Cell Polarity, Migration and Cancer Unit, Université Paris-Cité, Institut Pasteur, Équipe Labellisée Ligue Contre le Cancer.
Cell Polarity, Migration and Cancer Unit, Université Paris-Cité, Institut Pasteur, Équipe Labellisée Ligue Contre le Cancer;
J Vis Exp. 2022 Jul 29(185). doi: 10.3791/64093.
With a dismal median survival time in real populations-between 6 to 15 months-glioblastoma (GBM) is the most devastating malignant brain tumor. Treatment failure is mainly due to the invasiveness of GBM cells, which speaks for the need for a better understanding of GBM motile properties. To investigate the molecular mechanism supporting GBM invasion, new physiological models enabling in-depth characterization of protein dynamics during invasion are required. These observations would pave the way to the discovery of novel targets to block tumor infiltration and improve patient outcomes. This paper reports how an orthotopic xenograft of GBM cells in the zebrafish brain permits subcellular intravital live imaging. Focusing on microtubules (MTs), we describe a procedure for MT labeling in GBM cells, microinjecting GBM cells in the transparent brain of 3 days post fertilization (dpf) zebrafish larvae, intravital imaging of MTs in the disseminating xenografts, altering MT dynamics to assess their role during GBM invasion, and analyzing the acquired data.
在真实人群中，胶质母细胞瘤（GBM）的中位生存时间惨淡，只有 6 到 15 个月，是最具破坏性的恶性脑肿瘤。治疗失败主要是由于 GBM 细胞的侵袭性，这说明了需要更好地了解 GBM 的运动特性。为了研究支持 GBM 侵袭的分子机制，需要新的生理模型来深入描述侵袭过程中蛋白质动力学。这些观察结果将为发现阻止肿瘤浸润和改善患者预后的新靶点铺平道路。本文报告了在斑马鱼大脑中进行 GBM 细胞原位异种移植如何实现细胞内亚活体成像。本文聚焦于微管（MTs），描述了一种在 GBM 细胞中标记 MT 的方法，即将 GBM 细胞微注射到 3 天受精后（dpf）斑马鱼幼虫的透明脑中，对传播性异种移植中的 MT 进行活体成像，改变 MT 动力学以评估其在 GBM 侵袭过程中的作用，并分析所获得的数据。
