Analysis of the changes in the 1H NMR spectral pattern of perchloric acid extracts of C6 cells with growth.
The aim of this work was to identify spectral markers of cell proliferation that could be of use in clinical MRS. Cultured C6 ATCC rat glioma cells were used as models for this purpose and metabolites were extracted with perchloric acid at three different growth curve stages: log, confluence and post-confluence. 1D and 2D in vitro(1)H NMR spectra were recorded at 9.4 T. Statistically significant changes in myo-inositol and glutamine concentrations between log phase and post-confluence were found when normalized to the creatine ratio. The myo-inositol/creatine ratio was 2.76 +/- 0.82 at log phase increasing to 7.43 +/- 1.34 at post-confluence, while the glutamine/creatine ratio decreased from 0.22 +/- 0.03 to 0.10 +/- 0.02. No significant differences were recorded for other metabolites investigated. The fact that both myo-inositol and glutamine are detectable by in vivo MRS at clinical fields makes their changes relevant as potential astrocytic tumour proliferation rate markers in clinical MRS.
Departament de Bioquímica i Biologia Molecular, Universitat Autònoma de Barcelona, 08193 Cerdanyola del Vallès, Spain.
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D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005910:Glioma / Q000473:pathology / Q000503:physiopathology*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008657:Metabolic Clearance Rate; D010472:Perchlorates / Q000378:metabolism*; D011522:Protons; D051381:Rats; D014020:Tissue Extracts / Q000378:metabolism
D014408:Biomarkers, Tumor; D010472:Perchlorates; D011522:Protons; D014020:Tissue Extracts