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

Abstract

Melanoma is the most aggressive type of skin cancer, leading to metabolic rewiring and enhancement of metastatic transformation. Efforts to improve its early and accurate diagnosis are largely based on preclinical models and especially cell lines. Hence, we herein present a combinational Nuclear Magnetic Resonance (NMR)- and Ultra High Performance Liquid Chromatography-High-Resolution Tandem Mass Spectrometry (UHPLC-HRMS/MS)-mediated untargeted metabolomic profiling of melanoma cells, to landscape metabolic alterations likely controlling metastasis. The cell lines WM115 and WM2664, which belong to the same patient, were examined, with WM115 being derived from a primary, pre-metastatic, tumor and WM2664 clonally expanded from lymph-node metastases. Metabolite samples were analyzed using NMR and UHPLC-HRMS. Multivariate statistical analysis of high resolution NMR and MS (positive and negative ionization) results was performed by Principal Component Analysis (PCA), Partial Least Squares-Discriminant Analysis (PLS-DA) and Orthogonal Partial Least Squares-Discriminant Analysis (OPLS-DA), while metastasis-related biomarkers were determined on the basis of VIP lists, S-plots and Student’s t-tests. Receiver Operating Characteristic (ROC) curves of NMR and MS data revealed significantly differentiated metabolite profiles for each cell line, with WM115 being mainly characterized by upregulated levels of phosphocholine, choline, guanosine and inosine. Interestingly, WM2664 showed notably increased contents of hypoxanthine, myo-inositol, glutamic acid, organic acids, purines, pyrimidines, AMP, ADP, ATP and UDP(s), thus indicating the critical roles of purine, pyrimidine and amino acid metabolism during human melanoma metastasis.

Genetic Target Modulation Employing CRISPR/Cas9 Identifies Glyoxalase 1 as a Novel Molecular Determinant of Invasion and Metastasis in A375 Human Malignant Melanoma Cells In Vitro and In Vivo

“…Metabolic reprogramming is a molecular hallmark of cancer that has also been substantiated in melanoma. However, even though GLO1 expression serves an essential function in the maintenance of glycolytic energy metabolism, the specific role of GLO1 in melanomagenesis remains largely unexplored [ 2 , 5 , 22 , 38 , 39 ].…”

From Proteomic Mapping to Invasion-Metastasis-Cascade Systemic Biomarkering and Targeted Drugging of Mutant BRAF-Dependent Human Cutaneous Melanomagenesis

“…Metabolic re-programming is tightly associated with the tumor’s metastatic potential. Since the MCT1 lactate transporter (importer) plays diverse critical roles in the metastasis process [ 218 , 219 , 220 , 221 , 222 , 223 ], we next analyzed the cellular localization and distribution pattern of the MCT1 ( SLC16A1 /MOT1) transporter in WM115 and WM266-4 (2.70) ( Table S3 , [ 35 ]) melanoma cells ( Figure 7 C). Surprisingly, through immunofluorescence imaging, MCT1 was presented to strongly accumulate in the melanoma nucleus and not the cell membrane (as expected) for both cell types ( Figure 7 C).…”

Inhibitory Effect of Dihydroartemisinin on the Proliferation and Migration of Melanoma Cells and Experimental Lung Metastasis From Melanoma in Mice

“…Recent studies have demonstrated that metabolism is significantly associated with tumorigenesis and metastasis ( Luengo et al, 2017 ; Kosmopoulou et al, 2020 ; Li et al, 2016 ). Based on the strategy of tumor immunotherapy with metabolic regulation, UHPLC-Q-TOF-MS technology was used.…”