SOX7基因启动子甲基化水平对乳腺癌MDA-MB-231细胞株体外迁移和侵袭的影响  

Effect of SOX7 Promoter Methylation Level on Migration and Invasion of Breast Cancer MDA-MB-231 Cells Line in Vitro

作　　者：王昂[1] 王杰 鄢文[1] 关小倩 廖明 WANG Ang;WANG Jie;YAN Wen;GUANXiao-qian;LIAO Min(The First Ward of Oncology,Guangdong No.2 Provincial People's Hospital,Guangzhou,Guangdong,510317,China;Department of Radiotherapy,Guangdong No.2 Provincial People's Hospital,Guangzhou,Guangdong,510317,China;Department of Oncology,The Third Affiliated Hospital of Southern Medical University,Guangzhou,Guangdong,510000,China;Department of Thoracic Surgery,General Hospital of Southern War Zone of Chinese People's Liberation Army,Guangzhou,Guangdong,510010,China)

机构地区：[1]广东省第二人民医院肿瘤一区,广东广州510317 [2]广东省第二人民医院放疗科,广东广州510317 [3]南方医科大学第三附属医院肿瘤内科,广东广州510000 [4]中国人民解放军南部战区总医院胸外科,广东广州510010

出　　处：《现代生物医学进展》2019年第22期4238-4242,共5页Progress in Modern Biomedicine

基　　金：广东省自然科学基金项目(2018A0303130050)

摘　　要：目的:探讨乳腺癌MDA-MB-231细胞中,Y性别决定区基因7(SOX7)基因启动子甲基化水平对细胞的体外迁移和侵袭的影响。方法:脂质体转染pcDNA3.0-DNA甲基转移酶3a(DNMT3a)质粒至MDA-MB-231细胞中,并于24h、48h及72h后,采用蛋白质免疫印迹实验(WB)检测细胞内DNMT3a蛋白表达水平;甲基化特异性定量PCR(Q-MSP)检测DNMT3a处理组、5-aza-C处理组及对照(Control)组MDA-MB-231细胞中的SOX7基因启动子DNA甲基化水平;实时荧光定量PCR(qRT-PCR)及WB实验检测各组MDA-MB-231细胞中的SOX7 m RNA和蛋白表达水平;细胞划痕实验及细胞侵袭实验检测各组MDA-MB-231细胞的迁移和侵袭能力。结果:pcDNA3.0-DNMT3a质粒转染MDA-MB-231细胞24h时,细胞内的DNMT3a蛋白表达水平最高。DNMT3a能够显著提高SOX7基因启动子DNA甲基化水平,而5-aza-C则抑制了SOX7基因启动子DNA甲基化水平(P<0.05)。与Control组相比,DNMT3a处理组的MDA-MB-231细胞中,SOX7的m RNA及蛋白表达水平均明显下降,而5-aza-C处理组SOX7的m RNA及蛋白表达水平均明显增加(P<0.05)。与Control组相比,DNMT3a处理组的MDA-MB-231细胞的迁移和侵袭能力均显著增强(P<0.05),而5-aza-C处理组的MDA-MB-231细胞的迁移和侵袭能力变化不大(P>0.05)。结论:在恶性肿瘤中,SOX7低表达表受其基因启动子高甲基化调节,且乳腺癌MDA-MB-231细胞中低表达的SOX7能够影响细胞的外迁移和侵袭能力。Objective: To investigate the effect of promoter methylation level of sex determining region Y-box 7(SOX7) gene on cells migration and invasion in vitro in breast cancer MDA-MB-231 cells. Methods: Liposome transfection of pcDNA3.0-DNA methylation transferase 3a(DNMT3a) plasmid into MDA-MB-231 cells, after 24 h, 48 h and 72 h, the expression of DNMT3a protein in cells was detected by Western blotting(WB). DNA methylation level of SOX7 gene promoter in MDA-MB-231 cells in DNMT3a treatment group, 5-aza-C treatment group and control groups were detected by quantitative methylation specific PCR(Q-MSP). The expression of SOX7 in MDA-MB-231 cells were detected by real-time quantitative PCR(qRT-PCR) and WB assay. Detection of migration and invasion energy of MDA-MB-231 cells by cell scratch test and cell invasion test in each groups. Results: The highest expression level of DNMT3a protein was found in MDA-MB-231 cells transfected with pcDNA3.0-DNMT3 a plasmid for 24 hours.DNMT3a significantly increased the DNA methylation level of SOX7 promoter, while 5-aza-C inhibited the DNA methylation level of SOX7 promoter(P<0.05). Compared with the control group, the expression of SOX7 in MDA-MB-231 cells treated with DNMT3a decreased significantly, while the expression of SOX7 in 5-aza-C group increased significantly(P<0.05). Compared with the control group, the migration and invasion ability of MDA-MB-231 cells treated with DNMT3a were significantly enhanced(P<0.05), while the migration and invasion ability of MDA-MB-231 cells treated with 5-aza-C did not change significantly(P>0.05). Conclusion: In malignant tumors, the low expression of SOX7 is regulated by the hypermethylation of SOX7 promoter, and the silencing of SOX7 significantly influence the migration and invasion of breast cancer MDA-MB-231 cells line in vitro.

关 键 词：SOX7 甲基化水平 MDA-MB-231 迁移 侵袭 

分 类 号：R-33[医药卫生] R737.9