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CDK2干扰RNA新靶点的发现及对人脑胶质瘤生长的影响

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摘要 目的:构建4个新靶点的人CDK2干扰RNA真核表达载体,转染人脑胶质细胞瘤SHG44细胞,经RT-PCR检测mRNA的表达MTT法检测细胞增殖,获得干扰效果最好的真核表达载体,为继续探讨CDK2在SHG44细胞中的作用提供有价值的资料及实验基础。方法:1.构建4个新靶点CDK2干扰RNA真核表达载体并用双酶切和测序鉴定。2.脂质体法瞬时转染上述4个载体到SHG44细胞株。3.通过RT-PCR比较转染后CDK2mRNA的表达量,MTT法检测细胞增殖。结果:1.成功构建了4个新靶点的CDK2干扰RNA真核表达载体PGPU6/GFP/Neo-CDK2-1、PGPU6/GFP/Neo-CDK2-2、PGPU6/GFP/Neo-CDK2-3、PGPU6/GFP/Neo-CDK2-4。2.CDK2 mRNA表达和细胞增殖明显受到抑制,并获得了效果最好的CDK2干扰RNA真核表达载体。结论:.成功构建并筛选出了效果最好的新靶点CDK2干扰RNA真核表达载体,使SHG44细胞的CDK2 mRNA的表达明显降低。
出处 《内蒙古医学院学报》 2012年第S7期915-917,920,共4页 Acta Academiae Medicinae Neimongol
基金 内蒙古自治区自然基金(2012MS1147)
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