大黄素通过TGF-β1/SMAD4通路对子宫内膜癌细胞生物学行为的影响及其机制

Influence and mechanism of Emodin on biological behavior of endometrial carcinoma cells through TGF-β1/SMAD4 pathway

目的:探讨大黄素(Emodin)对子宫内膜癌细胞增殖、迁移、侵袭、上皮间质转化(epithelial mesenchymal transformation,EMT)的影响及作用机制。方法:使用不同浓度的大黄素(0.5~10.0μmol/L)分别处理人子宫内膜癌HEC-1B细胞,检测细胞增殖确定最佳给药浓度。HEC-1B细胞分为对照组(Control组)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)组、Emodin组、Emodin+TGF-β1组、Disitertide(TGF-β1抑制剂)组,分别检测HEC-1B细胞增殖、凋亡、迁移、侵袭及E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、TGF-β1、SMAD家族成员4(mothers against decapentaplegic homolog 4,SMAD4)蛋白表达。建立荷瘤小鼠模型检验大黄素对子宫内膜癌移植瘤生长的影响。结果:0.5~10.0μmol/L的大黄素可显著抑制HEC-1B细胞增殖,选择浓度为2.5μmol/L的大黄素进行后续实验。与Control组比较,TGF-β1组HEC-1B细胞集落形成数、迁移与侵袭数及N-cadherin、Vimentin、TGF-β1表达水平显著升高,E-cadherin、SMAD4表达水平降低(P<0.05),Emodin组与Disitertide组HEC-1B细胞集落形成数、迁移与侵袭数及N-cadherin、Vimentin、TGF-β1表达水平显著降低,细胞凋亡率及E-cadherin、SMAD4表达水平显著升高(P<0.05);与Emodin组比较,Emodin+TGF-β1组HEC-1B细胞集落形成数、迁移与侵袭数及N-cadherin、Vimentin、TGF-β1表达水平显著升高,细胞凋亡率及E-cadherin、SMAD4表达水平降低(P<0.05);大黄素可显著抑制移植瘤生长。结论:大黄素通过调控TGF-β1/SMAD4通路抑制子宫内膜癌肿瘤生长。

Objective:To investigate the influence and mechanism of Emodin on proliferation,migration,invasion and epithelial mesenchymal transformation(EMT)of endometrial carcinoma cells.Methods:Different concentrations of Emodin(0.5~10.0μmol/L)were used to treat human endometrial carcinoma HEC-1B cells respectively,and cell proliferation was detected to determine the optimal drug concentration.HEC-1B cells were divided into Control group,transforming growth factor-β1(TGF-β1)group,Emodin group,Emodin+TGF-β1 group,and Disitertide(TGF-β1 inhibitor)group.The proliferation,apoptosis,migration and invasion of HEC-1B cells,and the expression of E-cadherin,N-cadherin,Vimentin,TGF-β1,mothers against decapentaplegic homolog 4(SMAD4)proteins were detected respectively.A tumor bearing mouse model was established to test the effect of Emodin on the growth of endometrial carcinoma xenografts.Results:Emodin of 0.5~10.0μmol/L could obviously inhibit the proliferation of HEC-1B cells.Emodin of 2.5μmol/L concentration was selected for subsequent experiments.Compared with the Control group,the number of HEC-1B cell colonies,migration and invasion,and the expression levels of N-cadherin,Vimentin,TGF-β1 in TGF-β1 group increased obviously,the expression levels of E-cadherin and SMAD4 decreased(P<0.05),the number of HEC-1B cell colonies,migration and invasion,and the expression levels of N-cadherin,Vimentin and TGF-β1 reduced obviously in Emodin group and Disitertide group,the apoptosis rate and the expression levels of E-cadherin and SMAD4 increased obviously(P<0.05).Compared with Emodin group,the number of HEC-1B cell colonies,migration and invasion,and the expression levels of N-cadherin,Vimentin and TGF-β1 increased obviously in Emodin+TGF-β1 group,the apoptosis rate and the expression levels of E-cadherin and SMAD4 reduced obviously(P<0.05).Emodin was able to obviously inhibit the growth of transplanted tumor.Conclusion:Emodin inhibits the growth of endometrial carcinoma by regulating TGF-β1/SMAD4 pathway.