PHAP1通过AKT信号通路调控脑胶质瘤细胞增殖作用的研究

Studies on the effect of PHAP1 on proliferation of glioma cells by regulating AKT signaling pathway

目的探讨人类白细胞抗原关联蛋白1(putative HLA-associated protein1,PHAP1)是否通过蛋白激酶B信号通路进而影响胶质瘤细胞的增殖。方法应用Western blotting检测PHAP1在胶质瘤组织中的表达水平。用CCK-8实验检测沉默或者过表达PHAP1对U251和U87细胞增殖能力的影响;用免疫印迹法检测沉默或者过表达PHAP1对AKT/p27/Stathmin信号分子的影响;裸鼠体内验证PHAP1对胶质瘤移植瘤生长的作用。结果PHAP1在胶质瘤组织呈高表达。CCK-8实验显示,沉默PHAP1后胶质瘤细胞U251和U87的增殖能力下降,而过表达PHAP1后U251和U87细胞的增殖能力增强。Western blotting检测证实沉默PHAP1后,AKT活化显著下降,从而导致下游p27表达水平增高和Stathmin蛋白表达水平降低,过表达PHAP1产生相反的效应。进一步裸鼠体内实验验证,沉默PHAP1抑制胶质瘤移植瘤的生长。结论PHAP1可以通过调控AKT/p27/Stathmin信号通路,从而促进胶质瘤的增殖。

Objective To study the mechanism of putative HLA-associated protein1(PHAP1)regulated proliferation of glioma cells through protein kinase B(AKT)signaling pathway.Methods Western blotting was used to detect the expression of PHAP1 in glioma tissues.The effects of down-regulation or over-expression of PHAP1 on the proliferation of U251 and U87 cells were detected by CCK-8 assay.The effects of down-regulation or over-expression of PHAP1 on AKT/p27/Stathmin signaling molecules were detected by Western blotting.The role of PHAP1 in transplantated glioma was verified by in vivo experiment.Results PHAP1 was highly expressed in glioma tissues.CCK-8 experiment showed that the proliferation of glioma cells U251 and U87 decreased after down-regulation of PHAP1,however,the proliferation of glioma cells U251 and U87 was enhanced after over-expression of PHAP1.Western blotting confirmed that silencing of PHAP1 significantly reduced phosphorylated AKT,thereby increasing the expression level of p27 and decreasing the expression level of Stathmin.However,overexpression of PHAP1 had opposite effect.Expreiments in nude mice demonstrated that silencing PHAP1 inhibited the growth of transplanted glioma.Conclusion PHAP1 can promote the proliferation of glioma by regulating AKT/p27/Stathmin signaling pathway.