A HIF-independent, CD133-mediated mechanism of cisplatin resistance in glioblastoma cells

Cellular Oncology (Dordrecht)
Eroje M AhmedAnna Grabowska

Abstract

Glioblastoma (GBM) is the commonest brain tumour in adults. A sub-population of cells within these tumours, known as cancer stem cells (CSCs), is thought to mediate their chemo-/radiotherapy resistance. CD133 is a cell surface marker that is used to identify and isolate GBM CSCs. However, its functional significance, as well as the relevant microenvironment in which to study CD133, have so far remained unknown. Here, we examined the effect of hypoxia on the expression of CD133 and on that of the hypoxia-related factors HIF-1α and HIF-2α, and the potential functional significance of CD133 expression on the acquisition of chemo-resistance by GBM cells. CD133, HIF-1α, HIF-2α, VEFG and (control) HPRT mRNA expression analyses were carried out on GBM cells (U251, U87 and SNB19; 2D or 3D cultures) under both normoxic and hypoxic conditions, using qRT-PCR. siRNA was used to downregulate CD133, HIF-1α and HIF-2α expression in the GBM cells, which was confirmed by flow cytometry and qRT-PCR, respectively. Drug sensitivity-related IC50 values were established using an Alamar Blue cell viability assay in conjunction with the Graphpad prism software tool. We found that the expression of CD133 was upregulated under hypoxic conditions in both...Continue Reading

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Citations

Oct 12, 2018·International Journal of Molecular Sciences·Stéphane TerrySalem Chouaib
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Methods Mentioned

BETA
flow cytometry
transfection

Software Mentioned

Weasel
Graphpad prism

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