Department of Fundamental Medicine, School of Biomedicine, Far Eastern Federal University, Vladivostok, Russia; Laboratory of Pharmacology, National Scientific Center of Marine Biology, Far East Branch of the Russian Academy of Sciences, Vladivostok, Russia; Medical Center, Far Eastern Federal University, Vladivostok, Russia.
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.
Int Rev Neurobiol. 2020;151:219-242. doi: 10.1016/bs.irn.2020.03.007. Epub 2020 May 13.
Glioblastoma multiforme is the most aggressive type of primary brain tumor in humans. Its invasive growth is associated with cluster of differentiation (CD)133 cancer stem cells (CSCs) and CD133 differentiated glioblastoma cells (DGCs) with aggressive phenotype, which are developed under the influence of transforming growth factor (TGF)-β. The present study aimed to compare the proteomes of CD133 CSCs and CD133 DGCs stimulated by TGF-β, as well as the expression levels of the main proteins responsible for activating the signaling pathway of receptor interactions with the extracellular matrix (ECM). The U87MG GBM cell line was used in this study. CSCs were extracted from gliomaspheres through magnetic-activated cell sorting based on the expression of CD133 (CD133); CD133 DCGs served as a control. CD133 DGCs of the U87-MG cell line were treated with 10ng/mL TGF-β1, and cell proliferation and migration were analyzed via real-time quantitative microscopy. High-performance liquid chromatography mass spectrometry was used for proteome analysis. The results revealed 589 proteins with significantly changes in expression among CD133 CSCs compared with those in CD133 DGCs (P<0.05). Bioinformatics analysis allowed to attribute 134 differentially expressed proteins to 15 signaling pathways; among these proteins, 14 were involved in signaling cascades associated with the interaction between CSCs and the ECM, and were upregulated >twofold, while four proteins activated this signaling cascade. TGF-β-stimulation increased the mobility, suppressed the proliferation and transformed the proteome profile of CD133 DGCs. Were identified 13 key proteins that activate the signaling pathway of receptor interaction with the ECM and three proteins activating this signaling pathway in CD133 DGCs which had the same values as those of CD133 CSCs. In conclusion, TGF-β increased the expression of proteins that activate the signaling pathway of receptor interaction with the ECM in CD133 DGCs to the level of those in CD133 CSCs.
多形性胶质母细胞瘤是人类最具侵袭性的原发性脑肿瘤。其浸润性生长与簇分化（CD）133 癌症干细胞（CSCs）和 CD133 分化的胶质母细胞瘤细胞（DGCs）有关，这些细胞具有侵袭表型，是在转化生长因子（TGF）-β的影响下发展而来的。本研究旨在比较 TGF-β刺激的 CD133 CSCs 和 CD133 DGCs 的蛋白质组，并比较激活细胞外基质（ECM）受体相互作用信号通路的主要蛋白的表达水平。本研究使用 U87MG GBM 细胞系。CSCs 是通过基于 CD133 表达的磁激活细胞分选从神经球中提取的（CD133）；CD133 DGC 作为对照。用 10ng/mL TGF-β1 处理 U87-MG 细胞系的 CD133 DGCs，通过实时定量显微镜分析细胞增殖和迁移。采用高效液相色谱-质谱联用技术进行蛋白质组分析。结果显示，与 CD133 DGCs 相比，CD133 CSCs 中有 589 种蛋白的表达发生了显著变化（P<0.05）。生物信息学分析将 134 种差异表达蛋白归因于 15 条信号通路；在这些蛋白中，有 14 种参与了与 CSCs 和 ECM 相互作用相关的信号级联反应，且上调了>2 倍，而有 4 种蛋白激活了这一信号级联反应。TGF-β刺激增加了 CD133 DGCs 的迁移能力，抑制了其增殖并改变了其蛋白质组谱。在 CD133 DGCs 中鉴定出 13 种激活细胞外基质受体相互作用信号通路的关键蛋白和 3 种激活该信号通路的蛋白，其值与 CD133 CSCs 相同。总之，TGF-β增加了 CD133 DGCs 中激活细胞外基质受体相互作用信号通路的蛋白表达水平，使其达到与 CD133 CSCs 相同的水平。
