miR-205对黑色素瘤A375细胞增殖、黏附和迁移的影响及机制探讨

摘要

目的 探讨miR-205对人皮肤恶性黑色素瘤细胞系A375增殖、黏附和迁移的影响及其可能机制.方法 将miR-205 mimics用阳离子脂质体LipofectamineTM2000转染A375细胞,CCK-8法测细胞增殖变化；细胞黏附实验和划痕实验检测细胞黏附和迁移能力变化.Western blot检测PTEN、总AKT和磷酸化AKT蛋白表达变化.结果 上调miR-205表达后实验组的细胞增殖、黏附和迁移能力均比空白对照组和阴性对照组减低(P均＜0.05)；Western blot结果显示上调miR-205表达能增加PTEN蛋白表达,减低磷酸化AKT蛋白水平,但对非AKT蛋白表达无影响.结论 miR-205可能通过调控PTEN/AKT通路抑制黑色素瘤细胞的增殖、黏附和迁移能力.%Objective To investigate the effects of miR-205 on proliferation, adhesion and migration in human skin malignant melanoma cell line A375 and further elucidate its mechanisms. Methods miR-205 mimics was transfected A375 by lipofectamine package. Cell proliferation was studied by Cell Counting Kit-8 ( CCK-8 ) assay. The wound migration assay and adhesion assay were utilized to analyze the migration and adhesion of A375 cells in vitro after miR-205 over-expression. The expressions of proteins were investigated by Western blot. Results miR-205 significantly inhibited cell proliferation, adhesion and migration. PTEN protein was markedly increased and the phosphorylation of AKT was markedly reduced by miR-205 mimics treatment, respectively. But miR-205 had no effect on unphosphorylated AKT. Conclusion miR-205 exhibits inhibitory effects on proliferation, adhesion and migration in human skin malignant melanoma cell line, and its action mechanisms may involve in PTEN/AKT pathway.