Gene expression profiling of primary cutaneous melanoma.

Cutaneous Malignant Melanoma (CMM) is the most malignant skin tumour in humans, the incidence of which is rising rapidly in most fair-skinned populations, without apparent decline in mortality. Both hereditary, constitutional and environmental factors play a role in its etiology. CMM arises from melanocytes in the epidermis, and proceeds through discrete steps of tumor-progression that consist histologically of the radial growth phase (RGP), vertical growth phase (VGP) and metastatic phase. The underlying molecular mechanisms that govern the transition between these growth phases are hardly known. The prognosis of patients with VGP melanoma depends on several clinical and histological parameters; the latter include thickness, mitotic activity, presence or absence of ulceration and regression, and pattern of lymphocytic host response. However, there is still need for new prognostic parameters. To obtain insight in the molecular mechanisms of tumor progression in CMM, and in search of new prognostic markers, we performed global gene-expression profiling using 44K oligonucleotide micro-arrays on a unique retrospective series of 83 frozen primary MM with VGP, 9 metastases and 23 benign nevi. Unsupervised analysis allowed us o identify clusters of melanoma patients with different outcome based on their gene expression profile only. Supervised analysis resulted in the identification of a genomic signature of 254 genes with prognostic significance. The large majority of the 254 enes was correlated with thickness, thereby stressing the importance of thickness in he prognosis of CMM. This signature was validated on a separate series of melanoma patients, and proved to have a predictive accuracy comparable to what can be obtained by tumour thickness and ulceration. On an immunohistochemical level, we identified 8 new markers that may help in the prognostication of melanoma patients; three of these markers, i.e. the mini-chromosome maintenance (mcm) proteins mcm3, 4 and 6, hat are involved in DNA-replication, had independent prognostic value. Additionally, upervised analysis showed similarities in gene expression profile between primary CMM and their metastases. In conclusion, our data provide new information regarding the molecules that are operative in the progression of CMM. CMM is notorious for its resistance to chemotherapy, and disseminated CMM is a uniformly fatal disease. As several of the progression-related genes, encode molecules that have been the target of established or xperimental cancer therapies, our results may hopefully contribute to the treatment of end-stage CMM-patients.