Effects of transforming growth factor-β on the expressions of epithelial-to-mesenchymal transition-related molecules in A431 cells
Yuanyuan Guo, Bin Peng, Guiqiong Xiang, Songmei Geng
Abstract
Yuanyuan Guo, Bin Peng, Guiqiong Xiang, Songmei Geng
Abstract
Objective To evaluate the effect of transforming growth factor-β (TGF-β) on the expressions of epithelial-to-mesenchymal transition (EMT)-related molecules in human A431 epidermal squamous cell carcinoma cells.Methods Cultured A431 cells were classified into two groups:TGF-β group treated with TGF-β of 7.5 μg/L for 72 hours,and control group remaining untreated.Subsequently,cell morphology was observed under an inverted microscope,and real time-PCR and Western blot were performed to quantify the mRNA and protein expressions of EMT-related molecules including E-cadherin,N-cadherin,vimentin and β-catenin respectively.Results After TGF-β treatment,the cells became dispersed and spindle in shape.The mRNA expression levels of E-cadherin,Ncadherin,vimentin,and β-catenin in TGF-β group were 0.317,2.475,11.340 and 2.615 folds those in the control group respectively.Western blot showed a marked increase in the expression of N-cadherin,vimentin,and β-catenin proteins but a notable decrease in that of E-cadherin in the TGF-β group compared with the control group.Conclusions TGF-β can induce EMT in A431 cells,and increased expression of TGF-β may contribute to the invasion and metastasis of SCC. Key words: Cell line, tumor; Transforming growth factor beta; Epithelial-to-mesenchymal transition
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Objective To evaluate the effect of transforming growth factor-β (TGF-β) on the expressions of epithelial-to-mesenchymal transition (EMT)-related molecules in human A431 epidermal squamous cell carcinoma cells.Methods Cultured A431 cells were classified into two groups:TGF-β group treated with TGF-β of 7.5 μg/L for 72 hours,and control group remaining untreated.Subsequently,cell morphology was observed under an inverted microscope,and real time-PCR and Western blot were performed to quantify the mRNA and protein expressions of EMT-related molecules including E-cadherin,N-cadherin,vimentin and β-catenin respectively.Results After TGF-β treatment,the cells became dispersed and spindle in shape.The mRNA expression levels of E-cadherin,Ncadherin,vimentin,and β-catenin in TGF-β group were 0.317,2.475,11.340 and 2.615 folds those in the control group respectively.Western blot showed a marked increase in the expression of N-cadherin,vimentin,and β-catenin proteins but a notable decrease in that of E-cadherin in the TGF-β group compared with the control group.Conclusions TGF-β can induce EMT in A431 cells,and increased expression of TGF-β may contribute to the invasion and metastasis of SCC. Key words: Cell line, tumor; Transforming growth factor beta; Epithelial-to-mesenchymal transition
Key concepts: Vimentin, Epithelial–mesenchymal transition, Transforming growth factor, A431 cells, Cadherin, Cell culture, Epidermal growth factor, Western blot