Department of Fundamental Medicine, School of Biomedicine, Far Eastern Federal University, Vladivostok, Russia; Laboratory of Oncoproteomics, Institute of Carcinogenesis, N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia, Moscow, Russia.
Laboratory of Oncoproteomics, Institute of Carcinogenesis, N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia, Moscow, Russia.
Int Rev Neurobiol. 2020;151:185-200. doi: 10.1016/bs.irn.2020.03.006. Epub 2020 May 13.
Glioblastoma multiforme (GBM) is the most aggressive primary glial brain tumor. The prognosis for GBM patients is not favorable, with the median survival time being 15 months. Its treatment resistance is associated with GBM cell population having cancer stem cells (CSCs). Wnt/β-catenin signaling pathway is a strategically important molecular mechanism, providing proliferation of stem cells of all types. This study compares the expression levels of signaling pathway proteins in CD133(+) CSCs and CD133(-) differentiated glioblastoma cells (DGCs).
the present study used U-87MG cells of human glioblastoma, the material was tested for mycoplasma contamination. High-performance liquid chromatography (HPLC) mass spectrometry was used for proteome analysis. Biological and molecular functions, signaling pathways and protein-protein interactions were analyzed using free-access databases: PubMed, PANTHER, Gene Ontology, Swiss-Prot and KEGG. Protein-protein interactions (PPIs) were analyzed using the STRING database (version 10).
There were identified 589 proteins with significantly changed expression in CD133+ CSCs, as compared with CD133-DGCs (P<0.05). Bioinformatics analysis allowed to attribute 134 differentially expressed proteins to 16 signaling pathways. A significant increase in expression of eight Wnt signaling pathway proteins (APC, CSNK1E, CSNK1A, CSNK2A2, CSNK2B, CTNNB1, DVL1, RUVBL) was detected, as well as four proteins of the non-canonical Wnt pathway-RHOA, ROCK2, RAC2, DAAM1. Special attention should be paid to β-catenin (CTNNB1) with more than 13.98-fold increase of expression in CSCs and Disheveled-associated activator of morphogenesis 1 (DAAM1) with 6.15-fold higher upregulation level.
proteins of Wnt/β-catenin signaling cascade are a prospective target for regulating CSCs activity.
多形性胶质母细胞瘤（GBM）是最具侵袭性的原发性神经胶质瘤。GBM 患者的预后不佳，中位生存时间为 15 个月。其治疗耐药性与 GBM 细胞群体中的癌症干细胞（CSC）有关。Wnt/β-catenin 信号通路是一种具有战略重要性的分子机制，为各种类型的干细胞增殖提供支持。本研究比较了 CD133(+) CSC 和 CD133(-)分化神经胶质瘤细胞（DGC）中信号通路蛋白的表达水平。
本研究使用人胶质母细胞瘤 U-87MG 细胞，对其进行支原体污染检测。采用高效液相色谱-质谱联用（HPLC-MS）进行蛋白质组分析。利用免费数据库：PubMed、PANTHER、基因本体论（GO）、Swiss-Prot 和 KEGG 分析生物和分子功能、信号通路和蛋白质-蛋白质相互作用。使用 STRING 数据库（版本 10）分析蛋白质-蛋白质相互作用（PPIs）。
在 CD133+CSC 与 CD133-DGC 相比，发现有 589 种蛋白的表达有显著差异（P<0.05）。生物信息学分析将 134 种差异表达蛋白归因于 16 种信号通路。检测到 8 种 Wnt 信号通路蛋白（APC、CSNK1E、CSNK1A、CSNK2A2、CSNK2B、CTNNB1、DVL1、RUVBL）和 4 种非经典 Wnt 通路蛋白（RHOA、ROCK2、RAC2、DAAM1）的表达显著增加。应特别注意 CSC 中β-连环蛋白（CTNNB1）表达增加超过 13.98 倍和形态发生激活蛋白 1（DAAM1）表达增加 6.15 倍。
Wnt/β-catenin 信号级联中的蛋白是调节 CSC 活性的有前途的靶点。
