转化生长因子β对A431细胞上皮向间质转换相关分子表达的影响
Effects of transforming growth factor-β on the expressions of epithelial-to-mesenchymal transition-related molecules in A431 cells
摘要目的 研究转化生长因子β(TGF-β)对人表皮鳞状细胞癌A431细胞上皮向间质转换(EMT)相关分子表达的影响.方法 用含7.5 μg/L TGF-β处理A431细胞72 h后观察细胞形态,采用实时定量PCR和Western印迹分别在mRNA水平和蛋白水平检测EMT相关基因E钙黏着蛋白、N钙黏着蛋白、波形蛋白、β联蛋白表达水平的变化.未经TGF-β处理的A431细胞为对照组.结果 TGF-β处理后,A431细胞独立分散,呈梭形.实时定量PCR结果显示,E钙黏着蛋白表达量为未经TGF-β处理组的31.7%,N钙黏着蛋白、波形蛋白、β联蛋白表达量分别为未经处理组的2.475、11.340、2.615倍;Western印迹结果,N钙黏着蛋白、波形蛋白、β联蛋白表达量上升,E钙黏着蛋白表达量下降.结论 TGF-β可诱导A431细胞产生EMT现象,TGF-β表达量上升,可能参与表皮鳞状细胞癌发生侵袭、转移.
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abstractsObjective 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.
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