摘要
Abstract
Objective To investigate the effect of microRNA-194 (miR-194) on cell proliferation and apoptosis in melanoma cells and potential underlying mechanism.Methods Real-time fluorescence quantitative PCR (qRT-PCR) was utilized to measure miR-194 in normal primary human skin melanocytes (PIG1) and five kinds of malignant melanoma cells (A375,SK-MEL-1,SK-MEL-2,SK-MEL-5 and SK-MEL-28).A375 melanoma cells were transfected with miR-194 mimics or negative control plasmid via lipofectamine 2000 system.The proliferation capability was measured by MTT assay,and apoptosis rate was determined by flow cytometry.Expressions of Cyclin D1 and cleaved Caspase-3 were detected by Western blot.Results The miR-194 level among A375,SK-MEL-1,SK-MEL-2,SK-MEL-5 and SK-MEL-28 was significantly lower than that in PIG1,and fold of PIG1 was (0.18 ± 0.03),(0.25 ± 0.05),(0.37 ± 0.03),(0.39 ± 0.04),and (0.45 ± 0.03) (P＜ 0.05),respectively.OD values at 490 nm on day 0,1,2,3,4,and 5 post transfection in the miR-194 mimics group vs the negative control group were (0.18 ±0.02) vs (0.19±0.03) (P＞ 0.05),(0.27 ± 0.02) vs (0.29 ±0.03) (P＞ 0.05),(0.42 ±0.08) vs (0.45 ± 0.07) (P＞ 0.05),(0.63 ± 0.09) vs (1.17 ± 0.12) (P＜ 0.05),(1.05 ±0.15) vs (2.15 ± 0.21) (P＜ 0.05) and (1.87 ± 0.23) vs (3.18 ± 0.27) (P＜ 0.05),respectively.The apoptosis rate in the miR-194 mimics group vs the negative control group was 24.2％ vs 9.3％ (P＜ 0.05).Compared with the negative control group,the miR-194 mimics group had (0.36 ± 0.04)-fold change and (3.2 ± 0.28)-fold change in Cyclin D1 and cleaved Caspase-3,respectively (P＜ 0.05).Conclusions There is low expression of miR-194 in melanoma cells.Up-regulation of miR-194 can promote apoptosis rate and inhibit proliferation of melanoma cells by down-regulation of Cyclin D1 protein and up-regulation of cleaved Caspase-3 protein.关键词
micR-194/黑色素瘤/增殖/凋亡Key words
miR-194/melanoma/proliferation/apoptosis分类
医药卫生
