miR-320a抑制宫颈癌细胞迁移、侵袭和上皮间质转化

miR-320a inhibits cervical cancer cell migration,invasion and epithelial-mesenchymal transition

目的 探究miR-320a对宫颈癌细胞迁移、侵袭和上皮间质转化的影响及可能机制。方法 通过OncomiR在线数据库分析宫颈癌组织中miR-320家族miRNA表达;通过Kaplan-Meier Plotter在线数据库分析miR-320家族表达与宫颈癌患者总生存率关系;通过ENCORI在线数据库预测TOP2A mRNA 3′-UTR与miR-320a的潜在结合位点,并通过双荧光素酶报告基因实验验证;将miR-320a模拟物及TOP2A过表达质粒转染宫颈癌细胞,qRT-PCR检测TOP2A mRNA表达,Western blot检测TOP2A蛋白、EMT相关蛋白和PI3K/AKT信号通路相关蛋白表达,Transwell检测细胞迁移和侵袭能力。结果 miR-320a在宫颈癌组织中表达显著下调,其高表达患者总生存率高;miR-320a靶向结合并抑制TOP2A表达;过表达miR-320a抑制宫颈癌细胞迁移和侵袭能力,增加E-cadherin蛋白表达并降低N-cadherin、p-PI3K和p-AKT蛋白表达;过表达TOP2A促进过表达miR-320a的宫颈癌细胞迁移和侵袭能力,降低E-cadherin蛋白表达并增加N-cadherin、p-PI3K和p-AKT蛋白表达。结论 miR-320a过表达抑制宫颈癌细胞迁移、侵袭和EMT,这可能与其靶向抑制TOP2A表达从而抑制PI3K/AKT信号通路激活有关。

ObjectiveTo explore the effect and its mechanism of miR-32Oa on the migration,invasion and epithelial-mesenchymal transition of cervical cancer cells.Methods The expression of miR-320 family miRNA in cervical cancer tissues was analyzed by OncomiR online database.Kaplan-Meier Plotter online database was used to analyze the relationship between miR-320 family expression and overall survival rate of cervical cancer patients.ENCORI online database was used to predict the potential binding sites of TOP2A mRNA 3'-UTR and miR-320a,and verified by double luciferase reporter gene assay.Cervical cancer cells were transfected with miR-320a mimics and TOP2A overexpression plasmid.The mRNA expression of TOP2A was detected by qRT-PCR,and the expressions of TOP2A protein,EMT-related protein and PI3K/AKT signaling pathway were detected by Western blot.Transwell assesses cell migration and invasion.Results The expression of miR-320a was significantly down-regulated in cervical cancer tissues,and the overall survival rate of patients with high expression was high.miR-320a targeted binding and inhibition of TOP2A expression.Overexpressions of miR-320a inhibited the migration and invasion of cervical cancer cells,increased the expressions of E-cadherin protein and decreased the expressions of N-cadherin,P-PI3K and p-AKT protein.Overexpression of TOP2A promoted the migration and invasion ability of cervical cancer cells overexpressing miR-320a,decreased the expression of E-cadherin protein and increased the expressions of N-cadherin,p-PI3K and p-AKT protein.Conclusion Overexpression of miR-320a inhibits the migration,invasion and EMT of cervical cancer cells,which may be related to its targeted inhibition of TOP2A expression and thus inhibiting the activation of PI3K/AKT signaling pathway.