Molecular characterization and expression of p63 isoforms in human keloids

Keloids are benign skin tumors that develop following wounding. A cDNA product from human keloid specimens was identified using the differential display technique. The full-length cDNA was cloned by RT-PCR using human keloid mRNA as template. The predicted product of the cDNA was found to be 99% identical to the DeltaN-p63 gamma isotype of p63, a transcription factor that belongs to the family that includes the structurally related tumor suppressor p53 and p73. The DeltaN-p63 isotype lacks the acidic N terminal region corresponding to the transactivation domain of p53. Since this can potentially block p53-mediated target gene transactivation, it may serve as a dominant-negative isoform. Real-Time RT-PCR analysis of RNAs from normal skin tissue and keloids showed that the DeltaN-p63 isotype is specifically expressed in keloids, but is virtually undetectable in normal skin. Immunostaining of p63 in normal skin revealed that only basal cells of the epithelium expressed the protein, while in keloid tissues the antigen was detected in the nuclei of cells scattered through all layers of the epithelium and in fibroblast-like cells in the dermis. These results may indicate that aberrant p63 expression plays a role not only in malignant tumors but also in benign skin diseases that show hyperproliferation of epidermal cells in vivo. Moreover, this isoform of p63 could serve as a specific molecular marker for this human disease.