title: Lipid biomarkers of glioma cell growth arrest and cell death detected by 1 H magic angle spinning MRS.
abstract: Biomarkers of early response to treatment have the potential to improve cancer therapy by allowing treatment to be tailored to the individual. Alterations in lipids detected by in vivo MRS have been suggested as noninvasive biomarkers of cell stress and early indicators of cell death. An improved understanding of the relationship between MRS lipids and cell stress in vitro would aid in the translation of this technique into clinical use. Rat BT4C glioma cells were treated with 50 µ m cis-dichlorodiammineplatinum II (cisplatin), a commonly used chemotherapeutic agent, and harvested at several time points up to 72 h. High-resolution magic angle spinning (1) H MRS of cells was then performed on a 600-MHz NMR spectrometer. The metabolites were quantified using a time domain fitting method, TARQUIN. Increases were detected in saturated and polyunsaturated fatty acid resonances early during the exposure to cisplatin. The fatty acid CH(2) /CH(3) ratio was unaltered by treatment after allowing for contributions of macromolecules. Polyunsaturated fatty acids increased on treatment, with the group -CH=CH-CH(2) -CH=CH- accounting for all the unsaturated fatty acid signals. Transmission electron microscopy, in addition to Nile red and 4',6-diamino-2-phenylindole co-staining, revealed that the lipid increase was associated with cytoplasmic neutral lipid droplets. Small numbers of apoptotic and necrotic cells were detected by trypan blue, annexin V-fluorescein isothiocyanate-labelled flow cytometry and DNA laddering after up to 48 h of cisplatin exposure. Propidium iodide flow cytometry revealed that cells accumulated in the G1 stage of the cell growth cycle. In conclusion, an increase in the size of the lipid droplets is detected in morphologically viable cells during cisplatin exposure. (1) H MRS can detect lipid alterations during cell cycle arrest and progression of cell death, and has the potential to provide a noninvasive biomarker of treatment efficacy in vivo.
authors.affiliation: School of Sport and Exercise Sciences, University of Birmingham, Edgbaston, Birmingham, UK. ladan.mirbahai@bindingsite.co.uk
authors.forename: Ladan
authors.initials: L
authors.lastname: Mirbahai
authors.forename: Martin
authors.initials: M
authors.lastname: Wilson
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Shaw
authors.forename: Carmel
authors.initials: C
authors.lastname: McConville
authors.forename: Roger D G
authors.initials: RD
authors.lastname: Malcomson
authors.forename: Risto A
authors.initials: RA
authors.lastname: Kauppinen
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Peet
mesh_terms: D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000378:metabolism*; D001932:Brain Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D002453:Cell Cycle / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology; D053938:DNA Fragmentation / Q000187:drug effects; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005910:Glioma / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D007211:Indoles / Q000378:metabolism; D008055:Lipids / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D010078:Oxazines / Q000378:metabolism; D011419:Propidium / Q000378:metabolism; D011522:Protons; D051381:Rats; D013194:Staining and Labeling; D014343:Trypan Blue / Q000378:metabolism
chemical_list: D017304:Annexin A5; D014408:Biomarkers, Tumor; D007211:Indoles; D008055:Lipids; D010078:Oxazines; D011522:Protons; D011419:Propidium; C007293:DAPI; D016650:Fluorescein-5-isothiocyanate; D014343:Trypan Blue; C044808:nile red; D002945:Cisplatin