College of Biological Science and Engineering, Fuzhou University, Fuzhou, Fujian, China.
Department of Trauma Center & Emergency Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou, China.
Cell Signal. 2023 Jun;106:110656. doi: 10.1016/j.cellsig.2023.110656. Epub 2023 Mar 17.
Glioma stem cells (GSCs) are thought to be responsible for the initiation and progression of glioblastoma (GBM). GBM presents highly invasive growth with a very high recurrence rate, so it has become a clinical problem to be solved urgently. RNAseq demonstrates that thrombospondin 1 (THBS1) acts not only in the angiogenic core of glioma but also with a high degree of invasiveness and infiltration. Nevertheless, defects in the signaling pathway research lead to a poor prognosis in glioma patients. To investigate the relevant molecular mechanism and signal pathway of glioma stem cell behavior mediated by THBS1, U251 astroglioma cells and GSCs were taken as model cells for in vitro experiments. The biological effects of THBS1 on glioma proliferation, migration, and adhesion were evaluated using Cell Counting Kit-8(CCK8) assays, EdU incorporation assays, migration assays, Transwell assays, Western blotting, and RNAseq. We found that the knockout of the THBS1 gene by CRISPR/Cas9 promoted proliferation and migration in U251 cells and GSCs, as well as influencing cell cycle progression by regulating the TNF/MAPK/NF-κB and TGF-β/Smad signaling pathways. Moreover, U251 cells and GSCs showed different responses to THBS1 knockout, suggesting specific and potential targets for GSCs in signaling pathways mediated by THBS1.
神经胶质瘤干细胞（GSCs）被认为是导致胶质母细胞瘤（GBM）发生和发展的原因。GBM 呈现出高度侵袭性生长和极高的复发率，因此成为亟待解决的临床问题。RNAseq 表明，血小板反应蛋白 1（THBS1）不仅在神经胶质瘤的血管生成核心中起作用，而且具有高度的侵袭性和浸润性。然而，信号通路研究中的缺陷导致神经胶质瘤患者预后不良。为了研究 THBS1 介导的神经胶质瘤干细胞行为的相关分子机制和信号通路，我们以 U251 星形细胞瘤细胞和 GSCs 作为体外实验的模型细胞。通过 CCK8 检测、EdU 掺入检测、迁移检测、Transwell 检测、Western blot 和 RNAseq 评估 THBS1 对神经胶质瘤增殖、迁移和黏附的生物学效应。我们发现，CRISPR/Cas9 敲除 THBS1 基因促进了 U251 细胞和 GSCs 的增殖和迁移，并通过调节 TNF/MAPK/NF-κB 和 TGF-β/Smad 信号通路影响细胞周期进程。此外，U251 细胞和 GSCs 对 THBS1 敲除的反应不同，这表明在 THBS1 介导的信号通路中，GSCs 具有特定的潜在靶点。
