resumo
A high resolution magic angle spinning NMR metabolomics study of the effects of doxorubicin (DOX), methotrexate (MTX) and cisplatin (cDDP) on MG-63 cells is presented and unveils the cellular metabolic adaptations to these drugs, often used together in clinical protocols. Although cDDP-treated cells were confirmed to undergo extensive membrane degradation accompanied by increased neutral lipids, DOX- and MTX-treated cells showed no lipids increase and different phospholipid signatures, which suggests that (i) DOX induces significant membrane degradation, decreased membrane synthesis, and apparent inhibition of de novo lipid synthesis, and (ii) MTX induces decreased membrane synthesis, while no membrane disruption or de novo lipid synthesis seem to occur. Nucleotide signatures were in apparent agreement with the different drug action mechanisms, a link having been found between UDP-GlcNAc and the active pathways of membrane degradation and energy metabolism, for cDDP and DOX, with a relation to oxidative state and DNA degradation, for cDDP. Correlation studies unveiled drug-specific antioxidative signatures, which pinpointed m- and s-inositols, taurine, glutamate/glutamine, and possibly creatine as important in glutathione metabolism. These results illustrate the ability of NMR metabolomics to measure cellular responses to different drugs, a first step toward understanding drug synergism and the definition of new biomarkers of drug efficacy.
palavras-chave
MAGNETIC-RESONANCE-SPECTROSCOPY; OF-THE-ART; K562 CELLS; MASS-SPECTROMETRY; INDUCED APOPTOSIS; VISIBLE LIPIDS; LEUKEMIA-CELLS; CANCER CELLS; TUMOR-CELLS; H-1-NMR
categoria
Biochemistry & Molecular Biology
autores
Lamego, I; Duarte, IF; Marques, MPM; Gil, AM
nossos autores
Ana Maria Pissarra Coelho Gil
Professor Associado com Agregação
Inês Dias Lamego
Estudante de doutoramento
Iola Melissa Fernandes Duarte
Investigador Principalagradecimentos
The authors acknowledge funding from the European Regional Development Fund-FEDER through the Competitive Factors Thematic Operational Programme-COMPETE and the Foundation for Science and Technology-FCT, Portugal (CICECOFCOMP-01-0124-FEDER-037271, PEst-OE/QUI/UI0070 / 2014, PEst-C/CTM/LA0011/2013, PTDC/5AL-BEB-66896/ 2006, and SFRH/BD/63916/2009). The Portuguese National NMR Network (RNRNIN), supported with FCT funds, is acknowledged (particularly for access to 800 MHz NMR equipment). The authors thank M. Spraul, Bruker BioSpin, Germany, for access to software and spectral databases, and the Associate Laboratory IBMC-INEB for kindly providing the MG63 cell line.