CAJ | 학술논문

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。
목적：구건인유선암중자격소수체β（ERβ）3충아형ERβ1、ERβ2화ERβ5적진핵표체재체，측정불동아형ERβ적전록활성。방법：이유선암세포MCF7적cDNA위모판，PCR확증ERβ1、ERβ2화ERβ5기인，분별극륭도pXJ40-Myc재체，Western인적검측극륭재체재293T세포내적표체；장상술재체여함자격소응답원건（ERE）적형광소매（Luc）보고기인재체（ERE-luc）공전293T세포，측정각아형ERβ적전록활성。결과：구건료Myc-ERβ1、Myc-ERβ2화Myc-ERβ5표체재체，전록활성결과현시상술표체재체균구유활성，자격소가승고ERβ1적전록활성，불능승고ERβ2화ERβ5적전록활성。결론：해연구위진일보탐토불동아형ERβ재유선암중적공능전정료기출。
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 cDNA 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 ERβ1, 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.