乳腺癌雌激素受体β不同亚型表达载体的构建及其转录活性分析  

Eukaryotic Expression Vectors Construction and Transcriptional Ac tivity Analysis of Different Isoforms of ERβ in Breast Cancer

作　　者：关鑫[1] 程龙[1] 邹大阳[1] 廉攀峰[2] 王超 叶棋浓[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850 [2]安徽医科大学附属安庆市立医院口腔科,安徽安庆246600 [3]北京王府中西医结合医院药剂科,北京102209

出　　处：《生物技术通讯》2013年第4期471-473,518,共4页Letters in Biotechnology

摘　　要：目的:构建人乳腺癌中雌激素受体β(ERβ)3种亚型ERβ1、ERβ2和ERβ5的真核表达载体,测定不同亚型ERβ的转录活性。方法:以乳腺癌细胞MCF7的cDNA为模板,PCR扩增ERβ1、ERβ2和ERβ5基因,分别克隆到pXJ40-Myc载体,Western印迹检测克隆载体在293T细胞内的表达;将上述载体与含雌激素应答元件(ERE)的萤光素酶(Luc)报告基因载体(ERE-luc)共转293T细胞,测定各亚型ERβ的转录活性。结果:构建了Myc-ERβ1、Myc-ERβ2和Myc-ERβ5表达载体,转录活性结果显示上述表达载体均具有活性,雌激素可升高ERβ1的转录活性,不能升高ERβ2和ERβ5的转录活性。结论:该研究为进一步探讨不同亚型ERβ在乳腺癌中的功能奠定了基础。Objective： To construct eukaryotic expression vectors and to detect transcriptional activities of three isoforms of human estrogen receptor β（ERβ） gene containing ERβ1, ERβ2 and ERβ5 expressed in breast cancer. Methods： ERβ1, ERβ2 and ERβ5 genes were amplified from eDNA of MCF7 breast cancer cells using PCR and inserted into the vector pXJ40-Myc. Western blot analysis was used to detect the expression of these isoforms. Transcriptional activities of ERfll, ERβ2 and ERβ5 were assayed by cotransfected these vectors and the luciferase reporter vector containing the estrogen-responsive element（ERE） into 293T cells. Results： Myc-ERβ1, Myc-ERβ2 and Myc-ERβ5 were successfully constructed as active forms. Estrogen could enhance activity of ERβ1 but not that of ERβ2 and ERβ5. Conclusion： This study could be a basis to further detect functions of different isoforms of ERβ in breast cancer.

关 键 词：雌激素受体Β 真核表达 转录活性 乳腺癌 

分 类 号：Q78[生物学—分子生物学]