南方医科大学学报 ›› 2016, Vol. 36 ›› Issue (06): 802-.

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siRNA干扰相互作用蛋白1表达抑制胶质瘤细胞的侵袭和迁移

向伟,漆松涛,刘亚伟,李和珍,周强,易国仲,陈子阳,严乐   

  • 出版日期:2016-06-20 发布日期:2016-06-20

RNA interference of PC4 and SFRS1 interacting protein 1 inhibits invasion and migration of U87 glioma cells

  • Online:2016-06-20 Published:2016-06-20

摘要: 目的探讨小干扰RNA(siRNA)干扰降低PC4和SFRS1相互作用蛋白1(PSIP1)的表达对人胶质瘤U87细胞侵袭和迁移 的影响并初步探讨其机制。方法采用化学合成靶向PSIP1的siRNA,脂质体法转染U87细胞,用Real-time PCR检测RNA干 扰效率;Transwell侵袭实验测细胞侵袭能力;划痕实验检测其迁移能力;干扰PSIP1表达后,Western blot 检测间质细胞来源的 N-cadherin、β-catenin蛋白和转录因子Slug的表达。结果PSIP1-siRNA 转染胶质瘤U87细胞后,PSIP1的mRNA水平及蛋白 水平明显下降(P<0.001),U87 细胞侵袭和迁移能力下降,与阴性转染组及未转染组相比差异有统计学意义(P<0.001,P< 0.001),Western blot 显示,间质细胞来源的N-cadherin、β-catenin 蛋白表达量降低,同时与上皮间质转化(EMT)相关的转录因 子Slug 的表达降低。结论在胶质瘤中,抑制PSIP1 的表达可抑制肿瘤细胞的侵袭和迁移,N-cadherin、β-catenin 蛋白和转录 因子Slug的表达量降低,提示PSIP1 可通过调控经典Wnt/β-catenin 信号通路来调节Slug 的表达,进而调控EMT,从而参与胶 质瘤U87 细胞的侵袭及迁移。

Abstract: Objective To investigate the effect of small interfering RNA (siRNA)-mediated silencing of PC4 and SFRS1 interacting protein 1 (PSIP1) on invasion and migration of human glioma U87 cells. Methods Chemically synthesized siRNA targeting PSIP1 gene was transfected into U87 cells via lipofectamine, and the gene silencing effect was determined using real-time PCR. The changes in the invasion and migration abilities of the transfected cells were assessed with Transwell assay and wound healing assay, respectively. Western blotting was used to analyze the expression of N-cadherin, β-catenin and the transcription factor Slug. Results The mRNA and protein level of PSIP1 was significantly reduced in U87 cells after transfection with PSIP1 siRNA (P<0.0001). PSIP1 knockdown in U87 cells resulted in significant suppression of cell invasion and migration abilities (P<0.01) and also reduced N-cadherin, β-catenin and Slug expressions. Conclusions Silencing of PSIP1 impairs the invasion and migration abilities of glioma cells and lowers the expressions of N-cadherin, β-catenin and Slug, suggesting that PSIP1 may regulate Slug by classical Wnt/β-catenin signaling pathway to modulate epithelial-mesenchymal transition and promote the invasion and migration of glioma cells.