基础医学与临床 ›› 2016, Vol. 36 ›› Issue (7): 918-923.

• 研究论文 • 上一篇    下一篇

镉通过GPER1-ERK/AKT通路促进甲状腺癌WRO细胞系增殖

朱平1,莫小梅2,刘智敏2   

  1. 1. 重庆医科大学
    2. 重庆医科大学基础医学院生物化学与分子生物学教研室
  • 收稿日期:2015-12-28 修回日期:2016-04-28 出版日期:2016-07-05 发布日期:2016-06-22
  • 通讯作者: 刘智敏 E-mail:liuzm9999@aliyun.com
  • 基金资助:
    甲状腺癌细胞雌激素结合蛋白及其相互作用分子的研究

Cadmium promotes proliferation of thyroid cancer cell line WRO through GPER1-ERK/AKT pathway

  • Received:2015-12-28 Revised:2016-04-28 Online:2016-07-05 Published:2016-06-22

摘要: 目的检测镉对甲状腺癌WRO细胞系的增殖作用,及G蛋白偶联雌激素受体1(GPER1、GPR30)在该过程中的作用。方法Western blot检测MCF-7、MDA-MB-231及WRO 3种细胞系GPER1的表达;将3种细胞分别分为7组进行处理:空白对照组、镉(Cd)(250,500,750及1000 nmol/L )处理组、17β-雌二醇(E2)(10 nmol/L)处理组和GPER1特异性激动剂(G1)(10 nmol/L)处理组,MTT法检测细胞增殖;将WRO细胞分组并进行处理:1)空白对照组、Cd(5,10,15及30 min)处理组、E2(15 min)处理组和G1(15 min)处理组;2)空白对照组、GPER1特异性拮抗剂(G15)处理组、Cd+G15处理组、E2处理组和E2+G15处理组;3)空白对照组、Cd处理组、Cd+scramble siRNA处理组和Cd+GPER1 siRNA处理组;Western blot检测上述三种情况中各组磷酸化ERK 和AKT及总的ERK 和AKT蛋白表达水平;将WRO细胞分为6组进行处理:空白对照组、Cd处理组、Cd+G15处理组、Cd+ERK 抑制剂(PD98095)处理组、Cd+AKT抑制剂(LY294002)处理组和Cd+GPER1 siRNA处理组,MTT法检测细胞增殖。结果WRO为GPER1阳性;低浓度Cd促进细胞增殖(P<0.05),500 nmol/L Cd效果最明显;随Cd处理时间延长,ERK和AKT蛋白磷酸化水平升高(P<0.05),15min时最高;G15或GPER1siRNA抑制Cd或E2诱导的ERK和AKT蛋白磷酸化水平上升(P<0.05);G15、LY294002、PD98059及GPER1 siRNA减弱Cd诱导的细胞增殖。(P<0.05)。结论镉通过GPER1激活GPER1-ERK/AKT信号通路促进甲状腺癌WRO细胞增殖。

关键词: 镉, GPER1, 甲状腺癌, 增殖

Abstract: ObjectiveTo investigate the proliferative effect of Cadmium on thyroid cancer cell WRO and the involvement of GPER1 in this process.Methods Expression of GPER1 in MCF-7, MDA-MB-231 and WRO cell lineswere detected by Western blot; Three cell lines were divided into 7 groups, respectively: control, Cd (250, 500, 750 and 1000 nmol/L), E2 (10 nmol/L) and G1 (10 nmol/L) groups, Cell proliferationwere measured by MTT method; WRO cell were divided into groups: 1) control, Cd (5, 10, 15 and 30 min), E2 (15 min) and G1 (15 min) groups; 2) control, G15, Cd+G15, E2 and E2+G15 groups; 3) control, Cd, Cd+scramble siRNA and Cd+GPER1 siRNA groups; expression of p-ERK, t-ERK, p-AKT or AKT were detected by Western blot under the above three situations; WRO cell were divided into 6 groups: control, Cd, Cd+G15, Cd+PD98095, Cd+LY294002and Cd+GPER1 siRNA groups, cell proliferation was measured by MTT method. Results Thyroid cancer cell WRO expresses GPER1. Cd promoted cell proliferation in low concentrations (P<0.05), 500nmol/L Cd exhibit the most effective proliferative effect. Cd induced rapid phosphorylation of ERK and AKT with time (P<0.05), which peaked at 15min. Cd-induced ERK and AKT phosphorylationwas inhibited (P<0.05) either by inhibitor for GPER1 or siRNA target GPER1.Cd-induced cell proliferationwas suppressed (P<0.05) byG15,LY294002,PD98059 and GPER1 siRNA. ConclusionsGPER1-ERK/AKT signaling pathway was involved in Cd-induced proliferation of GPER1-positive thyroid cancer cell WRO.

Key words: Cadmium, GPER1, thyroid cancer, proliferation

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