| 胡冉 李惠明 王慧萍 韦芳 王丰 王荣花.Neuronatin对人视网膜色素上皮细胞增殖迁移的影响[J].,2014,14(32):6233-6236 |
| Neuronatin对人视网膜色素上皮细胞增殖迁移的影响 |
| The Effect of Neuronatin on Proliferation and Migration in Human RetinalPigment Epithelial Cells |
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| DOI： |
| 中文关键词: Neuronatin(Nnat) 人视网膜色素上皮细胞 细胞增殖 细胞迁移 |
| 英文关键词: Neuronatin Human retinal pigment epithelial cell Cell proliferation Cell migration |
| 基金项目:上海市自然基金项目（09ZR1425000）；上海市自然基金项目（11ZR1429400） |
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| 中文摘要: |
| 目的：研究胚胎时期表达部位广泛、丰度高，而成年后分化表达的印记基因Neuronatin(Nnat)的两种剪接形式Nnat-alpha和Nnat-beta对人视网膜色素上皮细胞（RPE）增殖、迁移的影响。方法：构建Nnat 琢、茁两种剪接形式的表达质粒，转染RPE 获得表达该基因
的稳定表达细胞株；CCK-8 实验检测稳定表达细胞株的增殖能力，流式细胞仪分析细胞周期，细胞划痕实验检测其迁移能力。结果：成功构建了Nnat-alpha和Nnat-beta表达质粒，并获得了Nnat-alpha和Nnat-alpha基因稳定表达PRE 细胞株。CCK-8实验结果显示cNNAT-alpha
组与对照组相比较，增值率为23.33 %（P<0.05），cNNAT-beta组相较于对照组无显著性差异，细胞周期分析cNNAT-alpha组和cNNAT-beta组细胞在G2-S期的百分率分别为18.60 %、11.11 %，对照组细胞的为9.94 %；相较于对照组，cNNAT-alpha组的细胞迁移能力显著增强，cNNAT-beta组的细胞迁移能力微弱增强。结论：Nnat-alpha对RPE有一定的增殖作用，其影响主要在S期；同时，Nnat-alpha显著促进RPE 细胞的迁移能力。 |
| 英文摘要: |
| Objective:To investigate the effect of two splicing forms of Neuronatin (Nnat-alpha and Nnat-beta), which are expressed
widely and abundantly in the fetal period and differentially expressed mainly in the pituitary gland in the adulthood on the proliferation
and migration in human retinal pigment epithelium (RPE) cellline.Methods:To construct two splicing forms (Nnat alpha and Nnat-beta)expressing plasmids and obtain stable RPE celllines after transfecting RPE cellline with the constructs containing the targeting gene;
proliferation, cell cycle analysis and its ability to migrate were evaluated by CCK-8 assay, flow cytometry and cell wound scratch assay
respectively.Results:Nnat-alpha and Nnat-beta expressing plasmids were successfully constructed, Nnat alpha and Nnat-beta gene stable expressing RPEcelllines were obtained. CCK-8 test results showed that pNNAT-alpha increased cell proliferation by 23.33% (P<0.05) compared with the
control pSNAV transfected cellline. Further analysis through flow cytometry revealed that the percentage of G2S-phase cells in the
control pSNAV transfected cellline was 9.94 %, and in pNNAT-alpha and pNNAT beta stable celllines were 18.60% and 11.11% respectively.
Compared with the control cellline, the migration of pNNAT alpha cellline was enhanced significantly, and the ability of migration of
pNNAT beta was enhanced weakly.Conclusion:NNAT-alpha could enhance the proliferation of RPE cellline significantly and the influence
mainly happened on S period and NNAT-a'phacould enhance the migration of RPE celllines significantly. |
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