| 何喜,王志强,林韬,等.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[J].安徽医药,2020,24(9):1768-1771. |
| 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 |
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| DOI：10.3969/j.issn.1009-6469.2020.09.018 |
| 中文关键词: 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 |
| 英文关键词: Esophageal neoplasms/etiology Serum response factor Cadherins Actins Epithelial interstitial transforma- tion Small interference RNA |
| 基金项目:河北省科技计划项目( 17277749D) |
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| 摘要点击次数: 1524 |
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| 中文摘要: |
| 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+SRF- siRNA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 siR- NA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。 |
| 英文摘要: |
| 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，and α-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)，and α-SMA(1.19±0.37)in the negative control siRNA group，the expression of E-cadherin(0.45±0.06) GF-β1+ negative control siRNA group was down-regulated，while the expression of N-cadherin(3.27±0.67)，SRF(2.48±intheT，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-cad- herin.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±(0，0.16)，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. |
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