DRD2启动子低甲基化通过ERK通路调控乳腺癌细胞增殖  

Hypomethylation of DRD2 promoter modulated breast cancer cells proliferation through ERK pathway

作　　者：钟明 袁浩 钱丰源 朱红波 伍雯 蒋晓飞 游庆华 李永平 ZHONG Ming;YUAN Hao;QIAN Feng-yuan;ZHU Hong-bo;WU Wen;JIANG Xiao-fei;YOU Qing-hua;LI Yong-ping(Department of Breast Surgery,Shanghai Pudong Hospital-Fudan University Pudong Medical Center,Shanghai 201399,China;Department of Pathology,Shanghai Pudong Hospital-Fudan University Pudong Medical Center,Shanghai 201399,China)

机构地区：[1]上海市浦东医院-复旦大学附属浦东医院乳腺外科,上海201399 [2]上海市浦东医院-复旦大学附属浦东医院病理科,上海201399

出　　处：《复旦学报（医学版）》2022年第5期704-712,719,共10页Fudan University Journal of Medical Sciences

基　　金：上海市浦东新区卫生系统重点专科建设项目(PWZzk2017-32)。

摘　　要：目的研究肿瘤干细胞标志物多巴胺受体D2(dopamine receptor D2,DRD2)对乳腺癌细胞增殖的影响。方法采用850K芯片对乳腺癌组织进行检测,并分析DRD2甲基化状态。采用基因筛选、基因敲除和甲基化抑制等技术和方法,进行体外和体内实验,筛选和验证可能存在的分子信号通路。结果DRD2启动子区在乳腺癌组织中相较于正常组织表现为低甲基化。上调DRD2的表达后,乳腺癌细胞增殖增强,而下调DRD2的表达,乳腺癌细胞增殖显著降低。裸鼠实验发现,过表达DRD2可促进肿瘤生长和Ki67、CD31表达,下调DRD2可抑制肿瘤生长。体内外实验表明,细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)的表达受DRD2表达水平的影响,提示DRD2通过ERK信号通路发挥作用。甲基化抑制剂5-aza-2-脱氧胞苷(5-aza-2-deoxycytidine,5-AzadC)在小鼠体内部分逆转了DRD2表达的下调,失去了对肿瘤细胞增殖的抑制作用,提示抑制DRD2甲基化促进了肿瘤的发展。结论乳腺癌中DRD2启动子区发生低甲基化。DRD2通过ERK通路在乳腺癌细胞的增殖和迁移中发挥作用。Objective To investigate the effect of tumor stem cell marker dopamine receptor D2(DRD2)on the proliferation of breast cancer cells Methods The breast cancer tissue was detected by 850K chip and the status of DRD2 methylation was analyzed.Experiments were carried out in vitro and in vivo to detect and verify the possible molecular signaling pathways by the techniques and methods of gene screening,gene knockout and methylation inhibition.Results DRD2 promoter region was hypomethylated in breast cancer tissues compared with normal tissues.After up regulation of DRD2expression,the proliferation of breast cancer cells was enhanced.While down regulation of DRD2expression,the proliferation of breast cancer cells was significantly reduced.Nude mice experiments found that overexpression of DRD2 could promote tumor growth and expression of Ki67 and CD31,while downregulation of DRD2 could inhibit tumor growth.Experiments showed that extracellular regulated protein kinases(ERK)expression was affected by the expression level of DRD2 in vitro and in vivo,suggesting that DRD2 functions through the ERK signaling pathway.Methylation inhibitor 5-aza-2-deoxycytidine(5-AzadC)partially reversed the inhibitory effect of DRD2 down-expression on cell proliferation and migration,as well as that on tumor growth in mice,which indicated that inhibition of DRD2 methylation can promote cancer development.Conclusion The promoter region of DRD2 gene is hypomethylated in breast cancer.DRD2 played a role in the proliferation and migration of breast cancer cells through ERK pathway.

关 键 词：乳腺癌 多巴胺受体D2(DRD2) 甲基化 细胞外调节蛋白激酶(ERK)通路 

分 类 号：R737.9[医药卫生—肿瘤]