Up-Regulation of 14-3-3σ in Hacat upon Co-Cultivation with HEPM
- *Corresponding Author:
- Cuong Hoang-Vu
Department of General
Visceral and Vascular Surgery
Martin-Luther-University Halle-Wittenberg, Germany
E-mail: [email protected]
Received Date: January 07, 2012; Accepted Date: April 12, 2013; Published Date: April 18, 2013
Citation: Magnucki G, Bialek J, Hammje K, Prenzel H, Klohs G, et al. (2013) Up-Regulation of 14-3-3s in Hacat upon Co-Cultivation with HEPM. J Clin Exp Dermatol Res 4:171. doi: 10.4172/2155-9554.1000171
Copyright: © 2013 Magnucki G, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Background: To generate an in vitro system of the dermal epithelial-mesenchymal interaction we coÃ¢ÂÂcultured HaCaT (keratinocytes) and HEPM (embryonic mesenchyme). This model allowed a local disjunction with preserved cell-cell communication.
Methods: For the analysis of different protein expression patterns between co- and pure-cultured HaCaTs we performed 2D-electrophoresis and mass spectrometry. Afterwards the mass spectromertically identified protein 14-3- 3σ was silenced by siRNA in HaCaT cells.
Results: We analyzed 28 spots and found 17 different expressed mainly metabolic and cytoskeletal proteins. Interestingly, stratifin (14-3-3σ), maspin and Profilin-1 (PFN1) were up-regulated, whereas Peroxiredoxin-5 (PRDX5), PDZ-and-LIM-domain-protein-1 (CLIM1) and Annexin A1 (ANXA1) were decreased in co-cultured HaCaTs. The specific knock-down of 14-3-3σ resulted in a down-regulation of RhoA, Rac1/2/3, LIMK1 and phosphorylated cofilin, which are involved in cytoskeleton dynamics. Furthermore, reduction of 14-3-3σ coincided with significantly decreased proliferation rates and levels of Ki67.
Conclusions: We concluded that the interaction with mesenchymal cells may initiate the alternation of epidermal precursor cells to differentiated keratinocytes, what was confirmed by the up-regulation of different keratins. Furthermore, 14-3-3σ may influence the proliferation-rate of keratinocytes via Rho GTPases.