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雌激素(E2)和雌激素受体(ER)在E2诱发的肿瘤中起着极其重要的作用。ER共调节因子通过与ER相互作用调节其生物学功能。PES1主要表达于E2的重要靶器官如乳腺、卵巢等组织中,并在乳腺癌细胞中高表达。用PCR技术构建HA标签的PES1全长以及1～322aa、312～588aa和414～588aa三个不同功能区片段的重组质粒。将不同的重组质粒与FLAG-ERα和或FLAGC-ERβ共转染293T细胞后进行免疫共沉淀,以验证PES1与ER是否有相互作用以及相互作用的区域。用含雌激素受体作用元件的荧光素酶报告基因(ERE-LUC)检测PES1对ERα和ERβ转录激活活性的影响。结果表明,PES1与ERα和ERβ均相互作用,且PES1的1～322aa区域与ERα和ERβ相结合。PES1能特异地、E2非依赖性抑制ERβ的转录激活活性。实验结果显示,PES1是一个新的ER共调节因子,需要进一步研究其在ERβ信号通路及其在E2诱发的肿瘤的作用。
Estrogen (E2) and estrogen receptor (ER) play an extremely important role in E2-induced tumors. ER co-regulatory factors regulate their biological function by interacting with ER. PES1 is mainly expressed in important target organs of E2 such as breast and ovary, and is highly expressed in breast cancer cells. The full-length PES1 with HA tag and three different functional region fragments of 1 ~ 322aa, 312 ~ 588aa and 414 ~ 588aa were constructed by PCR. 293T cells were cotransfected with different recombinant plasmids and FLAG-ERα and / or FLAGC-ERβ, then co-immunoprecipitated to verify whether PES1 and ER interact and interact with each other. The effect of PES1 on the transcriptional activation of ERα and ERβ was detected by luciferase reporter gene (ERE-LUC) containing estrogen receptor. The results showed that PES1 interacts with both ERα and ERβ, and the region from 1 to 322aa of PES1 binds to ERα and ERβ. PES1 specifically, E2-independent inhibit the transcriptional activation of ERβ. The experimental results show that PES1 is a new ER co-regulator, which needs further study of its role in ERβ signaling pathway and its role in E2-induced tumors.