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hsa-miR-203真核表达载体的构建及对K562细胞增殖与凋亡的影响 被引量：6

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摘要 目的构建hsa-miR-203(人微小非编码RNA-203)的真核表达载体,观察其对人慢性粒细胞白血病细胞株(K562)增殖和凋亡的作用。方法构建hsa-miR-203靶向基因的真核表达载体PmiR-203,用脂质体转染法将PmiR-203转染K562细胞;可溶性噻唑盐-8(WST-8)法检测转染后的K562细胞增殖的情况,流式细胞术检测转染后K562细胞的早期凋亡率,实时荧光定量RT-PCR检测bcr/abl mRNA的表达水平,比色法测定caspase-3、caspase-9的活性。结果本法成功构建了重组真核表达载体PmiR-203,流式细胞术结果表明,K562细胞绿色荧光蛋白(GFP)的表达率52.6%。PmiR-203转染K562细胞48 h后,能明显抑制K562细胞的增殖,并促进细胞的早期凋亡。实时荧光定量RT-PCR结果表明,不同浓度的PmiR-203能显著下调bcr/ablmRNA的表达水平,转染后K562细胞的caspase-3、caspase-9的活性明显增强。结论成功构建了hsa-miR-203的真核表达载体PmiR-203,后者主要通过下调bcr/abl融合基因的表达抑制K562细胞的增殖,并通过活化caspase-3、caspase-9促进K562细胞的早期凋亡。
作者 何金花 黎毓光 谢杏仪 陈顺仪 王莉 邹茂贤 黄国贤
机构地区 广州市番禺区中心医院检验科
出处 《临床检验杂志》 CAS CSCD 北大核心 2012年第8期595-598,共4页 Chinese Journal of Clinical Laboratory Science
基金 广东省科技厅社会发展项目(2011B080702011)
关键词 微小RNA 人微小非编码RNA-203 真核细胞表达载体 K562细胞
分类号 R733.72 [医药卫生—肿瘤]
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参考文献8

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共引文献7

同被引文献52

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引证文献6

二级引证文献15

临床检验杂志
2012年 第8期

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