姜黄素抑制肠道肿瘤上皮间质转化的研究

Curcumin inhibits the epithelial-mesenchymal transition of intestinal cancer

目的:通过姜黄素对肠道肿瘤细胞miR-200a影响,明确其抑制肿瘤细胞EMT机理,揭示其抗肠道肿瘤侵袭转移机制。方法:以人结肠腺癌细胞HCT116为模型,采用PCR技术、基因重组技术、侵袭实验等一系列实验方法研究姜黄素对肿瘤细胞以下方面的影响:miR-200a表达水平,β-连环蛋白、波形蛋白、ZEB1、E-钙粘蛋白mRNA,细胞生长转化、侵袭转移。结果:与空白对照组比较,终浓度为3.68mg/L、7.36mg/L和14.73mg/L的姜黄素和3.00mg/L的顺铂可以显著抑制HCT116细胞软琼脂集落形成、HCT116细胞侵袭、HCT116细胞β-连环蛋白mRNA和波形蛋白mRNA表达、HCT116细胞ZEB1蛋白翻译,可以显著促进HCT116细胞microRNA-200a表达、促进E-钙粘蛋白蛋白翻译,姜黄素效应均表现出浓度依赖性。结论:姜黄素可以抑制肠道肿瘤细胞上皮间质转化的表型,即抑制HCT116软琼脂集落形成和细胞侵袭。其机制在于提高HCT116细胞microRNA-200a水平,抑制细胞β-连环蛋白mRNA、波形蛋白mRNA转录,抑制细胞ZEB1蛋白翻译、促进E-钙粘蛋白蛋白翻译。

Objective： By investigating the effect of curcumin on miR-200 a expression of intestinal cancer cells,to elucidate the mechanism that curcumin inhibits epithelial-mesenchymal transition（ EMT）,invasion and metastasis of intestinal cancer. Methods： Soft agar assay for colony formation,cell invasion assay,real-time PCR and western blot analyses were carried out to investigate the effect of curcumin on colony formation,cell invasion,miR-200 a expression,mRNA levels of β-catenin and vimentin,protein levels of ZEB1 and E-cadherin of HCT116 cells,respectively. Results： Compared with the control group,curcumin at concentrations of 3. 68mg/L,7. 36mg/L and 14. 73mg/L,cisplatin at a concentration of 3. 00mg/L all significantly suppressed colony formation,cell invasion,mRNA levels of β-catenin and vimentin,ZEB1 protein level of HCT116 cells,but heightened miR-200 a expression and E-cadherin protein level. In addition,no difference in potencies was observed between curcumin at 14. 73mg/L and cisplatin at 3. 00mg/L. Conclusions： Curcumin may inhibit the phynotype of intestinal cancer EMT,HCT116 cell colony formation and invasion,by increasing miR-200 a level,reducing mRNA levels of β-catenin and vimentin,and ZEB1 protein expression,also enhancing E-cadherin protein expression.