CAJ | 학술논문

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.
목적 탐토과표체거서세포이동억제인자(MIF)대자궁경암SiHa세포발생상피간질전화 (EMT)적영향.방법 응용기인전염방법장중조질립pEGFP-N1-MIF전입SiHa세포,구건과표체MIF적SiHa세포;용실시형광취합매련반응(RT-PCR)급면역세포화학방법분별검측각조세포중E-cadherin、vimentin mRNA급단백적표체수평.결과 RT-PCR검측결과현시,전염pEGFP-N1-MIF적세포중MIF mRNA상대표체량승고(F=2950.278,P＜ 0.01);전염pEGFP-NI-MIF적세포중vimentin적mRNA고우각대조조,이E-cadherin적mRNA저우각대조조(F치분별위2 135.048,1 893.563,균P＜0.01);전염pEGFP-N 1-MIF적세포중vimentin적단백표체수평고우각대조조,이E-cadherin적단백표체수평저우각대조조(F치분별위2 348.021,1 789.421,균P＜ 0.01).결론 과표체MIF촉진SiHa세포중vimentin표체,억제E-cadherin표체,설명과표체MIF촉진자궁경암SiHa세포발생상피간질전화.
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.