摘要
目的探讨RNA干扰技术抑制Midkine对人乳腺癌癌细胞株MCF-7细胞凋亡的影响。方法人工合成抑制Midkine基因的siRNA片段,通过脂质体转染到MCF-7细胞内,应用RT-PCR检测bcl-2 mRNA表达,流式细胞术检测细胞线粒体膜电位变化,通过测定光密度值检测Caspase-3活性。结果转染siRNA后的MCF-7细胞,bcl-2 mRNA表达显著下降、细胞线粒体膜电位明显下降和Caspase-3酶活性升高。结论MK-siRNA通过下调bcl-2基因,而改变细胞线粒体膜电位使Caspase-3酶活性升高诱导细胞的凋亡。
Objective To explore the apoptosis of MCF-7 cells induced by siRNA-mediated midkine silencing. Methods The selected siRNA targeting Midkine （MK） was transfected into MCF-7 cells by LipofectaminTM2000. The expression of bcl-2 mRNA was detected by RT-PCR.The mitochondrial membrane potential was determined by flow cytmeter analysis and the activity of caspase-3 was examined by colorirnctric. Results For the MCF-7 cell transfected with siRNA, the expression of Bcl-2 mRNA was significantly decreased. The mitechondrial membrane potential was loss and the activity of caspase-3 was increased. Contusion siRNA targeting MK gene can induce apoptosis via down-regulation of Bcl-2, mitochondrial depolarization and caspase-3 activation.
出处
《中国实验诊断学》
北大核心
2009年第4期427-429,共3页
Chinese Journal of Laboratory Diagnosis
基金
教育部重点项目(01058)
