miR-16在葡萄膜黑色素瘤细胞中的表达变化及其对细胞增殖的影响

发布时间：2018-11-26 16:30

【摘要】：目的观察miR-16在葡萄膜黑色素瘤细胞中的表达变化,及其对细胞增殖的影响。方法培养葡萄膜黑色素瘤细胞系SP6.5及正常葡萄膜黑色素细胞系UM-U-95。采用实时定量PCR法检测SP6.5细胞、UM-U-95细胞中的miR-16。将SP6.5细胞分为miR-16组、对照组和miR-16抑制剂组。其中miR-16组转染miR-16 mimics,对照组转染scramble(阴性对照序列),miR-16抑制剂组转染miR-16抑制剂。分别于转染24、48、72 h采用MTT法测算各组相对活细胞百分比;转染72 h后采用克隆形成实验测算克隆形成率;转染72h后,采用Western blotting法检测各组细胞中的成纤维细胞生长因子2(FGF2)蛋白。结果 SP6.5细胞、UM-U-95细胞中miR-16相对表达量分别为0.22±0.02、1.0,SP6.5细胞中miR-16的相对表达量低于UM-U-95细胞(P0.01)。转染后48、72 h miR-16组相对活细胞百分比低于对照组,miR-16抑制剂组相对活细胞百分比高于对照组(P均0.05)。转染72 h后,miR-16组、对照组、miR-16抑制剂组克隆形成率分别为20.15%±2.53%、42.63%±4.42%、68.36%±6.25%,miR-16组克隆形成率低于对照组,miR-16抑制剂组克隆形成率高于对照组(P均0.05)。miR-16组、对照组、miR-16抑制剂组细胞中FGF2蛋白相对表达量分别为0.27±0.02、1.0、5.64±0.58,miR-16组FGF2蛋白相对表达量低于对照组,miR-16抑制剂组FGF2蛋白相对表达量高于对照组(P均0.05)。结论 miR-16在葡萄膜黑色素瘤细胞中表达下调。miR-16过表达可抑制葡萄膜黑色素瘤细胞增殖、降低克隆形成率,抑制miR-16表达可增强葡萄膜黑色素瘤细胞的增殖能力。miR-16对葡萄膜黑色素瘤细胞增殖能力的影响可能与调节FGF2蛋白表达有关。
[Abstract]:Objective to observe the expression of miR-16 in uveal melanoma cells and its effect on cell proliferation. Methods cultured uveal melanoma cell line SP6.5 and normal uveal melanoma cell line UM-U-95. Detection of miR-16. in SP6.5 cells and UM-U-95 cells by Real-time quantitative PCR SP6.5 cells were divided into miR-16 group, control group and miR-16 inhibitor group. MiR-16 group transfected miR-16 mimics, control group with scramble (negative control sequence), miR-16 inhibitor group transfected miR-16 inhibitor. The relative percentage of living cells in each group was calculated by MTT method at 72 h after transfection, and the colony formation rate was calculated by clone formation experiment 72 h after transfection. After 72 hours of transfection, fibroblast growth factor 2 (FGF2) protein was detected by Western blotting assay. Results the relative expression of miR-16 in SP6.5 cells and UM-U-95 cells was 0.22 卤0.02 ~ 1.0P ~ (6.5), respectively, which was lower than that in UM-U-95 cells (P0.01). The percentage of relative living cells in miR-16 group was lower than that in control group at 48 ~ 72 h after transfection, and the relative live cell percentage in miR-16 inhibitor group was higher than that in control group (P < 0. 05). After 72 hours of transfection, the clone formation rate of miR-16 group, control group and miR-16 inhibitor group was 20.15% 卤2.53% 卤42.63% 卤4.42% respectively. The clone formation rate of miR-16 group was lower than that of control group (68.36% 卤6.25%). The relative expression of FGF2 protein in miR-16 group, miR-16 inhibitor group and miR-16 inhibitor group was 0.27 卤0.021.0 卤5.64 卤0.58, respectively. The relative expression of FGF2 protein in miR-16 group was lower than that in control group, and the relative expression of FGF2 protein in miR-16 inhibitor group was higher than that in control group (P 0.05). Conclusion the expression of miR-16 is down-regulated in uveal melanoma cells, and the overexpression of miR-16 can inhibit the proliferation of uveal melanoma cells and decrease the clone formation rate. Inhibiting the expression of miR-16 could enhance the proliferative ability of uveal melanoma cells. The effect of miR-16 on the proliferation of uveal melanoma cells may be related to the regulation of FGF2 protein expression.
【作者单位】：武汉大学中南医院;
【分类号】：R739.7

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