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  

Effects of silencing LSD1 and NDRG1 on migration of SKOV3 ovarian cancer cells

作　　者：邵根宝[1] 王冉冉 魏野[1] 金洁[1] 张柳平[1] 

机构地区：[1]江苏大学医学院,江苏镇江212013

出　　处：《山东医药》2017年第38期19-22,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81170573)

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P<0.01。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。Objective To observe the effects of silencing lysine specific demetylase 1（ LSD1） and N-myc downstream regulated gene 1（ NDRG1） on cell migration of ovarian cancer cells SKOV3 and to investigate the correlation between them.Methods Stable shRNA knockdown technique was employed to generate an inducible and stable SKOV3 cell line expressing tetracycline-regulated shRNA against LSD1（ LSD1-shRNA-SKOV3）.The LSD1-shRNA-SKOV3 cells were divided into the control group,Dox group,transfection group,and co-treatment group.Cells in the control group were treated with water,cells in the Dox group were treated with 100 ng/m L Dox,cells in the transfection group were transfected with NDRG1 siRNA,and cells in the co-treatment group were transfected with NDRG1 siRNA and were added with 100 ng/m L Dox at the same time.The mRNA and protein levels of LSD1 and NDRG1 genes in these cells were detected by realtime fluorescent quantitative PCR and Western blotting,respectively.The histone H3 lysine 4 dimethylation（ H3K4me2）levels in the promoter region of NDRG1 gene were measured by chromatin immunoprecipitation（ ChIP）.The cell migration rate was calculated by the Transwell chamber assay.Results The mRNA levels of LSD1 were 0.407 ± 0.029,0.936 ±0.024,0.413 ± 0.018,and 0.941 ± 0.035,and its protein levels were 0.306 ± 0.013,0.879 ± 0.036,0.341 ± 0.057,and 0.893 ± 0.052 in the Dox group,transfection group,co-treatment group,and control group,respectively.Compared with the control group,the mRNA and protein levels of LSD1 decreased in the Dox and co-treatment groups（ all P〈0.05）.The mRNA levels of NDRG1 were 0.791 ± 0.045,0.107 ± 0.016,0.165 ± 0.021,and 0.239 ± 0.027,and its protein levels were 0.907 ± 0.005,0.130 ± 0.006,0.216 ± 0.019,and 0.358 ± 0.062 in the Dox group,transfection group,co-treatment group,and control group,respectively.Compared with the control group,NDRG1 mRNA and protein levels significantly changed in the Dox group,transfection group and co-treatment group（ all P〈0.05）.T

关 键 词：赖氨酸特异性去甲基化酶1 N-myc下游调节基因1 表观遗传调控 细胞迁移 卵巢肿瘤 

分 类 号：R737.3[医药卫生—肿瘤]