IDH1 R132H gene mutation reduces cell proliferation and sensitizes recurrent Glioblastoma to hydrogen peroxide

Abstract

Glioblastoma (GBM) recurrence rate is 90% resulting in 15 months median survival only. Isocitrate dehydrogenase 1 (IDH1) mutations in gliomas significantly improved patient’s prognosis. Therefore, understanding common IDH1 mutation, IDH1 R132H in recurrent GBM is necessary to improve poor survival rate. IDH1 R132H recurrent GBM was developed to investigate cell proliferation rate and sensitivity towards oxidative stress induced by hydrogen peroxide. The cell death mechanism induced by hydrogen peroxide were further investigated. Malaysian recurrent GBM cell line was authenticated via Short Tandem Repeat and screened for IDH1 gene via PCR. IDH1 R132H gene expression in GBM was confirmed via real-time PCR and western blot. The effect of IDH1 R132H mutation on cell proliferation rate and cytotoxicity using hydrogen peroxide were determined using MTT assay. The angiogenesis, apoptotic genes and cell cycle induced by hydrogen peroxide in IDH1 wild-type GBM were determined via real-time PCR and flow-cytometry. Malaysian GBM cell line is unique and harbors IDH1 wild-type gene. IDH1 R132H gene mutation significantly reduced the growth rate and sensitized the GBM cells to hydrogen peroxide at 72 hours (p<0.05). Hydrogen peroxide induced significant G1 cell cycle arrest and apoptosis in IDH1 wild-type GBM cell line (p<0.05). Slower growth rate and higher sensitivity towards oxidative stress may explain why IDH1 mutant patients have better prognosis compared to IDH1 wild-type patients. Confirmation GBM cell death mechanism via hydrogen peroxide showed that it has potential to treat gliomas.

Keywords

• Glioblastoma,IDH1 R132H,Proliferation rate,Cell cycle,Oxidative stress

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