GSK-3β过表达促进人胶质瘤细胞增殖及侵袭

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

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pc DNA3.1(+)-GSK-3β(r GSK-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。

Objective To explore the expression level 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 astrocytes1800 were analyzed by RT-PCR and Western blotting. The constructed pc DNA3. 1（ ＋）-GSK-3β（ r GSK-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）. Conclusions GSK-3β,via PAX3,promotes cell proliferation and invasion of U87 glioma cells.