Effects of over-expression of macrophage migration inhibitory factor on the epithelial-mesenchymal transition in cervical cancer cells SiHa
Xiaofeng Shi, Wei Liu
Abstract
Xiaofeng Shi, Wei Liu
Abstract
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. Key words: Macrophage migration inhibitory factor; Epithelial-mesenchymal transition; Cervical cancer; E-cadherin; vimentin
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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. Key words: Macrophage migration inhibitory factor; Epithelial-mesenchymal transition; Cervical cancer; E-cadherin; vimentin
Key concepts: Vimentin, Epithelial–mesenchymal transition, Macrophage migration inhibitory factor, Immunocytochemistry, Transfection, Mesenchymal stem cell, Molecular biology, Chemistry