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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 study the effects of 17β-estradiol (E_2) on the proliferation and migration of thyroid papillary carcinoma K-1 cells and the molecular mechanism of trefoil factor family 1 (TFF1) secretion. Methods The effect of E_2, PPT and DPN on the secretion of TFF1 in K-1 cells was detected by ELISA. The expressions of ERα and ERβ in esophageal cancer cells were detected by Western blot. The ERαsiRNA and ERβsiRNA were transfected into K-1 cells and their effects on E_2 -induced TFF1 secretion were observed by ELISA. Chromosome co-immunoprecipitation was used to detect the interaction of ERα, ERβ and TFF1 promoter; MTT was used to detect the effect of E_2 on the proliferation of K-1 cells; Transwell was used to detect the effect of E_2 on the migration of K-1 cells. Results The content of TFF1 in the supernatant of K-1 cells increased gradually after E_2 treatment and peaked at 24 h, and then decreased. The content of TFF1 in K-1 cells increased after PPT treatment, but decreased in TDP1 treated with DPN. Decreased; while ERβsiRNA transfection increased secretion. Western blot results showed that the expression of estrogen receptor ERα in K-1 cells was higher than that of ERβ. Chromosome co-immunoprecipitation showed that ERα could bind to the promoter region of TFF1 gene; MTT assay showed that E_2 treatment promoted the proliferation of K-1 cells; Transwell assay showed that E_2 treatment enhanced the migration of K-1 cells. Conclusion E 2 can promote the expression and secretion of TFF1 in K-1 cells through ERα, and promote cell proliferation and migration.