miR-137基因转染对人眼葡萄膜黑色素瘤细胞侵袭能力的影响及机制

Effect of miRNA-137 transfection on human uveal melanoma cell line invasion

目的探讨miR-137基因转染对人眼葡萄膜黑色素瘤细胞系M23细胞侵袭能力的影响及其机制。方法将正常培养的M23细胞分为转染组和对照组,每组3孔,分别采用LipofectamineTM2000瞬时转染miR-137 mimic及miR-137 mimics阴性对照。转染18 h后用Transwell侵袭试验检测两组细胞侵袭能力(穿膜细胞数);48 h后采用Western blot法检测细胞基质金属蛋白酶(MMP)-2和MMP-9表达,采用ELISA法检测细胞培养上清液中MMP-2和MMP-9水平。结果转染组穿膜细胞数明显少于对照组(P均<0.05);MMP-2和MMP-9蛋白表达低于对照组(P均<0.05);细胞培养上清液中MMP-2和MMP-9水平低于对照组(P均<0.05)。结论 miR-137基因转染对M23细胞的侵袭能力有抑制作用;其机制可能为抑制细胞MMP-2和MMP-9表达。

Objective To investigate the effects of miR-137 gene transfection on human uveal malignant melanoma cell lines M23.Methods miR-137 mimics were transfected to M23 by Lipofectamine package Lipofectamine TM 2000.Cell invasion was detected by transwell invasion assay 18 h after transfection .MMP-2 and MMP-9 proteins expression were in-vestigated by Western blot and ELISA .Results Transwell invasion assay showed that the number of M 23 cells in miR-137 mimics group was significantly lower than negative control group .Western blot and ELISA confirmed that miR-137 mimics could significantly inhibit the expressions of MMP-2/-9 protein and secreting .Conclusion miR-137 gene transfection can inhibit the invaion of human uveal malignant melanoma cell by reduce the expression of MMP -2 and MMP-9.