ERα基因启动子甲基化状态对乳腺癌细胞系MTA1 mRNA和蛋白表达水平的影响

doi:10.13481/j.1671-587x.20150407

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (04): 706-710.doi: 10.13481/j.1671-587x.20150407

ERα基因启动子甲基化状态对乳腺癌细胞系MTA1 mRNA和蛋白表达水平的影响

孙亚楠¹, 毛晓韵², 刘崇², 陈浩², 郭扬², 金锋²

1. 中国医科大学附属第一医院普通外科教研室普外综合/烧伤科, 辽宁沈阳 110001;
2. 中国医科大学附属第一医院普通外科教研室乳腺外科, 辽宁沈阳 110001

收稿日期:2015-03-26 发布日期:2015-08-01
通讯作者: 金锋,教授,博士研究生导师(Tel:024-83282618,E-mail:jinfeng66cn@hotmail.com) E-mail:jinfeng66cn@hotmail.com
作者简介:孙亚楠(1982-),女,内蒙古自治区赤峰市人,主治医师,医学博士,主要从事乳腺癌发病的分子机制及手术治疗方面的研究。
基金资助:
国家自然科学基金资助课题(81201886)

Influence of estrogen receptor-alpha promoter methylation status in expression levels of MTA1 gene and protein in breast cancer cell lines

SUN Yanan¹, MAO Xiaoyun², LIU Chong², CHEN Hao², GUO Yang², JIN Feng²

1. Research Unit of General Surgery, Department of General Surgery & Burn Ward, First Affiliated Hospital, China Medical University, Shenyang 110001, China;
2. Research Unit of General Surgery, Department of Breast Surgery, First Affiliated Hospital, China Medical University, Shenyang 110001, China

Received:2015-03-26 Published:2015-08-01

摘要/Abstract

摘要：

目的: 探讨乳腺癌细胞系中雌激素受体α(ERα)启动子甲基化状态的改变对其MTA1 mRNA和蛋白表达水平的影响,阐明ERα启动子去甲基化对MTA1基因表达的作用。方法: 用去甲基化药物5-Aza-dC处理ERα启动子高甲基化状态的MDA-MB-435s和低甲基化状态的MDA-MB-231乳腺癌细胞系为5-Aza-dC处理组,同时设 2种细胞系的未处理组为对照组。甲基化特异性PCR(MSP)法检测各组细胞ERα启动子甲基化状态。Western blotting法检测各组细胞MTA1蛋白表达水平,RT-PCR法检测各组细胞MTA1 mRNA表达水平。结果: 5-Aza-dC逆转了MDA-MB-435s和MDA-MB-231细胞系ERα启动子甲基化状态; MDA-MB-435s乳腺癌细胞系中,与对照组比较,5-Aza-dC处理组MTA1 mRNA及蛋白表达水平均下调(P<0.05);MDA-MB-231乳腺癌细胞系中,与对照组比较,5-Aza-dC处理组MTA1 mRNA及蛋白表达水平亦均下调(P<0.05)。结论: 改变MDA-MB-435s和MDA-MB-231乳腺癌细胞系ERα基因启动子甲基化状态后,MTA1 mRNA和蛋白表达均下降,提示ERα启动子去甲基化后可以下调MTA1基因的表达,二者之间存在调控关系。

Abstract:

Objective To discuss the influence of the changes of estrogen receptor-alpha (ERα) promoter methylation in the expression levels of MTA1 mRNA and protein in breast cancer cell lines,and to illustrate the effect of ERα promoter on the expression of MTA1 gene after demethylation. Methods The breast cancer cell lines MDA-MB-435s in ERα promoter in hypermethylation status and MDA-MB-231 in ERα promoter in hypomethylation status were treated with 5-Aza-2'-deoxycytidine (5-Aza-dC) (5-Aza-dc treated groups),while the two kinds of cell lines were without 5-Aza-dC treatment were used as untreated groups.The methylation status of ERα promoter of the cells in various groups was detected by methylation-specific PCR (MSP).The expression levels of MTA1 mRNA and protein in MDA-MB-435s and MDA-MB-231 cells in various groups were detected with RT-PCR and Western blotting method. Results Both in the MDA-MB-435s and the MDA-MB-231 cells,the methylation status of ERα was modified from methylation to unmethylation by being treated with 5-Aza-dC.Furthermore,both in the MDA-MB-435s and the MDA-MB-231,compared with untreated groups,the expression levels of MTA1 mRNA and protein in 5-Aza-dC treated groups were down-regulated (P<0.05). Conclusion After the methylation status of ERα promoter is modified from methylation to unmethylation in breast cancer cell lines MDA-MB-435s and MDA-MB-231,the expression levels the MTA1 mRNA and protein are down-regulated.It's illustrated the expression of MTA1 can be lowered after ERα promoter demethylation,and the results further verify the regulation of the relationship between ERα methylation and MTA1 gene.

Key words: metastasis-associated gene 1, estrogen receptor-alpha, methylation, epigenetic, breast neoplasms

中图分类号:

R737.9

孙亚楠, 毛晓韵, 刘崇, 陈浩, 郭扬, 金锋. ERα基因启动子甲基化状态对乳腺癌细胞系MTA1 mRNA和蛋白表达水平的影响[J]. 吉林大学学报(医学版), 2015, 41(04): 706-710.

SUN Yanan, MAO Xiaoyun, LIU Chong, CHEN Hao, GUO Yang, JIN Feng. Influence of estrogen receptor-alpha promoter methylation status in expression levels of MTA1 gene and protein in breast cancer cell lines[J]. Journal of Jilin University Medicine Edition, 2015, 41(04): 706-710.

参考文献

[1] Toh Y,Pencil SD,Nicolson GL. A novel candidate metastasis-associated gene, mta1,differentially expressed in highly metastatic mammary adenocarcinoma cell lines. cDNA cloning,expression,and protein analyses [J].Biol Chem,1994(269):22958-22963.

[2] Wang RA.MTA1-a stress response protein:a master regulator of gene expression and cancer cell behavior [J].Cancer Metastasis Rev,2014,33(4):1001-1009.

[3] Vrtacnik P,Ostanek B,Mencej-Bedrac S,et al.The many faces of estrogen signaling [J].Biochem Med,2014,24(3):329-342.

[4] 孙亚楠,郭阿垚,毛晓韵,等.散发性乳腺癌中MTA1表达与ERα甲基化关系的研究[J].现代肿瘤医学,2013,21(6):1228-1231.

[5] Toh Y,Sugimachi K.The role of MTA1 gene expression in human hepatocellular carcinoma [J].Oncol Rep,2003,10(3):599-604.

[6] Toh Y,Ohga T,Endo K,et al.Expression of the metastasis-associated MTA1 protein and its relationship to deacetylation of the histone H4 in esophageal squamous cell carcinomas [J].Cancer,2004,110(3):362-367.

[7] 林川,陈汉,吴孟超,等.肿瘤转移基因MTAI在原发性肝癌中的表达及其临床意义[J].中华外科杂志,2000,38(12):915-917.

[8] 刘运贤,胡亚翎,张弦.肿瘤转移相关基因在肺癌的表达及其意义[J].蚌埠医学院学报,2009,34(5):421-422.

[9] Millard CJ,Fairall L,Schwabe JW.Towards an understanding of the structure and function of MTA1 [J].Cancer Metastasis Rev,2014,33(4):857-867.

[10] Nair SS,Li DQ,Kumar R.A core chromatin remodeling factor instructs global chromatin signaling through multivalent reading of nucleosome codes [J].Mol Cell,2013,49(4):704-718.

[11] Li DQ,Pakala SB,Nair SS,et al.Metastasis-associated protein 1/nucleosome remodeling and histone deacetylase complex in cancer [J].Cancer Res,2012,72(2):387-394.

[12] Li DQ,Kumar R.Mi-2/NuRD complex making inroads into DNA-damage response pathway [J].Cell Cycle,2010,9(11):2071-2079.

[13] Mazumdar A,Wang RA,Mishra SK,et al.Transcriptional repression of oestrogen receptor by metastasis-associated protein 1 corepressor [J].Nat Cell Biol,2001,3(1):30-37.

[14] Jiang Q,Zhang H,Zhang P.ShRNA-mediated gene silencing of MTA1 influenced on protein expression of ER alpha,MMP-9,CyclinD1 and invasiveness,proliferation in breast cancer cell lines MDA-MB-231 and MCF-7 in vitro [J].J Exp Clin Cancer Res,2011,30:60.doi:10.1186/1756-9966-30-60.

[15] Mirza S,Sharma G,Prasad CP,et al.Promoter hypermethylation of TMS1,BRCA1,ERalpha and PRB in serum and tumor DNA of invasive ductal breast carcinoma patients [J].Life Sci,2007,81(4):280-287.

[16] Nass SJ,Herman JG,Gabrielson E,et al.Aberrant methylation of the estrogen receptor and E-cadherin 5' CpG islands increases with malignant progression in human breast cancer [J].Cancer Res,2000,60(16):4346-4348.

[17] Sahab ZJ,Man YG,Semaan SM,et al.Alteration in protein expression in estrogen receptor alpha-negative human breast cancer tissues indicates a malignant and metastatic phenotype [J].Clin Exp Metastasis,2010,27(7):493-503.

[18] 陈洪岩,刘芝华.乳腺癌细胞雌激素受体非配体依赖性激活对来曲唑耐药的机制研究[J].解放军医学杂志,2013,38(6):461-466.

[19] 武萍萍,陶然,彭攸,等.EZH2及DNMT在三阴乳腺癌中的表达及意义[J].解放军医学杂志,2015,40(1):50-55.

ERα基因启动子甲基化状态对乳腺癌细胞系MTA1 mRNA和蛋白表达水平的影响

Influence of estrogen receptor-alpha promoter methylation status in expression levels of MTA1 gene and protein in breast cancer cell lines

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