过表达巨噬细胞移动抑制因子对子宫颈癌SiHa细胞上皮间质转化的影响

目的 探讨过表达巨噬细胞移动抑制因子（MIF）对子宫颈癌SiHa细胞发生上皮间质转化 （EMT）的影响.方法 应用基因转染方法将重组质粒pEGFP-N1-MIF转入SiHa细胞,构建过表达MIF的SiHa细胞;用实时荧光聚合酶链反应（RT-PCR）及免疫细胞化学方法分别检测各组细胞中E-cadherin、vimentin mRNA及蛋白的表达水平.结果 RT-PCR检测结果显示,转染pEGFP-N1-MIF的细胞中MIF mRNA相对表达量升高（F=2950.278,P＜ 0.01）;转染pEGFP-NI-MIF的细胞中vimentin的mRNA高于各对照组,而E-cadherin的mRNA低于各对照组（F值分别为2 135.048,1 893.563,均P＜0.01）;转染pEGFP-N 1-MIF的细胞中vimentin的蛋白表达水平高于各对照组,而E-cadherin的蛋白表达水平低于各对照组（F值分别为2 348.021,1 789.421,均P＜ 0.01）.结论 过表达MIF促进SiHa细胞中vimentin表达,抑制E-cadherin表达,说明过表达MIF促进子宫颈癌SiHa细胞发生上皮间质转化.

Effects of over-expression of macrophage migration inhibitory factor on the epithelial-mesenchymal transition in cervical cancer cells SiHa

Objective To investigate the impact of migration inhibitory factor （MIF） over-expression on the epithelial to mesenchymal transition in human cervical cancer SiHa cells.Methods Recombinant plasmid pEGFP-N1-MIF was transfected into SiHa cells,and then of MIF mRNA relative quantitative expression was tested by RT-PCR.The mRNA and protein expression level of E-cadherin and vimentin were detected by RT-PCR and immunocytochemistry,respectively.Results RT-PCR results showed that MIF mRNA expression quantity in experimental group was higher than that in control group cells （F =2 950.278,P 〈 0.01）.In MIF-overexpressing SiHa cells,vimentin mRNA was increased and E-cadherin mRNA was decreased determined by RT-PCR （Fvalues were 2 135.048,1 893.563,P〈 0.01）.Immunocytochemistry results showed that vimentin expression quantity in experimental group cells were higher than that in control group cells,however,E-cadherin expression quantity was lower than that in control group cells （F values were 2 348.021,1 789.421,P 〈 0.01）.Conclusions The over-expression of MIF gene can significantly up-regulate the expression of vimentin,and down-regulate the expression of E-cadherin.Consequently,MIF over-expression induces epithelial to mesenchymal transition in human cervical cancer SiHa cells.