miR-185靶向Six1对肺癌上皮-间质转化过程的影响及其机制

摘要

Purpose To study the molecular mechanism of miR-185 affecting the migration and invasion of human lung cancer cell.Methods MiR-185 overexpression was obtained by transfection of miR-185 mimic in lung squamous cell carcinoma cell line H520 and A549,transwell assay and cell scratch assay were used to detection of cell migration and invasion.The luciferase reporter assay confirmed that miR-185 targets the Six1 gene.qRT-PCR and Western blot were used to detect the impact of miR-185 cells Six1 gene expression.Western blot was used to detect the effect of miR-185 overexpression on the epithelialmesenchymal transition of lung cancer cells.Results miR-185overexpression reduced migration and invasion of lung cancer cells (P ＜ 0.05),increased epithelial cell marker E-cadherin expression (P ＜ 0.01),and decreased the expression of mesenchymal cell markers vimentin of (P ＜ 0.01).After overexpression of miR-185 in H520 cells,the expression level of Six1gene was reduced (P＜0.01).MiR-185 regulated the migration and invasion of lung cancer cells by targeting the Six1 gene.Conclusion MiR-185 targets the Six1 gene to regulate the EMT pathway of human lung cancer cells.%目的 探讨miR-185影响人肺癌细胞迁移和侵袭能力的分子机制.方法 在肺癌细胞系H520和A549中转染miR-185mimic获得miR-185过表达,利用Transwell实验和细胞划痕实验检测细胞的迁移和侵袭能力;荧光素酶报告实验验证miR-185靶向Six1基因;采用qRT-PCR和Western blot法检测miR-185对细胞中Six1基因表达水平的影响;Western blot法检测miR-185过表达对肺癌细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)过程的影响.结果 miR-185过表达降低肺癌细胞的迁移和侵袭能力(P＜0.05),上皮细胞标志物E-cadherin表达升高(P＜0.01),间质细胞标志物vimentin表达降低(P＜0.01),H520细胞过表达miR-185后,Six1基因表达水平降低(P＜0.01),miR-185调控肺癌细胞的迁移和侵袭是通过靶向Six1基因实现的.结论 miR-185靶向Six1基因调控人肺癌细胞EMT途径.