miR-26a对葡萄膜黑色素瘤细胞增殖、凋亡、迁移和侵袭的影响及机制研究

发布时间：2018-04-03 00:03

  本文选题：miR-a　切入点：葡萄膜黑色素瘤　出处：《眼科新进展》2017年07期

【摘要】：目的分析miR26a在葡萄膜黑色素瘤中的表达及对葡萄膜黑色素瘤细胞增殖、凋亡、迁移及侵袭的影响。方法对葡萄膜黑色素瘤细胞系SP6.5、M23及正常葡萄膜上皮细胞系ARPE-19行RT-PCR实验,检测3种细胞系中miR-26a的相对表达差异;将葡萄膜黑色素瘤细胞系SP6.5分成miR-26a模拟物组和NC组,分别转染miR-26a mimics和对照序列scramble,用CCK-8实验和流式细胞术分别检测两组细胞增殖和凋亡;采用细胞划痕实验及Transwell实验分别检测两组细胞的迁移和侵袭能力;采用Western blot检测其下游蛋白组蛋白甲基化转移酶果蝇zeste基因增强子同源物2(enhancer of zeste homolog 2,EZH2)的表达水平。结果葡萄膜黑色素瘤细胞系SP6.5中miR-26a相对表达量为0.250±0.029,细胞系M23中为0.350±0.017,正常葡萄膜上皮细胞系ARPE-19中miR-26a相对表达量为1.0,在细胞系SP6.5及M23中miR-26a的相对表达量均低于正常葡萄膜上皮细胞系ARPE-19中miR-26a的相对表达量(均为P0.001)。miR-26a模拟物组在培养72 h、96 h、120 h OD450值(0.69±0.09、1.23±0.15、2.12±0.23)均显著低于NC组(1.39±0.11、2.35±0.25、3.53±0.27),差异均有统计学意义(均为P0.05)。miR-26a模拟物组细胞凋亡率(15.60±2.30)%高于NC组(5.00±0.70)%(P0.01);miR-26a模拟物组划痕愈合率(23.7±2.1)%低于NC组(68.9±5.1)%(P0.01);侵袭细胞数(45.1±3.9)个低于NC组(115.3±8.9)个(P0.01)。miR-26a模拟物组EZH2蛋白相对表达量(0.39±0.09)低于NC组(1.0;P0.01)。结论 miR-26a在葡萄膜黑色素瘤中低表达,过表达miR-26a显著抑制葡萄膜黑色素瘤细胞增殖,促进凋亡,并抑制迁移和侵袭,其可能是通过下调EZH2的表达来发挥作用的。
[Abstract]:Objective to investigate the expression of miR26a in uveal melanoma and its effect on the proliferation, apoptosis, migration and invasion of uveal melanoma cells.Methods RT-PCR assay was performed on the uveal melanoma cell line SP6.5 M23 and the normal uveal epithelial cell line ARPE-19 to detect the relative expression of miR-26a in the three cell lines, and the SP6.5 cell line was divided into miR-26a mimic group and NC group.The proliferation and apoptosis of miR-26a mimics and control sequence were detected by CCK-8 assay and flow cytometry respectively, and the migration and invasion ability of the two groups were detected by cell scratch test and Transwell assay, respectively.Western blot was used to detect the expression of the enhancer 2(enhancer of zeste homolog 2 (EZH2) in Drosophila melanogaster zeste gene.Results the relative expression of miR-26a was 0.250 卤0.029 in uveal melanoma cell line SP6.5, 0.350 卤0.017 in M23 cell line, and 1.0 in normal uveal epithelial cell line ARPE-19. The relative expression of miR-26a in SP6.5 and M23 cell lines was lower than that in normal staphylococcal cell line.钀勮啘涓婄毊缁嗚優绯籄RPE-19涓璵iR-26a鐨勭浉瀵硅〃杈鹃噺(鍧囦负P0.001).miR-26a妯℃嫙鐗╃粍鍦ㄥ煿鍏，

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