The Thai Journal of Veterinary Medicine


Involucrin (INV) is a well-known early stage marker of terminal keratinocyte differentiation in humans. Besides its importance in human cancer, it has been predicted to be a prognostic marker in canine skin cancer. However, there is very little information available on canine INV. Therefore, for the first time, a three-dimensional (3D) cultured canine keratinocyte cell line was evaluated for the expression level of INV and two other cancer prognostic parameters, cytokeratin 10 (CK10) and Ki67, compared to that in canine normal skin (CN) and cutaneous squamous cell carcinoma (CSCC) cells. The INV protein and transcript expression levels were significantly (p < 0.005) higher in canine CN compared to in CSCC, whereas Ki67 expression levels were significantly (p < 0.05) lower in CN than in CSCC. The culture of a commercial canine keratinocyte cell line (CPEK; CELLnTEC Advanced Cell Systems, Switzerland) to produce the 3D epidermal skin was successfully achieved under an air-liquid interface for 14 d of culture. There was no statistical difference in the INV and CK10 expression levels between the 3D-cultured CPEK cells and CN. A custom designed INV siRNA transfected at 300 pmol showed the high efficacy of INV knockdown with minimum cytotoxicity to cultured CPEK cells, significantly reducing the INV mRNA expression level after 5 h transfection for up to 48 h. This is the first report on the custom design of siRNA transfection in monolayer cultured CPEK cells for INV gene knockdown in dogs and it could be useful for future studies on the carcinogenesis of CSCC.

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