1 Nima Rezvani Kakhki, Department of pharmaceutical sciences, Budapest
2 Marta Kraszni Mazakne, Department of pharmaceutical sciences, Budapest
3 Bela Noszal, Department of pharmaceutical sciences, Budapest
90% of skin cancer mortality is caused by melanoma, a malignant symptom, infamous for its ability to metastasize. Changes in the metabolome are molecular consequences from perturbations in cellular activity. Metabolic profiling is therefore a promising tool for early recognition, and the development of its specific therapy, an unmet medical need.
The aim of our research was to find distinctive metabolic properties between melanoma cell lines with low (WM983B) and high (HT168M1) metastatic potential using high field Nuclear Magnetic Resonance (NMR) spectroscopy combined with chemometric statistical analysis to assign the hundreds of 1H NMR peaks.
Extraction of the polar metabolites from the cultured cells was performed with a methanol/chloroform/water 10/10/9 mixture. The aqueous/methanolic phase was separated and dried under a nitrogen gas stream. All dried extracts were re-dissolved in D2O containing TSP reference, and the pH was set to 7.00 +/-0.02. 600 MHz NMR spectra of the solutions were recorded and uniformly processed. Data matrices were built from the integrals of the NMR spectra using 0.02 ppm bins and introduced to the Metaboanalyst program for statistical analysis.
Partial least squares projection to latent structure discriminant analysis (PLS-DA) of the data showed clear differences among the cell lines investigated. The most important, distinctive metabolites were glycerophosphocholine (GPC), gluthathione, lactic acid and myoinositol showing highest level in the highly metastatic cell line and lowest level in the healthy one. In addition there is less, but still significant difference in the alanine, taurine and leucine levels. These changes in the small molecule metabolic profile facilitate the proliferation of cancerous cells and provide potential biomarkers to differentiate between melanoma cells with different metastatic potential.
Doctoral School: Pharmaceutical Sciences
Program: Modern Trends in Pharmaceutical Scientific Research
Supervisor: Bela Noszal, Marta Kraszni Mazakne
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