Involucrin, cytokeratin 10 and Ki67 expression in a threedimensional cultured canine keratinocyte cell line in comparison to canine skin and cutaneous squamous cell carcinoma and a pilot study on custom-designed siRNA-INV transfection

  • Nongnut Assawawongkasem
  • Somporn Techangamsuwan
  • Prapruddee Piyaviriyakul
  • Pranom Puchadapirom
  • Achariya Sailasuta
Keywords: canine keratinocyte cell line, cutaneous squamous cell carcinoma, INV, CK10, Ki67

Abstract

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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Published
2020-04-24
How to Cite
Assawawongkasem, N., Techangamsuwan, S., Piyaviriyakul, P., Puchadapirom, P., & Sailasuta, A. (2020). Involucrin, cytokeratin 10 and Ki67 expression in a threedimensional cultured canine keratinocyte cell line in comparison to canine skin and cutaneous squamous cell carcinoma and a pilot study on custom-designed siRNA-INV transfection. he hai ournal of eterinary edicine, 50(1), 43-55. etrieved from https://he01.tci-thaijo.org/index.php/tjvm/article/view/243256
Section
Original Articles