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Knockdown of PKD1 in normal human epidermal keratinocytes increases mRNA expression of keratin 10 and involucrin: early markers of keratinocyte differentiation

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Abstract

Subconfluent normal human keratinocytes exhibit autonomous (autocrine growth factor driven) proliferation and express the specific markers for keratinocyte proliferation K5 (keratin 5) and K14 (keratin 14). Utilizing this model the effects of PKD1 (Protein kinase D1) knockdown on activation of differentiation was studied. siRNA approach was applied to achieve specific knockdown of PKD1 and the mRNA levels of different keratinocyte markers—K14 and PCNA (markers of basal proliferating keratinocytes), involucrin and K10 (early differentiation markers) were analyzed. Treatment of cultured keratinocytes with siRNA for PKD1 resulted in reduction of mRNA levels of PKD1, altered cell phenotype and promotion of keratinocyte differentiation, demonstrated by increased expression of involucrin and K10 mRNAs. No significant changes in K14 mRNA expression levels were detected, but the expression of PCNA mRNA was markedly diminished. This study was the first to show that mRNA expression of PKD1 in subconfluent normal human keratinocytes is very low, the PKD1 mRNA levels were more than 8-fold lower than the same ones in hTert keratinocytes. These findings suggest antidifferentiative role of PKD1 in normal human keratinocytes, contrary to the prodiferentiative role of PKD1 in human hTert keratinocytes. We came to the conclusion that there are differences between transduction pathways involving PKD1 in primary human keratinocyte cultures and these in immortalized hTert keratinocytes.

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Acknowledgements

This work was supported by the Ministry of Education and Science—Bulgaria Grant No. BY 03-2005 and by a Grant from Medical University of Sofia. We thank F. Minner (LabCeTi, FUNDP, Namur-Belgium) for the construction of the primers for PKD1 and PCNA and F. Herphelin (LabCeTi, FUNDP, Namur-Belgium) for her technical help and useful discussions.

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Authors and Affiliations

  1. Department “Chemistry and Biochemistry”, Medical University, 2 Zdrave str., 1431, Sofia, Bulgaria

    Petya Ivanova, Ganka Atanasova & Vanyo Mitev

  2. Cell and Tissues Laboratory, URPHYM, University of Namur (FUNDP), 61 Rue De Bruxelles, 5000, Namur, Belgium

    Ganka Atanasova & Yves Poumay

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  1. Petya Ivanova
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  2. Ganka Atanasova
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  4. Vanyo Mitev
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Correspondence to Vanyo Mitev.

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Ivanova, P., Atanasova, G., Poumay, Y. et al. Knockdown of PKD1 in normal human epidermal keratinocytes increases mRNA expression of keratin 10 and involucrin: early markers of keratinocyte differentiation . Arch Dermatol Res 300, 139–145 (2008). https://doi.org/10.1007/s00403-008-0832-7

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  • DOI: https://doi.org/10.1007/s00403-008-0832-7

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