敲低SETD8抑制乳腺癌MCF-7细胞的转移及其机制研究  

The study on the knockdown of SETD8 inhibiting the metastasis of breast cancer MCF-7 cells and its mechanism

作　　者：张晓静[1] 陈延松[1] 唐经纬 陈晨[1] 刘元 金功圣[1] 刘先富[1] Zhang Xiaojing;Chen Yansong;Tang Jingwei;Chen Chen;Liu Yuan;Jin Gongsheng;Liu Xianfu(Department of Surgical Oncology,The First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,Anhui,China)

机构地区：[1]蚌埠医学院第一附属医院肿瘤外科,安徽蚌埠233000

出　　处：《右江民族医学院学报》2022年第5期628-632,643,共6页Journal of Youjiang Medical University for Nationalities

基　　金：安徽省教育厅重点课题(KJ2019A0341);蚌埠医学院自然科学基金重点项目(2020byzd132)。

摘　　要：目的研究赖氨酸甲基转移酶(SETD8)对乳腺癌MCF-7细胞转移的调控作用。方法用qRT-PCR和免疫组化实验检测癌旁正常组织和乳腺癌组织中SETD8表达。使用Lipofectamine 3000分别将小干扰RNA阴性对照(Control),SETD8的小干扰RNA(siRNA-1、siRNA-2组)转染到MCF-7细胞中,Transwell实验检测细胞的迁移、侵袭能力。qRT-PCR检测SETD8、Slug、Zeb1、Zeb2、Twist1、Snail的mRNA表达量。Western blot检测SETD8和Slug的蛋白表达水平。染色质免疫共沉淀(ChIP)实验检测H4K20me1修饰在Slug基因启动子上的富集情况。结果与癌旁正常组织相比,乳腺癌组织中SETD8的mRNA和蛋白水平均显著升高(P<0.05)。与Control组比较,敲低组SETD8的mRNA和蛋白水平均显著降低(P<0.0001)。相比于Control组,细胞迁移、侵袭能力减弱,Slug蛋白表达水平降低。与IgG组比较,H4K20me1修饰能够显著富集在Slug基因启动子上游1500~2000 bp之间(P<0.001)。结论下调SETD8的表达能够抑制MCF-7细胞的转移能力,其机制可能与SETD8调控Slug基因转录相关。Objective To study the regulation of SETD8 in the metastasis of breast cancer MCF-7 cells.Methods The qRT-PCR and immunohistochemistry were used to detect the expression of SETD8 in normal tissues adjacent to the cancer and breast cancer tissues.The small interfering RNA of negative control group and SETD8 groups(siRNA-1,siRNA-2)was transfected into MCF-7 cells by Lipofectamine 3000,respectively.The ability of cell migration and invasion was tested by Transwell assay.The mRNA expression of SETD8,Slug,Zeb1,Zeb2,Twist1 and Snail were determined by qRT-PCR.The protein expression levels of SETD8 and Slug were tested by Western blot.Chromatin immunoprecipitation(ChIP)assay was used to detect the enrichment of H4K20me1 on Slug gene promoter.Results Compared with the adjacent normal tissues,the mRNA and protein levels of SETD8 were significantly increased in breast cancer tissues(P<0.05).The mRNA and protein levels of SETD8 in the knockdown group were significantly decreased compared with the control group(P<0.0001).Compared with the control group,the ability of cell migration and invasion was reduced,and the expression levels of Slug protein was decreased in the knockdown group.Compared with the IgG group,H4K20me1 was significantly enriched in the upstream of Slug gene promoter of 1500~2000 bp(P<0.001).Conclusion Down-regulation of SETD8 expression can inhibit the metastasis of MCF-7 cells,and the mechanism may be associated with the regulation of SETD8 in the Slug gene transcription.

关 键 词：乳腺肿瘤 赖氨酸甲基转移酶 MCF-7细胞 转移 

分 类 号：R737.9[医药卫生—肿瘤]