PLEKHG5 regulates autophagy, survival and MGMT expression in U251-MG glioblastoma cells
Witte KE, Slotta C, Luetkemeyer M, Kitke A, Coras R, Simon M, Kaltschmidt C, Kaltschmidt B (2020)
Publication Type: Journal article
Publication year: 2020
Journal
Book Volume: 10
Journal Issue: 1
DOI: 10.1038/s41598-020-77958-3
Abstract
A signalling pathway involving PLEKHG5 (guanine exchange factor) for the Ras superfamily member RAB26 to transcription factor NF-kappa B was discovered in autophagy. PLEKHG5 was reported in glioblastoma multiforme (GBM) and correlates with patient survival. Thus, the generation of a cellular model for understanding PLEKHG5 signalling is the study purpose. We generated a CRISPR/Cas9-mediated knockout of PLEKHG5 in U251-MG glioblastoma cells and analysed resulting changes. Next, we used a mRFP-GFP-LC3(+) reporter for visualisation of autophagic defects and rescued the phenotype of PLEKHG5 wildtype via transduction of a constitutively active RAB26QL-plasmid. Effects of overexpressing RAB26 were investigated and correlated with the O-6-methylguanine-DNA methyltransferase (MGMT) and cellular survival. PLEKHG5 knockout showed changes in morphology, loss of filopodia and higher population doubling times. Accumulation of autolysosomes was resulted by decreased LAMP-1 in PLEKHG5-deficient cells. Rescue of PLEKHG5(-/-) restored the downregulation of RhoA activity, showed faster response to tumour necrosis factor and better cellular fitness. MGMT expression was activated after RAB26 overexpression compared to non-transduced cells. Survival of PLEKHG5 knockout was rescued together with sensitivity to temozolomide by RAB26QL. This study provides new insights in the PLEKHG5/RAB26 signalling within U251-MG cells, which suggests potential therapeutic strategies in other glioma cells and further in primary GBM.
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Germany (DE)How to cite
APA:
Witte, K.E., Slotta, C., Luetkemeyer, M., Kitke, A., Coras, R., Simon, M.,... Kaltschmidt, B. (2020). PLEKHG5 regulates autophagy, survival and MGMT expression in U251-MG glioblastoma cells. Scientific Reports, 10(1). https://doi.org/10.1038/s41598-020-77958-3
MLA:
Witte, Kaya Elisa, et al. "PLEKHG5 regulates autophagy, survival and MGMT expression in U251-MG glioblastoma cells." Scientific Reports 10.1 (2020).
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