基础医学与临床 ›› 2015, Vol. 35 ›› Issue (3): 371-376.

• 研究论文 • 上一篇    下一篇

GSK-3β过表达促进人胶质瘤细胞增殖及侵袭

石歆   

  1. 河南省中医学院第一附属医院
  • 收稿日期:2014-05-12 修回日期:2014-10-25 出版日期:2015-03-05 发布日期:2015-03-03
  • 通讯作者: 石歆 E-mail:shixin_tcm@163.com

Overexpression of GSK-3β promotes the proliferation and invasion of human glioma cell

  • Received:2014-05-12 Revised:2014-10-25 Online:2015-03-05 Published:2015-03-03

摘要: 目的 分析糖原合酶激酶-3(Glycogen Synthase Kinase-3β,GSK-3β)在胶质瘤细胞中的表达水平,探讨GSK-3β对胶质瘤细胞增殖、侵袭的影响及其作用机制。方法 RT-PCR及Western blot分析GSK-3β在人正常星形胶质细胞1800及A172、T98G、U87和U251 4种胶质瘤细胞中的表达水平;将构建的pcDNA3.1(+)-GSK-3β(rGSK-3β)重组表达载体及PAX3 siRNA转染人胶质瘤细U87, MTT法检细胞活力,Transwell小室检测细胞侵袭能力。结果 与1800细胞相比,4种胶质瘤细胞中GSK-3β的表达水平明显增高(P<0.05)。GSK-3β过表达促进了U87细胞的增殖,同时加强了侵袭细胞的数目(P<0.05)。分析表明,GSK-3β重组体转染显著加强细胞中PAX3的水平(P<0.05);当用PAX3 siRNA转染沉默细胞中PAX3的水平后,GSK-3β过表达诱导的细胞的增殖及侵袭能力明显降低(P<0.05)。结论 GSK-3β在胶质瘤细胞中过表达,且有可能通过PAX3来调控胶质瘤细胞的增殖及侵袭。

关键词: GSK-3β, 胶质瘤, 细胞增殖, 细胞侵袭, PAX3

Abstract: Objective To explore the expression levels of Glycogen Synthase Kinase-3β (GSK-3β) in glioma cells and its function on cell proliferation and invasion. The underlying mechanism was also investigated. Methods The expression levels of GSK-3β in four glioma cell lines (A172, T98, U87, U251) and normal human astrocytes 1800 were analyzed by RT-PCR and Western blotting. The constructed pcDNA3.1(+)-GSK-3β(rGSK-3β)recombinant vector and PAX3 siRNA were transfected into U87 cells. Cell viability and invasion ability were detected by MTT and Transwell invasion assay, respectively. Results The GSK-3β expression levels were detectable in four glioma cell lines (P<0.05), but low in the control 1800 cells. Overexpression of GSK-3β in U87 cells increased rates of cell proliferation was assessed by MTT assay and Ki67 expression. The GSK-3β expression promoted cell invasion(P<0.05). Further analysis suggested that the enhanced PAX3 was responsible for GSK-3β-induced cell proliferation and invasion. These advantages were abolished by inactivation of PAX3 using specific PAX3 siRNA (P<0.05). Conclusion This research demonstrated that GSK-3β, via PAX3, promotes the rates of cell proliferation and invasion in U87 glioma cells.

Key words: GSK-3β, glioma, cell proliferation, cell invasion, PAX3