摘要
目的探讨miR-489在结肠癌干细胞表型和上皮间质转换中的作用和调控机制。方法 RT-q PCR检测多个人结肠癌细胞系(HT29、SW480、SW620和HCT116)和正常肠道上皮细胞HIEC中miR-489表达;采用基因转染技术提高结肠癌细胞中miR-489表达,Western blot检测过表达miR-489对结肠癌干细胞标志物CD133、CD44、Ep CAM、ALDH1和上皮细胞标志物E-cadherin、间质细胞标志物vimentin和N-cadherin的表达;RT-q PCR和Western blot检测TWIST1 mRNA和蛋白表达水平。结果 4个结肠癌细胞系中的miR-489相对表达量显著低于正常肠道上皮细胞HIEC中的表达(P<0.05)。在HT29和HCT116细胞中过表达miR-489,4个结肠癌干细胞标志物表达降低(P<0.05);上皮细胞标志物表达增加(P<0.05);间质细胞标志物表达减少(P<0.05)。TWIST1是miR-489潜在靶基因,在2个细胞系中过表达miR-489,TWIST1 mRNA和蛋白表达水平均显著降低(P<0.05)。结论 miR-489在结肠癌细胞中低表达,过表达miR-489可能通过调控TWIST1抑制结肠癌干细胞表型和上皮间质转换。
Objective To explore the effects and regulation mechanism of miR-489 on the stem cell phenotype and epithelial-mesenchymal transition of colon cancer cells in vitro. Methods The miR-489 expression of colon cancer cell lines（ HT29,SW480 and SW620,HCT116） and normal intestinal epithelial cells HIEC were detected by RT-q PCR,the miR-489 expression in colon cancer cells was raised by gene transfection technology,the colon cancer stem cell markers CD133,CD44,Ep CAM,ALDH1 and epithelial marker E-cadherin,mesenchymal cells markers vimentin and N-cadherin were detected by Western blot. The expression of TWIST1 mRNA and protein were detected by RT-q PCR and Western blot. Results The miR-489 relative expression in four kinds of colon cancer cells（ HT29,SW480 and SW620,HCT116） were significantly lower than that of normal HIEC intestinal epithelial cells,the difference was statistically significant（ P 0. 05）. Up-regulation of miR-489 expression in HT29 and HCT116 cells leaded to lower expression of colon cancer stem cell marker CD133,CD44,Ep CAM ALDH1 and mesenchymal cells markers Vimentin,N-cadherin,higher expression of epithelial markersE-cadherin（ P〈0. 05）. Also,up-regulation of miR-489 expression in HT29 and HCT116 cells leaded to lower expression of TWIST1 mRNA and protein（ P〈0. 05）. Conclusions miR-489 is down-regulated in colon cancer cells and up-regulation of miR-489 expression inhibits stem cell phenotype and epithelial-mesenchymal transition through targeting TWIST1 in colon cancer.
出处
《基础医学与临床》
CSCD
2018年第1期51-56,共6页
Basic and Clinical Medicine
基金
陕西省社会发展科技攻关项目(2014SF2-16)
