目的：探讨靶向沉默Notch⁃1基因对滋养细胞JEG⁃3上皮-间质转化（EMT）过程和侵袭能力的影响及相关机制。方法设计合成Notch⁃1⁃siRNA干扰片段，稳定转染JEG⁃3细胞。采用Transwell小室侵袭实验检测滋养细胞侵袭能力的变化。采用Western blot检测转染后各组细胞Notch⁃1蛋白、E⁃cadherin、vimentin蛋白的表达和EMT转录调节子slug，snail，twist的表达水平变化。结果转染Notch⁃1⁃siRNA后，滋养细胞JEG⁃3中Notch⁃1蛋白的相对表达量明显下降；滋养细胞JEG⁃3的侵袭能力明显降低；JEG⁃3细胞中上皮细胞标志物E⁃cadherin表达显著升高，而间质细胞标志物vimentin表达显著下降；EMT转录调节因子slug、snail和twist的表达显著下降。结论靶向沉默Notch⁃1基因能够影响滋养细胞的浸润；Notch⁃1通过调节滋养细胞EMT过程降低其侵袭能力。%Objective To discuss the effect and related mechanism on epithelial⁃mesenchymal transition(EMT)process and invasive ability of the trophoblasts JEG⁃3 by targeted silencing of Notch⁃1 gene. Methods Notch⁃1⁃siRNA interference fragment was designed and synthesized to sta⁃bly transfect JEG⁃3 cells. Changes in the invasive ability of trophoblasts were detected by Transwell chamber invasion assay. The expressions of Notch⁃1 ,E⁃cadherin ,vimentin and EMT transcriptive regulators slug ,snail ,twist were analyzed in transfected trophoblasts by Western blot. Results The relative expression of Notch⁃1 protein in JEG⁃3 and the invasive ability of JEG⁃3 were significantly decreased after transfection with Notch⁃1⁃siRNA,the expression of the epithelial indictor E⁃cadherin was significantly increased while the mesenchymal indicator vimentin was de⁃creased with the reduction of EMT transcriptive regulators slug,snail and twist. Conclusion The invasion of trophoblasts was affected by targeted si⁃lencing of Notch⁃1 gene and Notch⁃1 can reduce the invasion of trophoblasts through influencing EMT process.
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