【摘 要】
：
本文旨在探讨敲低鞘氨醇激酶-1(sphingosine kinase-1,SPHK1)对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞增殖、周期和凋亡的影响,并探讨其可能机制.用定量逆转录聚合酶链反应(qRT-PCR)检测人胚肺成纤维细胞系(MRC-5)和4种NSCLC细胞中SPHK1 mRNA表达水平.利用SPHK1-shRNA和相应的阴性对照转染A549和H1299细胞.用CCK8法测定细胞增殖,用AnnexinV-FITC/PI双重染色试剂盒评价细胞凋亡,用细胞
【机 构】
：
广西壮族自治区人民医院临床肿瘤中心,南宁530021;广西壮族自治区人民医院病理科,南宁530021;广西壮族自治区人民医院科研部,南宁530021
论文部分内容阅读
本文旨在探讨敲低鞘氨醇激酶-1(sphingosine kinase-1,SPHK1)对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞增殖、周期和凋亡的影响,并探讨其可能机制.用定量逆转录聚合酶链反应(qRT-PCR)检测人胚肺成纤维细胞系(MRC-5)和4种NSCLC细胞中SPHK1 mRNA表达水平.利用SPHK1-shRNA和相应的阴性对照转染A549和H1299细胞.用CCK8法测定细胞增殖,用AnnexinV-FITC/PI双重染色试剂盒评价细胞凋亡,用细胞周期检测试剂盒测定细胞周期分布,用JC-1线粒体膜电位测定试剂盒检测线粒体膜电位,用蛋白质印迹法检测细胞周期和线粒体凋亡途径相关蛋白、MEK/ERK信号通路的蛋白表达水平.结果显示,NSCLC细胞SPHK1 mRNA表达水平高于MRC-5细胞.SPHK1-shRNA显著抑制A549和H1299细胞的增殖,并促进线粒体途径的细胞凋亡,将细胞周期阻滞在G0/G1期;与对照组相比,SPHK1-shRNA组MEK和ERK蛋白磷酸化水平显著下调.MEK/ERK抑制剂可抑制A549和H1299细胞增殖,并促进细胞凋亡.上述结果提示,SPHK1敲低可能通过抑制MEK/ERK信号通路来抑制NSCLC的增殖和促进线粒体途径的细胞凋亡.
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