While searching a causal association between anoikis resistance and malignant transformation, we developed a melanomagenesis model, where different melanoma cell lines (4C3, 4C11, Tm1, Tm5 and others) were established after submitting a non-tumorigenic murine melanocyte lineage (melan-a) to five sequential substrate adhesion blockade. Non-tumorigenic melan-a-derived cell sublines (1C, 2C, 3C and 4C), presenting increasing anoikis resistance and altered expression of several genes (including mdm2 and p53), were also obtained after each adhesion impediment cycle. A marked increment in global DNA methylation was observed in melan-a cells maintained in suspension and a progressive global DNA hypomethylation in melan-a-derived cell lines (2C, 4C, 4C3, 4C11, Tm1 and Tm5). Furthermore, DNA methyltransferases (DNMTs) expression modifies significantly throughout melan-a transformation. An increase in DNMT3b expression is observed in melan-a cells maintained in suspension, while melan-a-derived cell lines do not express this protein anymore. In addition, DNA methyltransferase 3a transcripts are augmented in melanoma cell lines. DNA demethylation of Tm5 melanoma cells by 5’-Aza-2’-deoxycytidine resulted in a drastic reduction in their in vivo tumor growth, indicating the involvement of DNA methylation in carcinogenesis associated with substrate adhesion deregulation. Until now, at least two genes whose expression seems to be regulated by DNA methylation were already identified in this model: p19^ARF tumor supressor, involved in cell cycle control; and TIMP1, whose expression was recently related to anoikis resistance in tumor cells. These data show that a perturbation of methylation profile occurs along the transformation process, which is already detectable few hours after melan-a deadhesion. The presence of non-tumorigenic cells in a new microenvironment might result in altered methylation status, leading to modified expression of some genes and contributing to the acquisition of a gene expression profile favorable to cell transformation.