RNA干扰沉默MIF基因对宫颈癌SiHa细胞上皮细胞间质变的影响

[目的]探讨巨噬细胞移动抑制因子（MIF）对人宫颈癌SiHa细胞上皮间质转化（EMT）的影响。[方法]构建并鉴定重组真核表达质粒Genesil-MIF siRNA,转染人宫颈癌细胞SiHa细胞。RT-PCR检测各组细胞中MIF、E-cadherin、Vimentin mRNA表达水平。免疫细胞化学法组检测E-cadherin、Vimentin蛋白表达水平。[结果]重组质粒Genesil-MIF siRNA成功构建并转染至人宫颈癌SiHa细胞。RT-PCR法证实实验组细胞中E-cadherin mRNA相对高表达（P〈0.05）,而Vimentin、MIF mRNA相对低表达（P〈0.05）。免疫细胞化学法证实实验组细胞中E-cadherin蛋白表达水平升高,Vimentin蛋白表达水平明显降低。[结论]MIF沉默通过促进SiHa细胞中E-cadherin表达,抑制Vimentin表达从而抑制EMT发生。

Effects of Silencing Migration Inhibitory Factor Gene by RNA Interference on Expression of EMT-associated Protein in Human Cervical Carcinoma SiHa Cell

[Purpose] To investigate the effect of RNA interference（RNAi） targeting macrophage migration inhibitory factor（MIF） on epithelial-mesenchymal transition（EMT）-associated proteins in human cervical carcinoma SiHa cells.[Methods] Recombinant eukaryotic expression plasmid pGenesil-MIF siRNA was constructed and identified,then transfected into human cervical cancer cells SiHa using a liposome approach. Real-time PCR was used to detect the silencing efficacy of MIF mRNA and the expression of E-cadherin、Vimentin mRNA.The protein expression of E-cadherin and Vimentin was detected by immunocytochemistry. [Results] The recombinant plasmid pEGFP-N1-MIF was constructed successfully and transfected into SiHa cells. In experimental group,the mRNA expression of Vimentin decreased,whereas E-cadherin expression increased. Immunocytochemical staining showed that expression of E-cadherin increased,whereas Vimentin expression decreased. [Conclusion] RNA interference of MIF suppresses epithelialmesenchymal transition in SiHa cells by decreasing the expression of E-cadherin and increasing the expression of Vimentin.