Novel diagnostic and prognostic factors for the advanced melanoma based on the glycosylation-related changes studied by biophysical profiling methods

Anandamide-Modulated Changes in Metabolism, Glycosylation Profile and Migration of Metastatic Melanoma Cells

“…The GFAT upregulation also promotes cell migration and its expression correlates with poor clinical outcomes [66]. Melanoma cells undergo the epithelial-to mesenchymal-like transformation [36]; thus, for the tested cell lines in this research, it is understandable that the base level of the GFAT-1 expression for melanoma cells was significantly higher for metastatic A375-P cells than for the primary WM35 cells (approximately 138%, data available in the Supplementary Materials Table S1). At the same time, the 24 h incubation of A375-P metastatic cells with anandamide resulted in the decreased values of the relative GFAT-1 expression indicating possible inhibition of the EMT process.…”

“…This suggests a major difference in the glycosylation profile of metastatic melanoma cells treated with AEA. Although this research is the first to show the anandamide influence on cells by means of the QCM-D method, much is already known in terms of the glycosylation profile of various melanoma cells, including cell lines and primary cells isolated from a patient with confirmed melanoma metastasis [36,39,40]. With the tumor progression, the protein glycosylation on cells changes from short and lowly-branched surface glycans (such as on the WM35 cells of the primary RGP site) to long and highly branched oligosaccharides (present on the A375-P metastatic cells), thus the lectin Con A affinity for metastatic cells is higher than for the primary cells and the glycan viscoelastic index significantly increases.…”

“…With the tumor progression, the protein glycosylation on cells changes from short and lowly-branched surface glycans (such as on the WM35 cells of the primary RGP site) to long and highly branched oligosaccharides (present on the A375-P metastatic cells), thus the lectin Con A affinity for metastatic cells is higher than for the primary cells and the glycan viscoelastic index significantly increases. Because melanoma tumors are highly heterogenic, melanoma cells isolated from the same patient often differ in metastatic potential, for example, established melanoma subcultures presented a varied doubling time [75], as well as disparate cell elasticity (AFM analysis) and glycosylation profile (QCM-D analysis; [36]). However, all results were consistent: MM7 and MM9 populations were the quickest to divide (low value of doubling time), less resistant to deformation (low E values) and with high metastatic potential (low K D value and high gVI value).…”

“…Therefore, the main focus of our research was on the impact of AEA on the changes in metabolism, glycosylation profile and migration of melanoma cells from different progression stages. Due to our newly established biophysical methodology for melanoma theranostic purpose, using the quartz crystal microbalance with dissipation monitoring (QCM-D) and the direct lectin-glycan interaction approach [36], we were able to detect changes in the glycosylation profile of AEA-treated melanoma cells in real-time and right on the surface of the cells. As a model for this basic research, commercial melanoma cells derived from different progression stages (radial WM35 and vertical WM115 growth phases, lymph node WM266-4 metastasis, solid tumor A375-P metastasis) were utilized.…”

Advanced mass spectrometric and spectroscopic methods coupled with machine learning for in vitro diagnosis

“…5 The purpose of IVD is to provide information on the current or future status of the patient, often used in tasks such as risk assessment, 6 drug monitoring 7 and prognostic evaluation. 8 IVD shows great strengths in medical procession. First, IVD is usually non-or mildly-invasive, causing less harm to patients and greatly reducing their suffering compared with in vivo diagnosis.…”