摘要
目的探讨磷酸甘油酸激酶1(PGK1)对β-catenin的调控作用及对胶质细胞瘤侵袭性的影响。方法以siRNA及过表达载体干扰胶质瘤U87细胞中PGK1表达水平,并经Western Blot及qRT-PCR方法验证;通过荧光双染检测PGK1与β-catenin的共定位表达情况,观察PGK1对β-catenin表达的调控作用,并检测不同PGK1与β-catenin表达情况下,U87细胞的生长增殖、侵袭能力及迁移能力的变化。在不同PGK1表达水平的U87细胞中,经Western Blot检测不同磷酸化位点的β-catenin表达水平。结果PGK1 siRNA过表达载体转染后,U87细胞生长活性、侵袭和迁移细胞数随之出现相应下降或升高。PGK1可影响β-catenin的表达,荧光双染结果提示二者共定位区域有同向变化。PGK1可影响β-catenin磷酸化活性形式的表达。使用β-catenin抑制剂(XAV 939)处理胶质瘤细胞后,对上游PGK1表达无影响,但细胞生长活性降低,侵袭和迁移细胞数减少。结论PGK1可影响胶质瘤细胞中β-catenin的表达,二者可促进胶质瘤细胞增殖、迁移及侵袭生长,其机制或许与PGK1所致β-catenin磷酸化有关。
Objective To investigate the regulation of phosphoglycerate kinase 1(PGK1)onβ-catenin and its effect on the invasiveness of glioma.Methods SiRNA and overexpression vector were used to interfere with the expression level of PGK1 in glioma U87 cells,which was verified by Western blot and qRT-PCR.The co-localization expression of PGK1 andβ-catenin was detected by fluorescence double staining.The regulation of PGK1 on the expression ofβ-catenin was observed,and the changes of growth,proliferation,invasion and migration of U87 cells under different expression of PGK1 andβ-catenin were detected.In U87 cells with different PGK1 expression levels,the expression levels ofβ-catenin at different phosphorylation sites were detected by Western blot.Results After transfection with PGK1 siRNA overexpression vector,the growth activity,invasion and migration of U87 cells decreased or increased accordingly.PGK1 could affect the expression ofβ-catenin.The results of fluorescence double staining suggested that the co localization region of PGK1 andβ-catenin had the same direction change.PGK1 could affect the expression of phosphorylated active form ofβ-catenin.The treatment of glioma cells withβ-catenin inhibitor(XAV939)had no effect on the expression of upstream PGK1,but the cell growth activity decreased and the number of invasive and migratory cells decreased.Conclusions PGK1 can affect the expression ofβ-catenin in glioma cells.They can promote the proliferation,migration,invasion and growth of glioma cells.The mechanism may be related to the phosphorylation ofβ-catenin induced by PGK1.
作者
阎华
刘宇阳
陈永严
肖红
刘宏毅
YAN Hua;LIU Yu-yang;CHEN Yong-yan(Department of Neurosurgery,The Affiliated Brain Hospital of Nanjing Medical University,Nanjing 210029,China)
出处
《临床神经外科杂志》
2022年第2期135-141,共7页
Journal of Clinical Neurosurgery
基金
国家自然科学基金项目(81972350)
南京市卫健委基金资助项目(YKK15105)。