MicroRNA-205抑制黑素瘤细胞迁移和侵袭的作用及其机制

发布时间：2018-07-31 17:03

【摘要】：目的探讨microRNA-205(miR-205)表达对人类黑素瘤细胞株(A375和A2058)迁移和侵袭的作用及其分子机制。方法转染miR-205过表达慢病毒(Lenti-miR-205)构建miR-205过表达的黑素瘤细胞A375和A2058细胞系(miR-205组),转染空白对照慢病毒作为对照细胞系(NC组)。通过划痕实验和Transwell实验分别检测两组细胞迁移和侵袭能力,激光共聚焦扫描显微镜观察细胞形态和表达钙黏蛋白E(E-cadherin)的细胞数量与荧光强度,蛋白质印迹法检测E-cadherin、Twist、整合素(intergrin)、波形蛋白(vimentin)、Zeb1、基质金属蛋白酶(MMP)-2、MMP-9蛋白表达。结果培养8 h和24 h时miR-205组A375与A2058细胞的迁移能力均大于NC组(P0.01);miR-205组两种细胞的侵袭能力均低于NC组(P0.01)。MiR-205过表达后A375和A2058细胞均由梭形、基质型向圆形、表皮型转化。与NC组相比,miR-205组细胞E-cadherin蛋白表达增加(P0.01),且表达E-cadherin的细胞数量和荧光强度均增加;Twist、intergrin、vimentin、Zeb1、MMP-2和MMP-9蛋白表达水平降低(P0.01)。结论 MiR-205通过诱导E-cadherin的表达逆转上皮间质转化,从而抑制黑素瘤细胞的迁移和侵袭。
[Abstract]:Objective to investigate the effect of microRNA-205 (miR-205) expression on migration and invasion of human melanoma cell lines (A375 and A2058) and its molecular mechanism. Methods miR-205 overexpression lentivirus (Lenti-miR-205) was transfected into A375 and A2058 melanoma cells with miR-205 overexpression (miR-205 group), and blank control lentivirus was transfected as control cell line (NC group). The migration and invasion ability of the two groups were detected by scratch test and Transwell assay. The cell number and fluorescence intensity of the cells expressing cadherin E (E-cadherin) and morphology were observed by confocal laser scanning microscopy. The expression of E-cadherin Twist, integrin (intergrin), vimentin (vimentin) Zeb1, matrix metalloproteinase (MMP)-2 and MMP-9 were detected by Western blot. Results the migration ability of A375 and A2058 cells in miR-205 group was higher than that in NC group (P0.01) at 8 h and 24 h. The invasion ability of A375 and A2058 cells in miR-205 group was lower than that in NC group (P0.01). MiR-205 overexpression was lower than that in NC group (P0.01). Compared with NC group, the expression of E-cadherin protein was increased (P0.01), and the number and fluorescence intensity of E-cadherin were increased (P0.01). The expression of MMP-2 and MMP-9 protein were decreased (P0.01). Conclusion MiR-205 inhibits the migration and invasion of melanoma cells by inducing E-cadherin expression to reverse epithelial mesenchymal transformation.
【作者单位】：复旦大学附属华山医院皮肤科;
【基金】：上海市卫生局科研课题(20134363)~~
【分类号】：R739.5

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