miRNA-675-5p与食管鳞癌上皮间质转化的相关性研究

Realationship study on miRNA-675-5p and epithelial-mesenchymal transition in esop-hageal squamous cell carcinoma

目的探讨miRNA-675-5p表达水平与食管鳞癌(ESCC)上皮间质转化(EMT)的相关性。方法采用实时定量PCR技术检测miRNA-675-5p在ESCC组织及配对癌旁正常组织中的表达差异,并分析其表达水平与患者临床分期的相关性。脂质体转染ECA109细胞,分为miR-675-recursor组、miR-675-5p-inhibition组和NC组,上调及抑制miRNA-675-5p表达水平,分析改变目的基因表达后对ECA109细胞株迁移侵袭能力的影响,蛋白印迹实验检测干扰表达每组细胞中EMT相关蛋白E-cadherin、N-cadherin、Vimentin表达水平,评估其对ECA109株EMT过程的影响。结果与癌旁正常组织比较,miRNA-675-5p在ESCC组织中表达显著升高(P<0.05),不同分期患者miRNA-675-5p表达比较,差异有统计学意义(P<0.05)。与NC组比较,miR-675-recursor组细胞迁移侵袭能力明显增强,miR-675-5p-inhibition组细胞迁移侵袭能力受到显著抑制(P<0.05)。此外,与NC组比较,miR-675-recursor组E-cadherin蛋白表达降低,N-cadherin、Vimentin蛋白表达升高;miR-675-5p-inhibition组E-cadherin蛋白表达升高,N-cadherin、Vimentin蛋白表达降低(P<0.05)。结论miRNA-675-5p表达与ESCC患者TNM分期相关,miR-675-5p能促进ESCC细胞间质转化从而增强侵袭能力;下调表达能逆转ESCC细胞EMT过程,降低迁移侵袭能力。

Objective To investigated the correlation between the expression of miRNA-675-5p and epithelial-mesenchymal transition(EMT)in esophageal squamous cell carcinoma(ESCC).Methods Real-time quantitative PCR was used to detect the differential expression of miRNA-675-5p in ESCC and normal adjacent tissues,while the correlation between its expression level and clinical stage of patients was analyzed.Liposomes was transfected into ECA109 cell,and divided into miR-675-recursor group,miR-675-5p-inhibition group and NC group.The expression of miRNA-675-5p was up-regulated and inhibited,while the effect of changing the expression of the target gene on the migration and invasion of ECA109 cell lines was analyzed.The expression levels of E-cadherin,N-cadherin,and Vimentin in EMT-associated proteins in each group of cells were detected by western blot experiments,while the effect on the EMT process of ECA109 strain was evaluated.Results Compared with normal adjacent tissues,miRNA-675-5p expression was significantly increased in ESCC tissues(P<0.05),and comparison of miRNA-675-5p expression in patients with different stages,the differences were statistically significant(P<0.05).Compared with the NC group,the miR-675-recursor group had significantly enhanced cell migration and invasion ability,and the miR-675-5p-inhibition group had significantly reduced cell migration and invasion ability(P<0.05).In addition,compared with the NC group,the expression of E-cadherin protein in the miR-675-recursor group was reduced,while the expression of N-cadherin and Vimentin proteins were in creased;the expression of E-cadherin protein in the miR-675-5p-inhibition group was increased,while the expression of N-cadherin and Vimentin proteins were reduced(P<0.05).Conclusion The expression of miRNA-675-5p is related to the TNM stage of ESCC patients.miR-675-5p can promote the interstitial transformation of ESCC cells and thus enhance the invasion ability;down-regulated expression can reverse the EMT process of ESCC cells and reduce the migration and invasion ability.