[1]李娜,曹娟,郁继国,等.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[J].眼科新进展,2017,37(7):619-622.[doi:10.13389/j.cnki.rao.2017.0157]
　LI Na,CAO Juan,YU Ji-Guo,et al.Effects of miR-26a on proliferation，apoptosis，migration and invasion of uveal melanoma cell and its mechanism[J].Recent Advances in Ophthalmology,2017,37(7):619-622.[doi:10.13389/j.cnki.rao.2017.0157]
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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/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
37卷
期数:
2017年7期
页码:
619-622
栏目:
实验研究
出版日期:
2017-07-05

文章信息/Info

Title:
Effects of miR-26a on proliferation，apoptosis，migration and invasion of uveal melanoma cell and its mechanism
作者:
李娜; 曹娟; 郁继国; 李亚兰
430000　湖北省武汉市，华中科技大学同济医学院附属武汉中心医院眼科
Author(s):
LI Na; CAO Juan; YU Ji-Guo; LI Ya-Lan
Department of Ophthalmology，the Central Hospital of Wuhan，Tongji Medical College，Huazhong University of Science and Technology，Wuhan 430000，Hubei Province，China
关键词:
miR-26a; 葡萄膜黑色素瘤; 增殖; 凋亡; 侵袭; 转移; EZH2
Keywords:
miR-26a; uveal melanoma; proliferation; apoptosis; invasion; migration; EZH2
分类号:
R773.9
DOI:
10.13389/j.cnki.rao.2017.0157
文献标志码:
A
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P<0.001）。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P<0.05）。miR-26a模拟物组细胞凋亡率（15.60±2.30）%高于NC组（5.00±0.70）%（P<0.01）；miR-26a模拟物组划痕愈合率（23.7±2.1）%低于NC组（68.9±5.1）%（P<0.01）；侵袭细胞数（45.1±3.9）个低于NC组（115.3±8.9）个（P<0.01）。miR-26a模拟物组EZH2蛋白相对表达量（0.39±0.09）低于NC组（1.0；P<0.01）。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 analyze the effects of miR-26a on proliferation，apoptosis，migration and invasion of uveal melanoma cell.Methods　The expressions level of miR-26a in the uveal melanoma cell lines SP6.5 and M23 and normal cell line ARPE-19 was detected by qRT-PCR.The SP6.5 cell line was divided into miR-26a mimics group （transfect with miR-26a mimics） and NC group （transfect with scramble by Lipofectamine 2000）.The proliferation and apoptosis ability were measured by CCK-8 assay and flow cytometry.The cell wound scratch assay and transwell assay were used to detect the migration and invasion ability.The expression level of enhancer of zeste homolog 2 （EZH2） protein was measured by Western blot.Results　The expressions level of miR-26a in uveal melanoma cell line SP6.5 was 0.250±0.029，M23 was 0.350±0.017，which was significantly lower than 1.0 in normal cell line ARPE-19 （all P<0.001）.The OD450 value at 72 hours，96 hours and 120 hours in miR-26a mimics group （0.69±0.09，1.23±0.15，2.12±0.23） were significantly lower than those in NC group （1.39±0.11，2.35±0.25，3.53±0.27） （all P<0.05）.The apoptosis rate of miR-26a mimics group （15.60%±2.30%） was significantly higher than that of NC group （5.00%±0.70%） （P<0.01）.The wound healing rate of miR-26a mimics group （23.7%±2.1%） was significantly lower than that of NC group （68.9±5.1%）（P<0.01）.While the invasive cell number （45.1±3.9） was also significantly less than NC group （115.3±8.9）（P<0.01）.The expression level of EZH2 protein in miR-26a mimics group （0.39±0.09） was significantly lower than that of NC group （1.0，P<0.01）.Conclusion　MiR-26a is low expressed in uveal melanoma cells.Over expression of miR-26a inhibit the proliferation，migration and invasion of uveal melanoma cell，and promote the apoptosis，which may be associated with down-regulated expression of EZH2.

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更新日期/Last Update: 2017-08-02