Abstract
Glioblastoma multiforme (GBM, WHO grade IV astrocytoma) is the most common and lethal primary brain tumor in adults, with an average survival of slightly more than 1 year after initial diagnosis. GBMs display significant heterogeneity within the tumor mass, among which a subpopulation of cells called glioma-initiating cells (GICs) is responsible for tumorigenesis and resistance to conventional therapies. Therefore, understanding the mechanism underlying the biological properties of GICs would help develop better therapies to target this population for GBM treatment. This protocol provides detailed procedures to isolate GICs and non-GICs from patient’s specimen and glioma xenografts, which serves as the first step for the biological studies of GICs. Upon separation of GICs and non-GICs, a series of studies, such as expression profiling and functional screen, etc., can be performed to identify signal pathways responsible for the malignant nature of GICs. Besides, translational studies can also be conducted to examine drug responses of GICs. In sum, isolation of GICs with reliable methods will provide the basis for the further biological studies.
The authors have declared that no conflict of interest exists.
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References
Ostrom QT, et al. CBTRUS statistical report: primary brain and central nervous system tumors diagnosed in the United States in 2007–2011. Neuro Oncol. 2014;16 Suppl 4:p. iv1–63.
Singh SK et al. Identification of human brain tumour initiating cells. Nature. 2004;432(7015):396–401.
Bao S et al. Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature. 2006;444(7120):756–60.
Bao SD et al. Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature. 2006;444(7120):756–60.
Eramo A et al. Chemotherapy resistance of glioblastoma stem cells. Cell Death Differ. 2006;13(7):1238–41.
Chen J et al. A restricted cell population propagates glioblastoma growth after chemotherapy. Nature. 2012;488(7412):522–6.
Wang H et al. miR-33a promotes glioma-initiating cell self-renewal via PKA and NOTCH pathways. J Clin Investig. 2014;124(10):4489–502.
Hu J et al. MiR-215 is induced post-transcriptionally via HIF-Drosha complex and mediates glioma-initiating cell adaptation to hypoxia by targeting KDM1B. Cancer Cell. 2016;29(1):49–60.
Ernst A et al. Genomic and expression profiling of glioblastoma stem cell-like spheroid cultures identifies novel tumor-relevant genes associated with survival. Clin Cancer Res. 2009;15(21):6541–50.
Schraivogel D et al. CAMTA1 is a novel tumour suppressor regulated by miR-9/9*in glioblastoma stem cells. EMBO J. 2011;30(20):4309–22.
Yin AH et al. AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood. 1997;90(12):5002–12.
Peichev M et al. Expression of VEGFR-2 and AC133 by circulating human CD34(+) cells identifies a population of functional endothelial precursors. Blood. 2000;95(3):952–8.
Uchida N et al. Direct isolation of human central nervous system stem cells. Proc Natl Acad Sci U S A. 2000;97(26):14720–5.
Medema JP. Cancer stem cells: the challenges ahead. Nat Cell Biol. 2013;15(4):338–44.
Beier D et al. CD133(+) and CD133(-) glioblastoma-derived cancer stem cells show differential growth characteristics and molecular profiles. Cancer Res. 2007;67(9):4010–5.
Chen R et al. A hierarchy of self-renewing tumor-initiating cell types in glioblastoma. Cancer Cell. 2010;17(4):362–75.
Son MJ et al. SSEA-1 is an enrichment marker for tumor-initiating cells in human glioblastoma. Cell Stem Cell. 2009;4(5):440–52.
Read TA et al. Identification of CD15 as a marker for tumor-propagating cells in a mouse model of medulloblastoma (vol 15, pg 135, 2009). Cancer Cell. 2009;16(3):267–7.
Suva ML et al. Reconstructing and reprogramming the tumor-propagating potential of glioblastoma stem-like cells. Cell. 2014;157(3):580–94.
Bhat KP et al. Mesenchymal differentiation mediated by NF-kappaB promotes radiation resistance in glioblastoma. Cancer Cell. 2013;24(3):331–46.
Lee J et al. Tumor stem cells derived from glioblastomas cultured in bFGF and EGF more closely mirror the phenotype and genotype of primary tumors than do serum-cultured cell lines. Cancer Cell. 2006;9(5):391–403.
Lathia JD et al. Integrin Alpha 6 regulates glioblastoma stem cells. Cell Stem Cell. 2010;6(5):421–32.
Kondo T, Setoguchi T, Taga T. Persistence of a small subpopulation of cancer stem-like cells in the C6 glioma cell line. Proc Natl Acad Sci U S A. 2004;101(3):781–6.
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Hu, J., Markowitz, G., Wang, XF. (2016). Isolation of Glioma-Initiating Cells for Biological Study. In: Koumenis, C., Coussens, L., Giaccia, A., Hammond, E. (eds) Tumor Microenvironment. Advances in Experimental Medicine and Biology, vol 899. Springer, Cham. https://doi.org/10.1007/978-3-319-26666-4_11
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DOI: https://doi.org/10.1007/978-3-319-26666-4_11
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