Background

Genes of the Raf family encode kinases that are regulated by Ras and mediate cellular responses to growth signals. Recently, it was shown that activating mutations of BRaf are found with high frequency in human melanomas. The Ras family member most often mutated in melanoma is NRas.

Methods

The constitutive activation of the Ras/Raf signaling pathway suggests an impact on the clinical course of the tumor. To address this notion, we analyzed tumor DNA from 114 primary cutaneous melanomas and of 86 metastatic lesions obtained from 174 patients for mutations in BRaf (exons 15 and 11) and NRas (exons 1 and 2) by direct sequencing of PCR products and correlated these results with the clinical course.

Results

In 57.5% of the tumors either BRaf or NRas were mutated with a higher incidence in metastatic (66.3%) than in primary lesions (50.9%). Although the majority of BRaf mutations affected codon 599, almost 15% of mutations at this position were different from the well-described exchange from valine to glutamic acid. These mutations (V599R and V599K) also displayed increased kinase and transforming activity. Surprisingly, the additional BRaf variants D593V, G465R and G465E showed a complete loss of activity in the in vitro kinase assay; however, cells overexpressing these mutants displayed increased Erk phosphorylation. The correlation of mutational status and clinical course revealed that the presence of BRaf/NRas mutations in primary tumors did not negatively impact progression free or overall survival. In contrast, however, for metastatic lesions the presence of BRAF/NRAS mutations was associated with a significantly poorer prognosis, ie a shortened survival.

Conclusion

We demonstrate a high–albeit lower than initially anticipated–frequency of activating BRaf mutations in melanoma in the largest series of directly analyzed tumors reported to date. Notably, the clinical course of patients harboring activating BRaf mutations in metastatic melanoma was significantly affected by the presence of a constitutive BRaf activation in these.

Background

There are about 133,000 new cases of melanoma worldwide each year, of which almost 80% occur in the Caucasian population of North America, Europe, Australia and New Zealand [1]. The clinical features of melanoma are asymmetry, a coastline border, or multiple colors, ie signs reflecting the heterogeneity of the melanoma cells within one tumor. The classification system supersedes histopathological parameters and clinical features, thereby superficial spreading melanoma (SSM), nodular melanoma (NM), acral-lentiginous melanoma (ALM), and lentigo maligna melanoma (LMM) are differentiated. However, prognosis of primary melanoma is essentially determined by vertical tumor thickness and the presence or absence of ulceration [2].