Li Rujun, Li Junyang, Sang Dongping, Lan Qing
Department of Neurosurgery, Second Affiliated Hospital of Soochow University, 1055 Sanxiang Road, Suzhou, 215004, Jiangsu, China.
J Neurooncol. 2015 Jan;121(1):83-9. doi: 10.1007/s11060-014-1610-3. Epub 2014 Sep 9.
The aim of this study is to determine whether phosphorylation of AKT could be effected by t-AUCB-induced p-Hsp27 and whether p-AKT inhibition sensitizes glioblastoma cells to t-AUCB, and to evaluate the effects of simultaneous inhibition of p-Hsp27 and p-AKT on t-AUCB treated glioblastoma cells. Cell growth was detected using CCK-8 assay; Caspase-3 activity assay kits and flow cytometry were used in apoptosis analysis; Western blot analysis was used to detect p-Hsp27 and p-AKT levels; RNA interference using the siRNA oligos of Hsp27 was performed to knockdown gene expression of Hsp27. All data were analyzed by the Student-Newman-Keul's test. We demonstrated that t-AUCB treatment induces AKT phosphorylation by activating Hsp27 in U251 and LN443 cell lines. Inhibition of AKT phosphorylation by AKT inhibitor IV sensitizes glioblastoma cells to t-AUCB, strengthens t-AUCB suppressing cell growth and inducing cell apoptosis. We also found inhibiting both p-Hsp27 and p-AKT synergistically strengthen t-AUCB suppressing cell growth. Thus, p-AKT induced by p-Hsp27 confers the apoptosis-resistance in t-AUCB-treated glioblastoma cells. Targeting p-Hsp27 and/or p-AKT may be a potential effective strategy for the treatment of glioblastoma.
本研究的目的是确定t-AUCB诱导的p-Hsp27是否会影响AKT的磷酸化，以及p-AKT抑制是否会使胶质母细胞瘤细胞对t-AUCB敏感，并评估同时抑制p-Hsp27和p-AKT对t-AUCB处理的胶质母细胞瘤细胞的影响。使用CCK-8法检测细胞生长；使用Caspase-3活性检测试剂盒和流式细胞术进行凋亡分析；使用蛋白质免疫印迹分析检测p-Hsp27和p-AKT水平；使用Hsp27的siRNA寡核苷酸进行RNA干扰以敲低Hsp27的基因表达。所有数据均采用Student-Newman-Keul检验进行分析。我们证明，在U251和LN443细胞系中，t-AUCB处理通过激活Hsp27诱导AKT磷酸化。AKT抑制剂IV抑制AKT磷酸化可使胶质母细胞瘤细胞对t-AUCB敏感，增强t-AUCB抑制细胞生长和诱导细胞凋亡的作用。我们还发现，同时抑制p-Hsp27和p-AKT可协同增强t-AUCB抑制细胞生长的作用。因此，p-Hsp27诱导的p-AKT赋予了t-AUCB处理的胶质母细胞瘤细胞抗凋亡能力。靶向p-Hsp27和/或p-AKT可能是治疗胶质母细胞瘤的一种潜在有效策略。
