抑制血清应答因子表达影响转化生长因子β1介导的食管癌上皮细胞-间质转化的作用研究  

The inhibitory effects of SRF⁃siRNA on EMT in Eca⁃109 cells

作　　者：何喜[1] 王志强[1] 林韬[1] 胡万宁[1] 张明明 HE Xi;WANG Zhiqiang;LIN Tao;HU Wanning;ZHANG Mingming(epartment of General Thoracic Surgery,Tangshan People’s Hospital,Tangshan,Hebei 063001,China)

机构地区：[1]唐山市人民医院胸外科,河北唐山063001

出　　处：《安徽医药》2020年第9期1768-1771,I0004,共5页Anhui Medical and Pharmaceutical Journal

基　　金：河北省科技计划项目(17277749D)。

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。Objective To study the inhibitory effects of serum response factor(SRF)⁃small interference RNA(siRNA)on epitheli⁃al⁃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⁃siR⁃NA.Cell migration was measured by cell scratch test.The expression of E⁃cadherin was observed by immunocytochemistry.The ex⁃pression of E⁃cadherin,SRF,N⁃cadherin,andα⁃smooth muscle actin(α⁃SMA)were measured by western blotting.Results Com⁃pared 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⁃siR⁃NA 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),andα⁃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),andα⁃SMA(4.23±0.53)were up⁃regulated(P<0.001).Compared with TGF⁃β1+NC⁃siRNA group,the expression of E⁃cad⁃herin(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),andα⁃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.

关 键 词：食管肿瘤/病因学 血清反应因子 钙黏着糖蛋白类 肌动蛋白类 上皮-间质转化 小干扰RNA 

分 类 号：R735.1[医药卫生—肿瘤]