Investigation of ex vivo Shoc2 Ablation in Murine Keratinocytes

Sarı S., Rodriguez-Viciana P.

4th International Eurasian Conference on Science, Engineering and Technology, Ankara, Turkey, 14 - 16 December 2022, pp.206

  • Publication Type: Conference Paper / Summary Text
  • City: Ankara
  • Country: Turkey
  • Page Numbers: pp.206
  • Abdullah Gül University Affiliated: Yes


Investigation of ex vivo Shoc2 Ablation in Murine Keratinocytes Sibel Sari* (ORCID: and Pablo Rodriguez-Viciana (ORCID: 0002-8846-543X) *1Abdullah Gul University, Faculty of Life and Natural Sciences, Molecular Biology and Genetics, Kayseri, Turkey. 2University College London (UCL) Cancer Institute, London, UK *Corresponding author e-mail: Abstract The skin has a complex network of different cell types that act dynamic cellular communication to maintain several important processes such as inflammation, wound healing, and angiogenesis. The multi-functionality of skin is supported by its organised structure. Its structure is composed of three layers, the epidermis, dermis and hypodermis. The epidermis is mainly populated by keratinocytes, along with melanocytes, immune cells and Langerhans cells. The RAS/RAF/MEK/ERK signalling is known to be required for proliferation of murine keratinocytes in vitro. Dephosphorylation of a conserved inhibitory site on RAF by a phosphatase complex comprised of SHOC2, MRAS and PP1 is one of the key steps in the process of RAF activation. Our previous results show that systemic inactivation of SHOC2 in adult mice results in skin dermatitis as well as a range of histopathological alterations that include epidermal hyperproliferation with hyperkeratosis, an increased number of anagen phase hair follicles. The SHOC2 KO skin also developed an extensive dermal and epidermal inflammatory infiltrate containing a variety of immune cells. Here, we sought to examine the effect of Shoc2 deletion in epidermal keratinocytes in cultured primary murine keratinocytes in vitro. To explore the functions of Shoc2 in keratinocytes, we prepared primary keratinocyte cultures from the tails of adult Shoc2fl/fl Rosa26-CreERT2 mice. We examined the role of acute Shoc2 inactivation on the proliferation of keratinocytes by performing Incucyte growth curves. As a result, although in vitro proliferation assays show that keratinocytes display defective proliferation in the absence of Shoc2, Shoc2 ablation in vivo leads to epidermal hyperplasia, with increased proliferation and defective differentiation of keratinocytes in the skin. These results strongly suggest that the epidermal hyperproliferation observed in vivo is not a keratinocyte autonomous effect but rather is likely a consequence of inflammation driven by immune cells. Future studies are needed to address this possibility. Keywords: ex vivo models, Shoc2, keratinocytes