Department of Neurosurgery, Cangzhou Central Hospital, Xinhua West Road, Cangzhou, 061000, Hebei, China.
Department of Emergency, Cangzhou Central Hospital, Xinhua West Road, Cangzhou, 061000, Hebei, China.
J Mol Neurosci. 2019 Nov;69(3):391-398. doi: 10.1007/s12031-019-01368-y. Epub 2019 Jul 2.
Glioblastoma (GBM) is the most aggressive astrocytoma. Despite maximum treatment, the GBM usually recurs and the patient survival is poor. Thus, understanding the molecular mechanism of GBM progression will be meaningful to ameliorate this situation. In this study, collapsin response mediator protein 2 (CRMP2) and Ubc9 protein levels were evaluated in three GBM cell lines. Sumoylated CRMP2 were enriched and immunoprecipitated using SUMO1 and IgG antibodies. CRMP2-K374A mutant was generated by site-direct mutagenesis. All indicated constructs were transfected into GL15 cells, and the corresponding proliferation-promoting effect was assessed through cell proliferation ratio. The t-CSM peptide was used to disturb Ubc9-CRMP2 interaction. CRMP2 is expressed in all tested GBM cell lines. The Ubc9 protein levels are positively correlated with CRMP2 level, and both can promote GBM cell proliferation. Blocking CRMP2 SUMOylation through SUMOylation-incompetent mutant or small peptide suppresses CRMP2-induced GBM cell proliferation. This study demonstrates that the CRMP2 SUMOylation exists widely in GBM cells and drives glioblastoma proliferation. CRMP2 SUMOylation inhibition can significantly suppress GBM proliferation in vitro.
胶质母细胞瘤（GBM）是最具侵袭性的星形细胞瘤。尽管采用了最大程度的治疗，GBM 通常还是会复发，患者的生存状况较差。因此，了解 GBM 进展的分子机制对于改善这种情况将具有重要意义。在这项研究中，评估了三种 GBM 细胞系中 collapsin 反应介质蛋白 2（CRMP2）和 Ubc9 蛋白的水平。使用 SUMO1 和 IgG 抗体富集和免疫沉淀 SUMO 化的 CRMP2。通过定点突变生成 CRMP2-K374A 突变体。将所有构建体转染到 GL15 细胞中，并通过细胞增殖比评估相应的促进增殖作用。使用 t-CSM 肽干扰 Ubc9-CRMP2 相互作用。CRMP2 在所有测试的 GBM 细胞系中表达。Ubc9 蛋白水平与 CRMP2 水平呈正相关，两者均可促进 GBM 细胞增殖。通过 SUMOylation 失活突变体或小肽阻断 CRMP2 SUMOylation 可抑制 CRMP2 诱导的 GBM 细胞增殖。本研究表明，CRMP2 SUMOylation 广泛存在于 GBM 细胞中，并驱动胶质母细胞瘤增殖。CRMP2 SUMOylation 抑制可显著抑制体外 GBM 增殖。
