雌激素上调TFF3促进甲状腺乳头状癌细胞增殖的研究  

Estrogen Enhances Papillary Thyroid Cancer Cell Proliferation by Upregulation of TFF3

作　　者：张文静[1] 张辰露 余鸿杰 李雪 王佳轩 张静[1] 王冬梅[1] 林旭 ZHANG Wen-jing;ZHANG Chen-lu;YU Hong-jie;LI Xue;WANG Jia-xuan;ZHANG Jing;WANG Dong-mei;LIN Xu(Morphology Laboratory of Hebei North University,Zhangjiakou Key Laboratory,Zhangjiakou,Hebei 075000,China)

机构地区：[1]河北北方学院形态学实验室张家口市重点实验室,河北张家口075000

出　　处：《河北北方学院学报（自然科学版）》2021年第10期1-7,共7页Journal of Hebei North University:Natural Science Edition

基　　金：河北省卫生厅科技计划(No.20170861);河北北方学院大学生创新计划项目(No.xj202040);河北省教育厅青年基金项目(No.QN2017011);河北省卫生厅科技项目(No.20170860);张家口市科学技术项目(No.1821030D)。

摘　　要：目的探讨雌激素与三叶因子3(trefoil factor family 3,TFF3)在甲状腺乳头状癌(papillary thyroid carcinoma,PTC)癌细胞增殖中的作用。方法免疫组织化学法检测雌激素受体α(estrogen receptorα,ERα)在PTC组织和癌旁组织中的表达及其与临床病理参数的关系;不同浓度雌激素刺激甲状腺乳头状癌K1细胞24 h后,采用实时荧光定量PCR(real-time quantitative PCR)和蛋白质印迹(Western blot)法检测TFF3基因和蛋白水平变化,MTT法检测K1细胞增殖率;ER抑制剂ICI182780与10 nM雌激素作用K1细胞后,Western blot与MTT法分别检测TFF3蛋白水平和细胞增殖活力的变化;双荧光素酶报告基因检测不同浓度雌激素作用K1细胞24 h后TFF3荧光素酶活性的变化;qPCR检测TFF1、Bcl-2、cyclin D mRNA水平变化。结果PTC组织中ERα阳性表达率高于癌旁组织(P<0.05),与性别、临床分期、淋巴结转移、TFF3阳性表达率具有相关性(P<0.05)。TFF3 mRNA及蛋白水平、K1细胞增殖率随雌激素浓度升高而增高(P<0.01);ICI182780能够抑制雌激素作用下K1细胞TFF3蛋白水平及增殖活力的增高(P<0.01)。雌激素显著提高了K1细胞TFF3荧光素酶活性,并提高K1细胞雌激素反应基因TFF1、抗凋亡基因Bcl-2、细胞周期基因cyclin D mRNA水平(P<0.01)。结论雌激素信号通路在PTC中促进了TFF3表达,进而促进了PTC癌细胞的增殖。Objective To investigate the role of estrogen and trefoil factor family 3(TFF3)in cancer cell proliferation of papillary thyroid carcinoma(PTC).Methods The expression of estrogen receptorα(ERα)in a PTC tissue microarray containing PTC and paracancerous tissues was detected by immunohistochemistry and its relationship with clinicopathological parameters was further analyzed;after stimulating K1 cells with different concentration of estrogen for 24 hours,the expression levels of TFF3 were detected by real-time quantitative PCR(qPCR)and Western blot,and the proliferation rate of K1 cells was detected by MTT.Western blot and MTT assay were respectively used to detect the changes of TFF3 protein level and cell proliferation activity in K1 cells treated with ICI182780(a reagent for inhibiting estrogen)and 10 nM estrogen.Double luciferase report gene experiment was used to detect the changes of TFF3 luciferase activity in K1 cells treated with different concentrations of estrogen for 24 hours.The mRNA levels of TFF1,Bcl-2 and cyclin D were detected by qPCR.Results The expression of ERαin PTC tissue was significantly higher than that in paracancerous tissue(P<0.05),which was correlated with gender,clinical stage,lymph node metastasis and TFF3 positive expression rate(P<0.05).TFF3 mRNA and protein levels and K1 cell proliferation rate increased with the increase of estrogen concentration(P<0.01).ICI182780 could inhibit the increase of TFF3 protein level and proliferation activity in K1 cells induced by estrogen(P<0.01).Estrogen significantly increased the luciferase activity of TFF3 in K1 cells,and promoted the transcription of estrogen responsive gene TFF1,anti-apoptotic gene Bcl-2 and cell cycle gene cyclin D(P<0.01).Conclusion Estrogen signaling pathway promotes the up-regulation of TFF3 expression in PTC and further promotes the cancer cell proliferation of PTC.

关 键 词：雌激素 三叶因子3 甲状腺乳头状癌 癌细胞增殖 

分 类 号：R73-3[医药卫生—肿瘤]