缺氧及缺氧相关miR-210调节胃癌细胞MGC803的上皮间充质转化

摘要

Objective To explore the effect of anoxic microenvironment and hypoxia related microRNA-210 (miR-210) on the epithelial-mesenchymal transition (EMT) in gastric cancer cell line 803.Methods The mRNA relative expressions of E-cadherin and Twist1 were detected by qRT-PCR in gastric cancer cell line after being induced hypoxia by CoCl2 (100,200 and 300 μ mol/L),which was compared with normoxic group (0 mol/L).The cell proliferation of gastric cancer cell line was observed by cell proliferation assay.The relative expressions of E-cadherin and Twist 1 were detected in gastric cancer cell line after transfecting miR-210 mimics.Results With the increase of the concentrations of CoCl2,the relative expression of miR-210 and the proliferation rate of gastric cancer cells showed a tendency of increase first and then decrease (P ＜ 0.05).The relative expression of E-cadherin mRNA was lower in hypoxia group than that of normoxic group.The relative expression of Twist1 mRNA was higher in hypoxia group than that of normoxic group.The relative expressions of Ecadherin mRNA were significantly higher in 200 and 300 μ mol/L CoCl2 groups than that of 100 μmol/L CoCl2 group (P ＜0.05).There was no significant difference in the relative expression of Twist1 mRNA between the three CoCl2 groups.The relative expression of Twist1 mRNA was higher in the transfection group than that of control group.The relative expression of E-cadherin mRNA was significantly lower in the transfection group than that of control group (P ＜ 0.05).Conclusion Hypoxia can promote the proliferation and EMT of gastric cancer cells.The up-regulation of miR-210 can also promote the procession of EMT,which may be the intermediate link in the process of EMT induced by hypoxia.%目的 探讨缺氧微环境及缺氧相关微小RNA-210(miR-210)对胃癌细胞MGC803的上皮间充质转化(EMT)过程的影响.方法 不同浓度CoCl2溶液(100、200、300 μmol/L)诱导胃癌细胞缺氧后,qRT-PCR检测缺氧组中EMT标志物E-钙黏蛋白(E-cadherin)和Twist1的mRNA相对表达量,并与常氧组(0μmol/L)进行比较;细胞增殖实验观察不同缺氧程度下胃癌细胞的增殖情况;通过向胃癌细胞转染miR-210 mimics提高miR-210的表达后,qRT-PCR检测转染组与对照组(未转染mimics) E-cadherin及Twist1的mRNA相对表达量.结果 随着CoCl2溶液浓度的增高,miR-210相对表达量、胃癌细胞的增殖率均呈先增后减的趋势(P＜0.05);缺氧组E-cadherin mRNA相对表达量均低于常氧组,Twist1 mRNA相对表达量均高于常氧组;200、300 μmol/L CoCl2组E-cadherin mRNA相对表达量均高于100 μmol/L CoCl2组(P＜0.05),3组Twist1 mRNA相对表达量差异无统计学意义.转染组Twist1 mRNA相对表达量高于对照组,E-cadherin mRNA相对表达量低于对照组(P＜0.05).结论 缺氧可促进胃癌细胞的增殖及EMT过程,miR-210的上调可促进EMT过程,miR-210可能是缺氧诱导EMT过程的中间环节之一.