1H magnetic resonance spectroscopy metabolites as biomarkers for cell cycle arrest and cell death in rat glioma cells.
BACKGROUND
Improved non-invasive imaging biomarkers of treatment response contribute to optimising cancer management and metabolites detected by proton magnetic resonance spectroscopy ((1)H MRS) show promise in this area. Understanding (1)H MRS changes occurring in cells during cell stress and cell death in vitro should aid the selection of pertinent biomarkers for clinical use.


METHODS
BT4C glioma cells in culture were exposed to either 50 μM cis-dichlorodiammineplatinum II (cisplatin) or starvation by culture in phosphate buffered saline. High resolution magic angle spinning (1)H MRS was performed on cells using a Varian 600 MHz nanoprobe and metabolites were quantified by a time domain fitting method. Cell viability was assessed by trypan blue, H&E, 4',6-diamino-2-phenylindole (DAPI), DNA laddering and annexin V-FITC labelled flow cytometry; propidium iodide flow cytometry was used to assess the cell cycle phase.


RESULTS
With cisplatin exposure, cells initially accumulated in the G1 stage of the cell cycle with low numbers of apoptotic and necrotic cells and this was associated with decreases in phosphocholine, succinate, alanine, taurine, glycine and glutamate and increases in lactate and glycerophosphocholine (GPC). Starvation, leading to necrotic cell death within 6-18 h, caused decreases in succinate, alanine, glycine, and glutamate and increases in GPC. Principal component analysis revealed two patterns of metabolite changes, one common to both types of cell stress and another specific for necrosis secondary to cell starvation.


CONCLUSIONS
(1)H MRS reveals alterations in multiple metabolites during cell cycle arrest and cell death which may provide early biomarker profiles of treatment efficacy in vivo.
Mirbahai
Ladan
L
School of Sport and Exercise Sciences, University of Birmingham, Edgbaston, Birmingham, UK.
Wilson
Martin
M
Shaw
Christopher S
CS
McConville
Carmel
C
Malcomson
Roger D G
RD
Griffin
Julian L
JL
Kauppinen
Risto A
RA
Peet
Andrew C
AC
D000596:Amino Acids / Q000032:analysis / Q000378:metabolism; D000818:Animals; D014408:Biomarkers, Tumor / Q000032:analysis / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects*; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000627:therapeutic use*; D016193:G1 Phase / Q000187:drug effects*; D005910:Glioma / Q000378:metabolism / Q000628:therapy; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D010767:Phosphorylcholine / Q000032:analysis / Q000378:metabolism; D011522:Protons; D051381:Rats; D019802:Succinic Acid / Q000032:analysis / Q000378:metabolism
D000596:Amino Acids; D014408:Biomarkers, Tumor; D011522:Protons; D010767:Phosphorylcholine; D019802:Succinic Acid; D002945:Cisplatin