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PC4与丝氨酸/精氨酸富集剪接因子1相互作用蛋白1的RNA干扰抑制U87胶质瘤细胞的侵袭和迁移

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

作者信息

Xiang Wei, Qi Song-Tao, Liu Ya-Wei, Li He-Zhen, Zhou Qiang, Yi Guo-Zhong, Chen Zi-Yang, Yan Le

机构信息

Department of Neurosurgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. E-mail:

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2016 Jun;36(6):802-6.

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.

CONCLUSION

s 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.

摘要

目的

研究小干扰RNA(siRNA)介导的PC4和丝氨酸/精氨酸丰富剪接因子1相互作用蛋白1(PSIP1)沉默对人胶质瘤U87细胞侵袭和迁移能力的影响。

方法

通过脂质体将化学合成的靶向PSIP1基因的siRNA转染至U87细胞,采用实时定量聚合酶链反应(PCR)检测基因沉默效果。分别采用Transwell小室实验和划痕实验评估转染细胞侵袭和迁移能力的变化。采用蛋白质免疫印迹法分析N-钙黏蛋白、β-连环蛋白和转录因子Slug的表达情况。

结果

转染PSIP1 siRNA后,U87细胞中PSIP1的mRNA和蛋白水平显著降低(P<0.0001)。U87细胞中PSIP1表达下调导致细胞侵袭和迁移能力显著受到抑制(P<0.01),同时N-钙黏蛋白、β-连环蛋白和Slug的表达也降低。

结论

PSIP1沉默可损害胶质瘤细胞的侵袭和迁移能力,并降低N-钙黏蛋白、β-连环蛋白和Slug的表达,提示PSIP1可能通过经典的Wnt/β-连环蛋白信号通路调节Slug,从而调控上皮-间质转化,促进胶质瘤细胞的侵袭和迁移。

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