Department of Surgery (RMH), The University of Melbourne, Parkville VIC 3010, Australia.
Department of Surgery (RMH), The University of Melbourne, Parkville VIC 3010, Australia; Department of Neurosurgery, The Royal Melbourne Hospital, Parkville 3050, Australia.
Exp Cell Res. 2019 Jan 15;374(2):353-364. doi: 10.1016/j.yexcr.2018.12.010. Epub 2018 Dec 15.
Glioblastoma (GBM) tumor cells exhibit drug resistance and are highly infiltrative. GBM stem cells (GSCs), which have low proliferative capacity are thought to be one of the sources of resistant cells which result in relapse/recurrence. However, the molecular mechanisms regulating quiescent-specific tumor cell biology are not well understood. Using human GBM cell lines and patient-derived GBM cells, Oregon Green dye retention was used to identify and isolate the slow-cycling, quiescent-like cell subpopulation from the more proliferative cells in culture. Sensitivity of cell subpopulations to temozolomide and radiation, as well as the migration and invasive potential were measured. Differential expression analysis following RNAseq identified genes enriched in the quiescent cell subpopulation. Orthotopic transplantation of cells into mice was used to compare the in vivo malignancy and invasive capacity of the cells. Proliferative quiescence correlated with better TMZ resistance and enhanced cell invasion, in vitro and in vivo. RNAseq expression analysis identified genes involved in the regulation cell invasion/migration and a three-gene signature, TGFBI, IGFBP3, CHI3L1, overexpressed in quiescent cells which correlates with poor GBM patient survival.
胶质母细胞瘤（GBM）肿瘤细胞表现出耐药性并且具有高度浸润性。GBM 干细胞（GSCs）增殖能力低，被认为是耐药细胞的来源之一，导致复发/重现。然而，调节静止特异性肿瘤细胞生物学的分子机制尚不清楚。使用人 GBM 细胞系和患者来源的 GBM 细胞，使用 Oregon Green 染料保留来鉴定和分离培养中增殖能力较高的细胞中的慢循环、静止样细胞亚群。测量细胞亚群对替莫唑胺和辐射的敏感性以及迁移和侵袭潜能。RNAseq 后的差异表达分析确定了在静止细胞亚群中富集的基因。将细胞原位移植到小鼠中，用于比较细胞在体内的恶性程度和侵袭能力。体外和体内研究表明，增殖性静止与更好的 TMZ 耐药性和增强的细胞侵袭相关。RNAseq 表达分析确定了参与调节细胞侵袭/迁移的基因，以及在静止细胞中过表达的 TGFBI、IGFBP3、CHI3L1 三个基因标志物，与 GBM 患者的不良预后相关。
