Molecular cloning of genes expressed downstream the p51 pathway in melanocytes and keratinocytes

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One of the p53 family members, p51(p63,p73L or Ket) has been reported to be essential for formation of cutaneous organ and adnexal tissues. By using recombinant adenoviruses that express one of the p53 family proteins, suppression of growth and induction of apoptosis were studied in melanoma cell lines. We found that among p53 family proteins, p51 induced apoptosis most significantly than p53 or p73 in melanoma cells. SK-mel-118 melanoma cells infected with p51-expressing adenovirus(Ad-p51) showed fragmented cellular DNA and activated caspase-3 characteristic of apoptosis. We prepared RNA from Ad-LacZ(control)-and Ad-p51-infected SK-mel-118 cells and compared amount of RNA by the methods of differential-display(DD)-PCR and DNA microarray. As a result, we identified more than twenty cellular genes that were clearly induced by p51. Inducibility of the candidate genes were examined in primary fibroblasts, keratinocytes and melanocytes by RT-PCR. Among them, three genes were induced in keratinocytes but not in fibroblasts or melanocytes. From these results, it is suggested that multiple genes including our candidates are induced by p51 and may have some roles in the process of cutaneous formation.