摘要
目的 研究过表达巨噬细胞移动抑制因子（MIF）对宫颈癌SiHa细胞上皮间质转化（EMT）的影响作用.方法 通过构建重组真核表达载体构建pEGFP-N1-MIF,转染宫颈癌SiHa细胞（SiHa-pEGFP-N1-MIF组）,同时设空载体转染组（SiHa-pEGFP-N1组）和空白对照组（SiHa细胞组）,采用蛋白质印迹法（western blot W.B）检测SiHa细胞中MIF蛋白表达的变化,并检测转染前后SiHa细胞中EMT相关指标E-cadherin和Vimentin的表达变化.结果 pEGFP-N1-MIF转染SiHa细胞后MIF蛋白的表达量增加,高于对照组,差异有统计学意义（P＜0.05）;且过表达MIF基因后,与对照组相比,pEGFP-N1-MIF组SiHa细胞上皮标志物E-cadherin的蛋白表达水平下调（P＜0.05）,间叶标志物Vimentin的蛋白表达水平显著上调（P＜0.05）.结论 过表达MIF基因可促进宫颈癌SiHa细胞发生上皮-间质转化的能力.
Objective To investigate the effect of over-expressed macrophage migration inhibitory factor （MIF） on epithelial-mesenchymal transition （EMT） in human cervical carcinoma SiHa cells.Methods Recombinant eukaryotic expression plasmid liposome enhanced transfection of green fluorescent protein gene （pEGFP-N1）-MIF was constructed and then transfected into human cervical cancer SiHa cells.Experimental cells were classified into three groups （SiHa-pEGFP-N1-MIF,SiHa-pEGFP-N1,and SiHa）.Western blot was used to detect the expression of MIF protein,and the expressions of EMT-related markers such as E-cadherin and vimentin in SiHa cells were determined before and after transfection.Results The eukaryotic expression vector pEGFP-N1-MIF significantly increased the expression of MIF protein in SiHa cells （P 〈 0.05）,and after overexpression of MIF gene in SiHa cells,the expression of E-cadherin protein in SiHa-pEGFP-N1-MIF group was significantly lower than that in control groups （P 〈0.05）,while the expression of vimentin in SiHa-pEGFP-N1-MIF group was significantly higher than that in control groups （P 〈 0.05）.Conclusions Overexpression of MIF in cervical cancer SiHa cells can promote the EMT occurrence.
出处
《中国医师杂志》
CAS
2015年第4期484-486,490,共4页
Journal of Chinese Physician
基金
山西省科技攻关项目基金资助项目（20130313019-3）
