NMR metabolic fingerprints of murine melanocyte and melanoma cell lines: application to biomarker discovery

Abstract

Melanoma is the most aggressive type of skin cancer and efforts to improve the diagnosis of this neoplasia are largely based on the use of cell lines. Metabolomics is currently undergoing great advancements towards its use to screening for disease biomarkers. Although NMR metabolomics includes both 1D and 2D methodologies, there is a lack of data in the literature regarding heteronuclear 2D NMR assignments of the metabolome from eukaryotic cell lines. The present study applied NMR-based metabolomics strategies to characterize aqueous and lipid extracts from murine melanocytes and melanoma cell lines with distinct tumorigenic potential, successfully obtaining fingerprints of the metabolites from the extracts of the cell lines by means of 2D NMR HSQC correlation maps. Relative amounts of the identified metabolites were compared between the 4 cell lines. Multivariate analysis of 1H NMR data was able not only to differentiate the melanocyte cell line from the tumorigenic ones but also distinguish among the 3 tumorigenic cell lines. We also investigated the effects of mitogenic agents, and found that they can markedly influence the metabolome of the melanocyte cell line, resembling the pattern of most proliferative cell lines.

Human Melanoma-Cell Metabolic Profiling: Identification of Novel Biomarkers Indicating Metastasis

“…The elevated concentration of creatine (FC NMR = 4.91) and phosphocreatine (FC NMR = 2.53) in WM2664 cells mainly reflects their essential roles in energy storage required under the increased metabolic rates of tumor cells. In addition to synthesis of ATP, these two metabolites are also involved in its trafficking from the internal mitochondrial membrane to the cytoplasm, thus acting as energy distribution regulators [38,39]. However, ATP, the most important molecule in the cell-energy network and homeostasis, is predominantly formed during the glycolysis process, as well as by oxidative phosphorylation in mitochondria.…”

NMR Spectroscopy for Metabolomics Research

“…NMR spectroscopy plays important and multifaceted roles that have benefited and continue to benefit the field of metabolomics. These include biomarker discovery [51,259,260,261,262], kinetic and mechanistic metabolic studies [263,264,265,266,267], metabolite imaging [268,269,270], food and beverage characterization [271,272,273,274], environmental monitoring [275,276,277], and drug toxicity studies [278,279,280,281,282,283,284]. The unique characteristics of NMR, including its high level of reproducibility, its simplicity in sample preparation, its capacity to handle diverse sample types (liquids, solids, gels), its quantitative capabilities, and its utility in identifying unknown metabolites along with its nondestructive nature, have made it particularly useful in many diverse metabolomic disciplines.…”

Discovery of potential biomarkers in human melanoma cells with different metastatic potential by metabolic and lipidomic profiling

“…Comparative metabolite profiling between melanoma and non-melanoma lesions in patients, and profiling of metabolic fluxes of human melanoma cells and melanocytes using gas chromatography-mass spectrometry (GC-MS) have been reported 16 , 17 . In addition, metabolic changes related to metastasis of melanoma in a C57BL/6J mouse model and murine melanoma cell lines (Tm1, Tm5, and B16F10) have been identified by nuclear magnetic resonance (NMR)-based metabolic profiling 18 – 21 , and a fatty acids and phospholipid group analysis of high- and low-metastatic B16 melanoma cell lines by gas-liquid chromatography equipped with dual-flame ionization detectors has been performed 22 . However, there has been no research focused on comparing changes in metabolites and individual lipid species in human melanoma cell lines with low and high metastatic potential with mass spectrometric-based approaches.…”