靶向MIF的siRNA对大肠癌细胞凋亡的影响  

The effect of the small interfering RNAs for macrophage migration inhibitory factor on apoptosis of colorectal cancer cell line

作　　者：吴礼浩[1] 徐慧鲜[1] 王丽京[2] 臧林泉[3] 何兴祥[1] 

机构地区：[1]广东药学院附属第一医院消化内科,广东广州510080 [2]广东药学院血管生物学研究所,广东广州510006 [3]广东药学院药科学院,广东广州510006

出　　处：《广东药学院学报》2011年第3期305-308,共4页Academic Journal of Guangdong College of Pharmacy

基　　金：广东省科学技术厅科技计划项目(2008A030201001);广东省自然科学基金(9251065005000001);广东省高等学校高层次人才项目(粤教师函[2010]79号)

摘　　要：目的研究化学合成的siRNA干扰巨噬细胞移动抑制因子(macrophage migration inhibitory factor,MIF)对大肠癌CT-26细胞凋亡的影响及其可能的机制。方法实验组用靶向MIF的siRNA处理CT-26细胞,对照组用非特异性的siRNA处理CT-26细胞,空白组不添加任何干扰剂。流式细胞仪检测细胞的凋亡情况,ELISA检测培养上清中MIF蛋白的含量,逆转录聚合酶链反应(RT-PCR)检测MIF、CD74、Caspases3、Caspases8 mRNA的表达,Western blot检测细胞内MIF、CD74、Caspases8蛋白的表达。结果化学合成的MIF siRNA处理大肠癌CT-26细胞24 h后,CT-26细胞凋亡率增加;实验组培养上清中MIF蛋白的含量与对照组和空白组比较显著下降;实验组MIF、CD74的mRNA表达量显著下降,Caspases3、Caspases8 mRNA的表达量明显升高;实验组细胞内MIF、CD74蛋白表达降低,而Caspases8蛋白表达升高。结论化学合成的MIF siRNA提高了CT-26细胞的凋亡率,其机制可能是靶向MIF的siRNA降低了MIF和CD74的表达,升高了Caspases3、Caspases8的表达。Objective To investigate the effective mechanisms of the small interfering RNAs（siRNA） for macrophage migration inhibitory factor（MIF） on the apoptosis of mouse colorectal cancer cell line,CT-26.Methods CT-26 cells were treated with specific MIF siRNA as the experimental group,treated with nonspecific siRNA as the control group,and treated without siRNA as the blank group.Cell apoptosis was determined by flow cytometry.The levels of MIF protein in culture supernatant were examined using ELISA.mRNA expression of MIF,CD74,Caspase3 and Caspase8 was detected by RT-PCR.The expression of MIF,CD74 and Caspase8 protein was determined using Western blot.Results MIF siRNA treatment remarkably promoted the apoptosis of CT-26 cells.Meanwhile,MIF siRNA inhibited mRNA and protein expression of MIF and CD74,but increased the expression of Caspase3 and Caspase8 mRNA and Caspase8 protein in CT-26 cells.Conclusion Treatment with MIF siRNA promotes the apoptosis of CT-26 cells,and the underlying mechanisms of MIF siRNA are related to suppression of MIF and CD74 gene expression and enhancement of Caspase3 and Caspase8 gene expression.

关 键 词：巨噬细胞移动抑制因子(MIF) SIRNA 大肠癌 细胞凋亡 

分 类 号：R735.34[医药卫生—肿瘤]