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机构地区:[1]山西医科大学,太原030001 [2]山西医学科学院,山西大医院妇产科
出 处:《中国医师杂志》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)
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.
关 键 词:印迹法 蛋白质 巨噬细胞游走抑制因子/代谢/药理学 宫颈肿瘤/代谢/病理学/药物疗法 细胞转化 肿瘤/药物作用 上皮细胞/药物作用/病理学 间质细胞/药物作用/病理学
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