乳腺癌C-erbB-2基因启动子区CpG岛的甲基化状态与雄激素受体表达的相关性分析

doi:10.3969/j.issn.1673-8640.2018.06.018

摘要/Abstract

摘要：

目的探讨乳腺癌中雄激素受体（AR）与人表皮生长因子受体2（C-erbB-2）基因启动子区CpG岛甲基化状态的关系,并明确AR与乳腺癌预后的关系。方法分别采用甲基化特异性聚合酶链反应（MSP）和免疫组织化学（IHC）方法分析76例乳腺癌患者癌组织和55例乳腺良性肿瘤患者肿瘤组织C-erbB-2基因启动子区CpG岛的甲基化状态和AR的表达水平,结合临床病理资料分析AR不同表达状态与C-erbB-2基因启动子区CpG岛甲基化水平的关系,并进行预后分析。结果 AR阳性的乳腺癌组织C-erbB-2基因启动子区CpG岛甲基化的比例[74.58%（44/59）]显著高于AR阴性的乳腺癌组织[47.06%（8/17）,P=0.032];C-erbB-2基因启动子区CpG岛的甲基化与AR蛋白的表达呈正相关（r=0.247,P=0.032）,且乳腺癌AR阳性患者无瘤生存率显著高于AR阴性患者（P<0.05）。结论乳腺癌组织AR蛋白的表达状态可作为乳腺癌预后转归的预测依据之一;C-erbB-2基因启动子区CpG岛的甲基化与AR蛋白的表达状态呈正相关,为乳腺癌的激素治疗及预后转归的预测提供了新的理论依据。

关键词: 雄激素受体, 人表皮生长因子受体2基因, 甲基化, 乳腺癌

Abstract:

Objective To investigate the correlation between androgen receptor（AR） and the methylation status of human epidermal growth factor receptor（C-erbB-2） gene promoter region CpG island in breast cancer,and to study the relationship between AR and clinical prognosis. Methods A total of 76 patients with breast cancer and 55 patients with benign breast tumor were enrolled,and the methylation status of C-erbB-2 gene promoter region CpG island and the expression level of AR were determined by methylation-specific polymerase chain reaction（MSP） and immunohistochemistry（IHC）,respectively. Based on clinical and pathological data,the relationship between AR and the methylation status of C-erbB-2 gene promoter region CpG island was evaluated,and the prognosis analysis was performed. Results The methylation proportion of C-erbB-2 gene promoter region CpG island in AR positive breast cancer tissue [74.58%（44/59）] was higher than that in AR negative breast cancer tissue [47.06%（8/17）]（P=0.032）. The methylation of C-erbB-2 gene promoter region CpG island was positively correlated with AR expression（r=0.247,P=0.032）,and the tumor free survival rate of AR positive patients was higher than that of AR negative patients（P<0.05）. Conclusions The expression status of AR in breast cancer tissue can be used as one of prognosis predictors. The methylation of C-erbB-2 gene promoter region CpG island is positively related to the expression status of AR,which provides a new reference for hormone therapy and breast cancer prognosis.

Key words: Androgen receptor, Human epidermal growth factor receptor gene, Methylation, Breast cancer

中图分类号:

R446.62

闫佩毅, 袁亚, 张骥, 朱琪, 邹玉涵, 王高莹, 金姝. 乳腺癌C-erbB-2基因启动子区CpG岛的甲基化状态与雄激素受体表达的相关性分析[J]. 检验医学, 2018, 33(6): 549-555.

YAN Peiyi, YUAN Ya, ZHANG Ji, ZHU Qi, ZOU Yuhan, WANG Gaoying, JIN Shu. Correlation between the methylation status of C-erbB-2 gene promoter region CpG island and androgen receptor expression in breast cancer[J]. Laboratory Medicine, 2018, 33(6): 549-555.

图/表 6

参考文献 20

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引物名称	序列（5'~3'）	产物大小（bp）
甲基化引物	MF AGGGAATTTATTTCGGATTTC	138
	MR CGCAAACCTAAATACGTCC
非甲基化引物	UF TTTAGGGAATTTATTTTGGATTTT	138
	UR CACAAACCTAAATACATCCCTC
野生型引物	WF GTGGACAGTCGAGACGCTCAG	440
	WR TGGTTGGGACAGGTTGCTAT

引物名称	序列（5'~3'）	产物大小（bp）
甲基化引物	MF AGGGAATTTATTTCGGATTTC	138
	MR CGCAAACCTAAATACGTCC
非甲基化引物	UF TTTAGGGAATTTATTTTGGATTTT	138
	UR CACAAACCTAAATACATCCCTC
野生型引物	WF GTGGACAGTCGAGACGCTCAG	440
	WR TGGTTGGGACAGGTTGCTAT

组别	例数	癌组织AR蛋白表达		χ²值	P值
组别	例数	阳性	阴性	χ²值	P值
C-erbB-2蛋白表达				1.376	0.241
阳性组	49	36	13
阴性组	27	23	4
C-erbB-2基因启动子区 CpG岛的甲基化				4.625	0.032
阳性组	52	44	8
阴性组	24	15	9
ER蛋白表达				2.250	0.134
阳性组	39	33	6
阴性组	37	26	11
PR蛋白表达				1.749	0.186
阳性组	33	28	5
阴性组	43	31	12
三阴性乳腺癌				0.406	0.524
三阴性乳腺癌组	8	5	3
非三阴性乳腺癌组	68	14	54
生存情况				1.605	0.205
存活组	67	54	13
死亡组	9	5	4
月经情况				0.350	0.554
绝经后组	61	46	15
绝经前组	15	13	2
癌灶浸润				1.217	0.569
导管内组	4	4	0
导管组	72	55	17

组别	例数	癌组织AR蛋白表达		χ²值	P值
组别	例数	阳性	阴性	χ²值	P值
C-erbB-2蛋白表达				1.376	0.241
阳性组	49	36	13
阴性组	27	23	4
C-erbB-2基因启动子区 CpG岛的甲基化				4.625	0.032
阳性组	52	44	8
阴性组	24	15	9
ER蛋白表达				2.250	0.134
阳性组	39	33	6
阴性组	37	26	11
PR蛋白表达				1.749	0.186
阳性组	33	28	5
阴性组	43	31	12
三阴性乳腺癌				0.406	0.524
三阴性乳腺癌组	8	5	3
非三阴性乳腺癌组	68	14	54
生存情况				1.605	0.205
存活组	67	54	13
死亡组	9	5	4
月经情况				0.350	0.554
绝经后组	61	46	15
绝经前组	15	13	2
癌灶浸润				1.217	0.569
导管内组	4	4	0
导管组	72	55	17

组别	例数	癌组织C-erbB-2 基因启动子区CpG岛的甲基化		χ²值	P值
组别	例数	甲基化	非甲基化	χ²值	P值
C-erbB-2蛋白表达				5.447	0.020
阳性组	49	29	20
阴性组	27	23	4
AR蛋白表达				4.625	0.032
阳性组	59	44	15
阴性组	17	8	9
ER蛋白表达				2.680	0.102
阳性组	39	30	9
阴性组	37	22	15
PR蛋白表达				0.501	0.479
阳性组	33	24	9
阴性组	43	28	15
生存情况				0.068	0.794
存活组	67	45	22
死亡组	9	7	2
月经情况				0.588	0.443
绝经前组	15	12	3
绝经后组	61	40	21
癌灶浸润				0.068	0.794
导管内组	4	2	2
导管组	72	50	22