Molecular subtyping of metastatic melanoma based on cell ganglioside metabolism profilesReport as inadecuate

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BMC Cancer

, 14:560

Translational oncology


BackgroundIn addition to alterations concerning the expression of oncogenes and onco-suppressors, melanoma is characterized by the presence of distinctive gangliosides sialic acid carrying glycosphingolipids. Gangliosides strongly control cell surface dynamics and signaling; therefore, it could be assumed that these alterations are linked to modifications of cell behavior acquired by the tumor. On these bases, this work investigated the correlations between melanoma cell ganglioside metabolism profiles and the biological features of the tumor and the survival of patients.

MethodsMelanoma cell lines were established from surgical specimens of AJCC stage III and IV melanoma patients. Sphingolipid analysis was carried out on melanoma cell lines and melanocytes through cell metabolic labeling employing 3-Hsphingosine and by FACS. N-glycolyl GM3 was identified employing the 14 F7 antibody. Gene expression was assayed by Real Time PCR. Cell invasiveness was assayed through a Matrigel invasion assay; cell proliferation was determined through the soft agar assay, MTT, and H thymidine incorporation. Statistical analysis was performed using XLSTAT software for melanoma hierarchical clustering based on ganglioside profile, the Kaplan-Meier method, the log-rank Mantel-Cox test, and the Mantel-Haenszel test for survival analysis.

ResultsBased on the ganglioside profiles, through a hierarchical clustering, we classified melanoma cells isolated from patients into three clusters: 1 cluster 1, characterized by high content of GM3, mainly in the form of N-glycolyl GM3, and GD3; 2 cluster 2, characterized by the appearance of complex gangliosides and by a low content of GM3; 3 cluster 3, which showed an intermediate phenotype between cluster 1 and cluster 3. Moreover, our data demonstrated that: a a correlation could be traced between patients’ survival and clusters based on ganglioside profiles, with cluster 1 showing the worst survival; b the expression of several enzymes sialidase NEU3, GM2 and GM1 synthases involved in ganglioside metabolism was associated with patients’ survival; c melanoma clusters showed different malignant features such as growth in soft agar, invasiveness, expression of anti-apoptotic proteins.

ConclusionsGanglioside profile and metabolism is strictly interconnected with melanoma aggressiveness. Therefore, the profiling of melanoma gangliosides and enzymes involved in their metabolism could represent a useful prognostic and diagnostic tool.

KeywordsGanglioside Melanoma N glycolyl GM3 Sialidase Survival AbbreviationsAJCCAmerican Joint Committee on Cancer

d-GM3de-N-acetyl GM3

Neu5Gc-GM3N glycolyl GM3

Neu5Ac-GM3N acetyl GM3

VPGVertical growth phase

NHEM-AdAdult melanocytes

NHEM-NeoNeonatal melanocytes


CMAHCMP-NeuAc hydroxylase

GM3 synthaseST3 beta-galactoside-alpha-2,3-sialyltransferase-5

GD3 synthaseST8 alpha-N-acetyl-neuraminide-alpha-2-8-sialyltransferase

GM2 synthaseBeta-1,4-N-acetyl-galactosaminyltransferase-1

GM1 synthaseUDP-gal,betaGlcNAc-beta-1,3-galactosyltransferase

GD1a synthaseST6 alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3-N-acetylgalactosaminide alpha-2,6-sialyltransferase.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2407-14-560 contains supplementary material, which is available to authorized users.

Cristina Tringali, Ilaria Silvestri contributed equally to this work.

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Author: Cristina Tringali - Ilaria Silvestri - Francesca Testa - Paola Baldassari - Luigi Anastasia - Roberta Mortarini - Andrea An


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