miR-452-5p靶向SOX7促进食管癌KYSE-150细胞增殖、侵袭及EMT

mi R-452-5p promotes the proliferation, invasion and EMT of esophageal cancer KYSE-150 cells via targeting SOX7

目的:检测miR-452-5p在食管鳞状细胞癌(ESCC)中的表达,并探讨其异常表达对食管癌KYSE-150细胞增殖、侵袭能力和EMT进程的影响及其分子机制。方法:收集2012年3月至2015年12月在河北医科大学第四医院就诊的86名ESCC患者的癌组织样本和对应的癌旁组织,用qPCR法检测miR-452-5p及其他相关基因在ESCC组织和细胞中的表达;向KYSE-150细胞中分别转染miR-452-5p mimic或pcDNA3.1-SOX7构建过表达的细胞株。分析miR-452-5p表达与ESCC病理特征和患者5年OS的关系。用MTS、Tanswell法检测miR-452-5p过表达对食管癌KYSE-150细胞增殖、侵袭能力和EMT进程的影响;用双荧光素酶报告基因实验及TOP/FOP报告基因系统检测miR-452-5p与SRY盒转录因子(SOX7)3’UTR区的结合作用及对Wnt/β-catenin通路活化水平的影响。结果:miR-452-5p在ESCC组织中呈明显高表达(P<0.01),并与ESCC患者的淋巴结转移、TNM分期及5年OS密切相关(均P<0.01)。miR-452-5p过表达明显促进食管癌KYSE-150细胞的增殖、侵袭能力及EMT进程(P<0.05或P<0.01)。SOX7是miR-452-5p的直接靶基因,miR-452-5p通过对SOX7的负向调控影响了Wnt通路活化水平(P<0.05或P<0.01),同时,miR-452-5p表达也受Wnt通路活化水平的影响(P<0.05或P<0.01),其可能为Wnt通路下游靶基因。结论:miR-452-5p通过miR-452-5p/SOX7/Wnt/miR-452-5p正反馈环路提高Wnt/β-catenin通路活化水平,进而促进ESCC KYSE-150细胞的增殖、侵袭能力及EMT进程,miR-452-5p有望成为ESCC患者靶向治疗的潜在靶点及预后评估的新型分子标志物。

Objective:To detect the expression of mi R-452-5p in esophageal squamous cell cancer(ESCC)tissues,and to explore the effect of aberrant miR-452-5p expression on the proliferation,invasion and EMT process of esophageal cancer KYSE-150 cells as well as the possible molecular mechanism.Methods:Cancer tissue samples and corresponding paracancerous tissues were collected from 86 ESCC patients treated at the Fourth Hospital of Hebei Medical University from March 2012 to December 2015.The expression levels of mi R-452-5p and other related genes were detected by q PCR method.KYSE-150 cells were transfected with mi R-452-5p mimic or pc DNA3.1-SOX7 to construct over-expressed cell lines.Analysis of the relationship between mi R-452-5p expression and ESCC pathological features and 5-year OS of patients.MTS and transwell assay were performed to assess the effect of mi R-452-5p over-expression on proliferation,invasion and EMT progress of KYSE-150 cells.The interaction between mi R-452-5p and SRY-box transcription factor 7(SOX7)3’UTR and the activation of Wnt/β-catenin pathway were detected by Dual-luciferase reporter gene assay and TOP/FOP reporter gene system.Results:The expression of mi R-452-5p was significantly up-regulated in ESCC tissues(P<0.01).High expression of mi R-452-5p was correlated with the lymph node metastasis,TNM stage and the 5-year OS of ESCC patients(all P<0.01).Over-expression of miR-452-5p significantly promoted proliferation,invasion ability and the EMT process in KYSE-150 cells(P<0.05 or P<0.01).SOX7 was predicted to be the direct target gene of mi R-452-5p.mi R-452-5p affected the activation level of Wnt pathway through targeted regulation of SOX7(P<0.05or P<0.01).At the same time,the expression of mi R-452-5p was also regulated by the activation level of Wnt pathway,and mi R-452-5p might be a downstream target gene of Wnt pathway(P<0.05 or P<0.01).Conclusion:mi R-452-5p promotes the activation level of Wnt pathway through mi R-452-5p/SOX7/Wnt/mi R-452-5p positive feedback loop,which further promoting the proliferation,invasion and the EMT process of esophageal cancer KYSE-150 cells.mi R-452-5p is expected to be a potential target for molecular therapy and a novel molecular marker for the prognosis evaluation of ESCC patients.