m L Dox处理,转染组转染NDRG1 siRNA,联合组转染NDRG1 siRNA的同时加入100 ng/m L Dox处理。采用实时荧光定量PCR和Western blotting法分别检测4组LSD1、NDRG1基因mRNA和蛋白表达;染色质免疫沉淀ChIP法和实时荧光定量PCR法分析对照组和Dox组NDRG1基因启动子区组蛋白H3第4位赖氨酸的二甲基化(H3K4me2)程度;Transwell小室测算4组细胞迁移率。结果 Dox组、转染组、联合组、对照组LSD1mRNA表达分别为0.407±0.029、0.936±0.024、0.413±0.018、0.941±0.035,蛋白表达分别为0.306±0.013、0.879±0.036、0.341±0.057、0.893±0.052,Dox组、联合组与对照组相比,P均0.05。Dox组、转染组、联合组、对照组NDRG1 mRNA表达分别为0.791±0.045、0.107±0.016、0.165±0.021、0.239±0.027,蛋白表达分别为0.907±0.005、0.130±0.006、0.216±0.019、0.358±0.062,Dox组、转染组、联合组与对照组相比,联合组与Dox组、转染组相比,P均0.05。Dox组、对照组细胞NDRG1基因启动子区H3K4me2水平分别为3.32±0.41、0.83±0.17,两组相比P0.01。Dox组、转染组、联合组、对照组细胞迁移率分别为21.75%±1.816%、79.13%±2.561%、40.13%±2.039%、68.91%±3.167%,Dox组、转染组、联合组与对照组相比,联合组与Dox组、转染组相比,P均0.05。结论 LSD1基因沉默人卵巢癌SKOV3细胞迁移能力降低,NDRG1基因沉默人卵巢癌SKOV3细胞迁移能力升高;LSD1通过降低NDRG1基因启动子区域H3K4me2水平,抑制NDRG1的表达,从而促进卵巢癌SKOV3细胞转移。
[Abstract]:Objective to investigate the effect of Lysine-specific demethylase (LSD1) -N-myc gene silencing on the migration of human ovarian cancer SKOV3 cells. Methods Human ovarian cancer SKOV3 cell line was selected and induced LSD1 interference SKOV3 cell line (LSD1-shRNA-SKOV3) was established by shRNA. The cells were divided into three groups: control group, transfection group and combined group. The control group was treated with 100 ng/m L Dox, the control group was treated with 100 ng/m L Dox, and the combined group was treated with NDRG1 siRNA and 100 ng/m L Dox. Real-time fluorescence quantitative PCR and Western blotting were used to detect the mRNA and protein expression of NDRG1 gene in four groups. Chromatin immunoprecipitation (ChIP) and real-time fluorescence quantitative PCR (PCR) were used to analyze the level of dimethylated H3K4me2 of NDRG1 gene promoter H3 in control group and Dox group. 缁撴灉 Dox缁，

本文编号：1851805