Metabolic Markers of MG-63 Osteosarcoma Cell Line Response to Doxorubicin and Methotrexate Treatment: Comparison to Cisplatin

Abstract

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 UDPGlcNAc 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.

Chemotherapeutic response to cisplatin-like drugs in human breast cancer cells probed by vibrational microspectroscopy

“…Following previous biological studies by the authors on the effect of the tested Pt(II) and Pd(II) spermine complexes on the growth and viability of several human cancer cells, [14][15][16]22,26,27,40,41,43,45 including the hormone-unresponsive breast carcinoma currently investigated, a cellular response at the biochemical level is presently sought, through analysis of the vibrational pattern of drug-treated and untreated cells. This multidisciplinary approach aims to link biological response to changes in cellular chemistry and metabolism, thus identifying the spectral markers of drug activity and paving the way for an understanding of the drug mode of action at the molecular level, hopefully helping to predict drug efficacy under specic conditions.…”

“…The results obtained along this work were related with a thorough conformational analysis of the newly synthesised metal-based agents (by Raman, IR and INS coupled to theoretical approaches) [18][19][20][21][22][23][24][34][35][36] and with biological assays for the evaluation of cytotoxicity performed alongside on the same systems -cell growth and viability inhibition, as well as anti-angiogenic and anti-metastatic abilities [14][15][16]27,[40][41][42][43]45 -allowing us to establish a reliable correlation between structural/ conformational features, cytotoxic activity and impact on cellular metabolism. This data, coupled to previous INS and EXAFS studies on drug-DNA adducts (carried out at ISIS and DLS 18,21,23,24 ), will pave the way for future work on other promising cisplatin-like anticancer agents (both Pt(II) and Pd(II)) which have been studied by the team in the last few years, as well as on distinct human cancer cells.…”

“…The data obtained within this work, coupled to conformational analysis of the metal complexes and cytological assays (for evaluation of growth-inhibition, cytotoxic and anti-metastatic activities) previously carried out at QFM-UC, [14][15][16]22,[25][26][27][40][41][42][43][44][45] will pave the way for future studies on other platinum and palladium-based polyamine agents prepared by the authors. This approach should provide valuable clues for the rational design of novel cisplatin-like drugs, displaying a higher efficiency, lower toxicity, decreased acquired resistance and possible oral administration.…”

“…A thorough knowledge of these compounds and of their interactions with their main pharmacological target (DNA), as well as with other biomolecules involved in the drug pharmacokinetics (e.g. glutathione) and pharmacodynamics, has been attained through vibrational spectroscopy (Raman, FTIR and Inelastic Neutron Scattering (INS)) [18][19][20][21][22][23][24] coupled to synchrotron-based Extended X-ray Absorption Fine Structure (EXAFS), 23 as well as by NMR metabolomics 22,25 and electrochemical techniques. 26,27 In order to design such effective chemotherapeutic agents, it is essential not only to determine their effect on cellular growth, metabolism and function, but also to understand the mechanisms (at a molecular level) associated with this biological impact.…”

Application of metabolomics in sarcoma: From biomarkers to therapeutic targets

“…Methods based on NMR can measure the liquid and solid samples at once with a minimal preparation stage. High-resolution magic angel spinning NMR (HR-MAS-NMR) spectroscopy is widely tested for metabolite-based sarcoma classification [33, 34]. More recently, direct analysis of spatial distribution of metabolites in a tissue is an emerging approach in metabolomics.…”