AbstractConstitutive activation of the Wnt pathway plays a key role in the development of colorectal cancer and has also been implicated in the pathogenesis of other malignancies. Deregulation of Wnt signaling mainly occurs through genetic alterations of APC, the β‐catenin gene (CTNNB1), AXIN1 and AXIN2, leading to stabilization of β‐catenin. Physiologically, AXIN2 is transcriptionally induced on Wnt signaling activation and acts as a negative feedback regulator of the pathway. In colorectal cancer, mutations in CTNNB1 and AXIN2 occur preferentially in tumors with inactivation of the mismatch repair (MMR) genes MSH2, MLH1, or PMS2. In this study, the expression of β‐catenin and AXIN2, and the mutational status of CTNNB1, APC, and AXIN2 were evaluated in two MMR‐deficient (PR‐Mel and MR‐Mel) and seven MMR‐proficient human melanoma cell lines. Only PR‐Mel and MR‐Mel cells showed nuclear accumulation of β‐catenin and expression of the AXIN2 gene, and hence, constitutive activation of Wnt signaling. Mutational analysis identified a somatic heterozygous missense mutation in CTNNB1 exon three and a germline heterozygous deletion within AXIN2 exon seven in PR‐Mel cells, and a somatic biallelic deletion within APC in MR‐Mel cells. Deregulation of Wnt signaling and a defective MMR system were also present in the original tumor of PR and MR patients. Thus, we describe additional melanomas with mutations in CTNNB1 and APC, identify for the first time a germline AXIN2 mutation in a melanoma patient and suggest that inactivation of the MMR system and deregulation of the Wnt/β‐catenin signaling pathway cooperate to promote melanoma development and/or progression. © 2008 Wiley‐Liss, Inc.