靶向MIF的siRNA对大肠癌细胞增殖的影响  

The Effect of the Small Interfering Rnas for Macrophage Migration Inhibitory Factor on the Proliferation of Colorectal Cancer Cell Line

作　　者：徐慧鲜[1] 吴礼浩[1] 马伟钦[1] 杨荣娇[1] 王亚敏[1] 王丽京[2] 何兴祥[1] 

机构地区：[1]广东药学院附属第一医院消化内科,广州市510080 [2]广东药学院基础实验楼血管生物学研究所,广州市510006

出　　处：《中华全科医学》2013年第3期334-335,397,共3页Chinese Journal of General Practice

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项目(9251065005000001);广东省高等学校高层次人才项目(粤教师函[2010]79号);国家自然科学基金青年科学基金项目(61201437)

摘　　要：目的研究化学合成的靶向MIF的siRNA干扰MIF后,对大肠癌CT-26细胞增殖的影响并探讨其可能的机制。方法 MTT法检测细胞的增殖情况;ELISA检测培养上清液中MIF蛋白的含量;逆转录聚合酶链反应(RT-PCR)检测MIF、CD74 mRNA的表达;Western blot检测细胞内MIF和CD74蛋白的表达。结果实验组CT-26细胞的增殖与对照组和空白组相比受到明显抑制(P24 h;100 nmol=0.003),呈时间-计量依赖关系;实验组培养上清液中MIF蛋白的含量与对照组和空白组比较显著减少(P=0.02);实验组中MIF与CD74的mRNA表达量与对照组和空白组相比显著下降(PMIF=0.001;PCD74=0.001);实验组MIF、CD74蛋白表达与对照组和空白组相比降低(PMIF=0.006;PCD74=0.016)。结论化学合成的MIF siRNA抑制了CT-26细胞的增殖,其可能机制是MIF siRNA降低了CT-26细胞内MIF与CD74的表达。Objective To investigate the effects of the small interfering RNAs（siRNA） for macrophage migration inhibitory factor（MIF） on the proliferation of colorectal cancer cell lines and approach its probable mechanisms.Methods MTT assay was used to evaluate the inhibition ratio of the proliferation.MIF protein level on the cultivation supernatant were determined with ELISA.The mRNA expression of MIF and CD74 was detected by reverse transcription polymerase chain reaction（RT-PCR）.MIF and CD74 protein level were determined using Western blot.Results It was found that MIFsiRNA markedly inhibited the proliferation of CT-26 cell in a time-dose relationship（P24 h;100nmol=0.003）;MIFsiRNA also inhibited the expression of MIF protein level on the cultivation supernatant（P=0.02）,and the mRNA expression of MIF and CD74（PMIF=0.001;PCD74=0.001）.In addition,MIF and CD74 protein level reduced（PMIF=0.006;PCD74=0.016）.Conclusion MIFsiRNA inhibited the proliferation of CT-26 cell.The mechanism may be MIFsiRNA reduce the mRNA expression of MIF and CD74;and which reduced the protein expression of MIF and CD74.

关 键 词：巨噬细胞移动抑制因子 SIRNA 大肠癌 

分 类 号：R735.34[医药卫生—肿瘤] R730.21[医药卫生—临床医学]