摘要
目的探讨microRNA-7(miR-7)对肝癌细胞株MHCC-97H上皮-间质转化(EMT)的影响及作用机制。方法将构建的miR-7真核载体单独及分别联合野生型或突变型EGFR 3’-UTR真核载体共转染MHCC-97H细胞,采用Realtime PCR、Western blot检测EMT标识蛋白E-cadherin、β-catenin、N-cadherin和Vimentin的表达;细胞划痕、Transwell实验检测细胞侵袭和迁移能力的变化;双荧光素酶实验分析miR-7对EGFR的调控作用。结果与正常组相比,转染miR-7真核载体组细胞E-cadherin、β-catenin表达均显著升高(P<0.01),N-cadherin、Vimentin表达则明显降低(P<0.01);同时,细胞的侵袭、迁移能力均明显减弱(P<0.01);分别与转染GV272+EGFR 3’-UTR野生型和转染GV272/miR-7+EGFR 3’-UTR突变型相比,转染GV272/miR-7+EGFR 3’-UTR野生型组萤火虫荧光素酶活性显著降低(P<0.01)。结论 miR-7可通过与EGFR 3’-UTR结合而抑制EGFR信号通路,降低MHCC-97H细胞侵袭、迁移能力,进而抑制MHCC-97H细胞EMT。
Objective To investigate the effect and mechanism of microRNA-7 （miR-7） on epithelial-mesenchymal transition of MHCC-97H cell. Methods MiR-7 eukaryotic vector was co-transfected into wild-type or mutant EG- FR 3＇ -UTR eukaryotic vector, respectively. The expression of E-cadherin, β-catenin, N-cadherin and Vimentin were detected by real-time PCR and Western blot. Meanwhile, cell scratch test and transwell assay was used to de- tect the invasion and migration ability of cells. Dual luciferase assay was used to analyze the effect of miR-7 on EG- FR of 3＇ -UTR. Results Compared with normal group, the expression of E-cadherin and β-catenin was significantly increased in GV268/miR-7 group （P 〈0. 01 ）, but the expression of N-cadherin and Vimentin was significantly decreased （P 〈0. 01 ）. Meanwhile ,the invasion and migration ability was also significantly decreased （P 〈0. 01 ）. Compared with EGFR 3＇ -UTR wild-type group and GV272/miR-7 ＋ EGFR 3 ＇ -UTR mutation group respectively, the luciferase activity of GV272/miR-7 ＋ EGFR 3＇ -UTR wild-type group was significantly higher （ P 〈0. 01 ）. Conclusion miR-7 could inhibit the EGFR signaling pathway by binding to EGFR 3＇ -UTR, decrease the invasion and migration of MHCC-97H cells and inhibit the EMT of MHCC-97H cells.
出处
《安徽医科大学学报》
CAS
北大核心
2018年第1期29-34,共6页
Acta Universitatis Medicinalis Anhui
基金
安徽省自然科学基金(编号:1608085MH198)
