Biomarkers of cell stress and cell death detected by proton high resolution magic angle spinning (\(^1\)H HR-MAS) nuclear magnetic resonance (NMR) spectroscopy in a rat glioma cell line

Abstract

Early detection of biomarkers of tumour treatment response improves clinical management, in vivo. Magnetic resonance spectroscopy (MRS) has demonstrated potential for identifying early biomarkers of effective treatment. However, more detailed in vitro studies are required to improve our understanding and facilitate its use. The aim of this study is to determine \(^1\)H high-resolution magic angle spinning (HR-MAS) nuclear magnetic resonance (NMR) biomarkers of cytostasis and cell death in a rat glioma BT4C cell line. Cytostasis and cell death were induced in BT4C cells using cisplatin and substrate free medium, respectively. Cell viability was examined by various techniques. The lipid and metabolite alterations in whole cells were investigated by \(^1\)H HR-MAS NMR. Significant alterations in lipids and metabolites were detected in response to cytostasis or necrosis. NMR lipid accumulation was associated with an increase in cytoplasmic lipid droplets seen prior to morphological and molecular markers of cell death. Significant differences were detected in individual choline containing metabolites (CCMs), emphasising the importance of identifying CCMs separately. Alterations were also detected in lactate, alanine, glycine, glutamate, and succinate levels, suggesting changes in the energy metabolism pathways which may provide novel biomarkers in vivo. \(^1\)H HR-MAS NMR reveals alterations in lipids and metabolites during cytostasis and cell death which may provide early markers of treatment efficacy.