17β-雌二醇促进甲状腺乳头状癌细胞增殖、迁移及三叶草因子1分泌  

17β-estradiol promotes proliferation,migration and TFF1 secretion in papillary thyroid cancer K-1 cells

作　　者：廖凌瑶 戴宇婕 赵婷婷[1] 刘智敏[1] 

机构地区：[1]重庆医科大学基础医学院生物化学与分子生物学教研室,分子医学与肿瘤研究中心,重庆400016

出　　处：《第三军医大学学报》2017年第17期1715-1719,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81272937)~~

摘　　要：目的研究17β-雌二醇(estradiol,E_2)刺激甲状腺乳头状癌K-1细胞增殖、迁移及生长因子类似物人三叶草因子1(trefoil factor family 1,TFF1)分泌及分子机制。方法 ELISA检测E_2、ERβ激动剂(propylpyrazoletriol,PPT)、ERβ激动剂(diarylpropionitrile,DPN)对K-1细胞TFF1分泌的影响,Western blot检测细胞中雌激素受体ERα、ERβ的表达量;设计合成ERαsiRNA、ERβsiRNA后,转染K-1细胞,采用ELISA观察其对E_2诱导的TFF1分泌的影响。染色体免疫共沉淀检测ERα、ERβ与TFF1基因启动子的相互作用;MTT检测E_2对K-1细胞增殖的影响;Transwell检测E_2对K-1细胞迁移的作用。结果 E_2处理后K-1细胞上清TFF1含量逐渐升高,24 h达到峰值,随后降低;PPT处理K-1细胞后TFF1含量增多,而DPN处理降低TFF1含量;转染ERαsiRNA后细胞TFF1分泌量减少;而转染ERβsiRNA后分泌量升高。Western blot检测结果显示,K-1细胞中雌激素受体ERα表达高于ERβ。染色体免疫共沉淀结果显示,ERα能与TFF1基因启动子区域结合;MTT结果显示,E_2处理促进K-1细胞增殖;Transwell检测结果显示,E_2处理增强K-1细胞迁移。结论 E_2可以通过ERα促进K-1细胞中TFF1的表达及分泌,促进细胞增殖和迁移。Objective To investigate whether 17β-estradiol （E2） can stimulate the proliferation, migration, and secretion of trefoil factor family 1 （ TFF1 ） in papillary thyroid cancer K-1 cells and explore the molecular mechanisms. Methods ELISA was used to detect the content of TFF1 in the supernatant of K-1 cells after the treatment of E2, propylpyrazoletriol （PPT, ERα agonist） or diarylpropionitrile （DPN, ERβ agonist）. The expression of ERct and ERα in the untreated cells was measured by Western blotting. ERα siRNA and ERβ siRNA by RNA interference were designed and synthesized, and the change of TFF1 was measured by ELISA again after the transfection. The interaction between TFF1 promoter and ER was evaluated by chromatin immunoprecipitation analysis （CHIP）. The proliferation and migration were detected in the K-1 cells after E2 treatment by MTT assay and Transwell chamber test respectively. Results After E2 treatment, the TFF1 content in the supernatant of K-1 cells was increased gradually, reached peak declined slowly. PPT treatment enhanced the secretion of TFF1 but DPN decreased it Transfection of ERα siRNA obliterated the inductive effect of E2 on the secretion of TFF1 siRNA increased the inductive effect in the K-1 cells. Western blotting showed that the at 24 h, and then in the K-1 cells. , but that of ERβ expression level ofERα was higher than that of ERβ in the K-1 cells. ChIP results confirmed that ERα protein was bound to the promoter of TFF1 gene in K-1 cells. E2 treatment promoted cell proliferation and improved cell migration in the K-1 cells. Conclusion E2 induces the expression and secretion of TFF1 in K-1 cells through ERα-dependent manner, and thus promotes the proliferation and migration of the cells.

关 键 词：17Β-雌二醇 甲状腺乳头状癌 三叶草因子1 雌激素受体 

分 类 号：R347.91[医药卫生—基础医学] R730.23