【摘要】 目的研究血清应答因子(serum response factor,SRF)小干扰RNA(small interference RNA,siRNA)影响转化生长因子β1(transforming growth factor-β1,TGF-β1)介导的Eca-109食管癌细胞发生上皮间质转化(epithelial-mesenchymal transition,EMT)的作用机制。方法体外培养Eca-109食管癌细胞,实验分组为阴性对照siRNA组、TGF-β1+阴性对照siRNA组、TGF-β1+SRFsiRNA组。划痕实验检测细胞迁移能力;免疫细胞化学染色法检测E-钙黏蛋白(E-cadherin)的表达;蛋白质印迹法检测E-钙黏蛋白、SRF、N-钙黏蛋白(N-cadherin)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)蛋白的表达。结果与阴性对照siRNA组细胞迁移百分比(10.00±2.00)%相比较,TGF-β1+阴性对照siRNA组细胞迁移百分比为(50.67±4.73)%,迁移能力增强;与TGF-β1+阴性对照siRNA组相比较,TGF-β1+SRF-siRNA组细胞迁移百分比为(29.00±3.00)%,迁移能力下降,均差异有统计学意义(P<0.001)。与阴性对照siRNA组的E-钙黏蛋白(1.07±0.12)、N-钙黏蛋白(0.28±0.25)、SRF(0.25±0.06)、α-SMA(1.19±0.37)蛋白相比较,TGF-β1+阴性对照siRNA组E-钙黏蛋白(0.45±0.06)表达下调,而N-钙黏蛋白(3.27±0.67)、SRF(2.48±0.05)、α-SMA(4.23±0.53)蛋白表达上调(均P<0.001);与TGF-β1+阴性对照siRNA组相比较,TGF-β1+SRF-siRNA组E-钙黏蛋白(0.82±0.05)表达上调,N-钙黏蛋白(1.31±0.13)、SRF(1.46±0.16)、α-SMA(2.60±0.28)蛋白表达下调(均P<0.001)。结论基因沉默SRF能够抑制TGF-β1介导的食管癌细胞发生EMT。更多还原

【Abstract】 Objective To study the inhibitory effects of serum response factor(SRF)-small interference RNA(siRNA) on epithelial-mesenchymal transition(EMT) in Eca-109 cells induced by transforming growth factor-β1(TGF-β1).Methods Eca-109 cells were cultured and divided into 3 group,including negative control-siRNA(NC-siRNA),TGF-β1+NC-siRNA,and TGF-β1+SRF-siRNA.Cell migration was measured by cell scratch test.The expression of E-cadherin was observed by immunocytochemistry.The expression of E-cadherin,SRF,N-cadherin,and α-smooth muscle actin(α-SMA) were measured by western blotting.Results Compared with siRNA-NC group(10.00±2.00)%,cell migration-was increased in TGF-β1+siRNA-NC group(50.67±4.73)%,and the migration ability-was enhanced;compared with TGF-β1+siRNA-NC group,cell migration-was(29.00±3.00)% in TGF-β1+SRF-siRNA group,and the migration ability was decreased.Compared with the expressions of E-cadherin(1.07±0.12),N-cadherin(0.28±0.25),SRF(0.25±0.06),and α-SMA(1.19±0.37) in the negative control siRNA group,the expression of E-cadherin(0.45±0.06)in the TGF-β1+negative control siRNA group was down-regulated,while the expression of N-cadherin(3.27±0.67),SRF(2.48±0.05),and α-SMA(4.23±0.53) were up-regulated(P<0.001).Compared with TGF-β1+NC-siRNA group,the expression of E-cadherin(0.82±0.05) in the TGF-β1+SRF-siRNA group was up-regulated,and the expression of N-cadherin(1.31±0.13),SRF(1.46±0.16),and α-SMA(2.60±0.28) were down-regulated(P<0.001).Conclusion Knock-down of SRF can inhibit the EMT induced by TGF-β1 in Eca-109 cells.更多还原