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

发布时间：2018-09-19 12:00

【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。
[Abstract]:Objective to investigate the secretion and molecular mechanism of 17 尾 -estradiol (estradiol,E_2) -stimulated proliferation, migration and growth factor analogue of human clover factor 1 (trefoil factor family 1 (TFF1) in K-1 cells of papillary thyroid carcinoma. Methods the effect of E _ 2H _ 2ER 尾 agonist (propylpyrazoletriol,PPT) and ER 尾 agonist (diarylpropionitrile,DPN) on the secretion of TFF1 in K-1 cells was detected by ELISA. The expression of estrogen receptor ER 伪 -mer 尾 in K-1 cells was detected by Western blot. After ER 伪 siRNA,ER 尾 siRNA was synthesized and transfected into K-1 cells, ELISA was used to observe the effect of ER 伪 siRNA,ER 尾 siRNA on TFF1 secretion induced by E _ 2. Chromosome immunoprecipitation was used to detect the interaction between ER 伪 -ER 尾 and TFF1 gene promoter. The effect of E _ (2) on the proliferation of K-1 cells was detected by MTT and the effect of E _ (2) on the migration of K ~ (-1) cells was detected by Transwell. Results the content of TFF1 in K-1 cell supernatant increased gradually after treatment with EK 2, then reached its peak value at 24 h, then decreased after treated with PPT, the content of TFF1 increased, while DPN treatment decreased the content of TFF1, and the TFF1 secretion of K-1 cells decreased after transfection of ER 伪 siRNA. The expression of estrogen receptor ER 伪 in K-1 cells was higher than that of ER 尾. The results of chromosome immunoprecipitation showed that ER 伪 could bind to the promoter region of TFF1 gene. Conclusion EK _ 2 can promote the expression and secretion of TFF1 and the proliferation and migration of K-1 cells through ER 伪.
【作者单位】：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;
【基金】：国家自然科学基金面上项目(81272937)~~
【分类号】：R736.1

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