Shotgun label‐free proteomic analysis for identification of proteins in HaCaT human skin keratinocytes regulated by the administration of collagen from soft‐shelled turtle

Soft‐shelled turtles (Pelodiscus sinensis) are widely distributed in some Asian countries, and we previously reported that soft‐shelled turtle tissue could be a useful material for collagen. In the present study, we performed shotgun liquid chromatography (LC)/mass spectrometry (MS)‐based global proteomic analysis of collagen‐administered human keratinocytes to examine the functional effects of collagen from soft‐shelled turtle on human skin. Using a semiquantitative method based on spectral counting, we were able to successfully identify 187 proteins with expression levels that were changed more than twofold by the administration of collagen from soft‐shelled turtle. Based on Gene Ontology analysis, the functions of these proteins closely correlated with cell–cell adhesion. In addition, epithelial–mesenchymal transition was induced by the administration of collagen from soft‐shelled turtle through the down‐regulation of E‐cadherin expression. Moreover, collagen‐administered keratinocytes significantly facilitated wound healing compared with nontreated cells in an in vitro scratch wound healing assay. These findings suggest that collagen from soft‐shelled turtle provides significant benefits for skin wound healing and may be a useful material for pharmaceuticals and medical care products. © 2017 The Authors Journal of Biomedical Materials Research Part B: Applied Biomaterials Published by Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2403–2413, 2018.