周强;李颖;张恩军;刘琨;

• 1:解放军第454医院骨科

摘要(Abstract)：

[目的]通过转染miRNA-137至人骨肉瘤细胞MG-63,观察细胞内TWIST蛋白含量变化及其对细胞增殖活性的影响以探讨miRNA-137在骨肉瘤中的作用及机制。[方法]Target Scan预测miRNA-137与TWIST结合靶位;PCR扩增TWIST基因3’UTR区并克隆至荧光素酶报告基因表达载体;化学法合成miRNA-137 mimics,miRNA-137 inhibitor及NC序列,与荧光素酶报告基因表达载体共转染293细胞,48 h后检测荧光素酶活性;转染miRNA-137至MG-63细胞,转染后48 h,Real Time-PCR检测转染后细胞内miRNA-137含量,Western-blot检测细胞中TWIST蛋白含量变化;转染后0~72 h,CCK-8检测肿瘤细胞增殖活性。[结果]证实了miRNA-137与其靶基因TWIST的结合位点;miRNA-137 mimics转染MG-63细胞,可明显抑制细胞内TWIST蛋白表达,与对照组比较,差异有统计学意义(P<0.05),而miRNA-137 inhibitor转染细胞后可明显增强胞内TWIST蛋白的表达,与对照组比较差异有统计学意义(P<0.05));miRNA-137 mimics可明显抑制MG-63细胞对数期增殖活性,而miRNA-137 inhibitor则使细胞增殖活性进一步增强,相同时间点与对照相比较,均有差异(P<0.05)。[结论]miRNA-137可以通过抑制其靶基因TWIST表达来抑制MG-63细胞增殖活性。

关键词(KeyWords)： miRNA-137;MG-63;荧光素酶;TWIST;细胞活性

Abstract:

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基金项目(Foundation):

作者(Author): 周强;李颖;张恩军;刘琨;

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参考文献(References)：

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