Ji Young Oh1,2, Han Na Suh2,3, Gee Euhn Choi2, Hyun Jik Lee2, Young Hyun Jung2, So Hee Ko2, Jun Sung Kim2, Chang Woo Chae2, Chang-Kyu Lee1,4, Ho Jae Han2. 1. Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute of Agriculture and Life Science, Seoul National University, Seoul, Korea. 2. Department of Veterinary Physiology, College of Veterinary Medicine, Research Institute for Veterinary Science, and BK21 PLUS Program for Creative Veterinary Science Research, Seoul National University, Seoul, Korea. 3. Minipig Model Group, Animal Model Center, Korea Institute of Toxicology, Jeongeup-si, Jeollabuk-do, Korea. 4. Institute of Green Bio Science and Technology, Seoul National University, Pyeong Chang, Kangwon do, Korea.
Abstract
BACKGROUND AND PURPOSE: The sonic hedgehog pathway (Shh) plays a central role in maintaining stem cell function and behaviour in various processes related to self-renewal and tissue regeneration. However, the therapeutic effect of Shh on mouse embryonic stem cells (mESCs) has not yet been clearly elucidated. Thus, we investigated the effect of Shh on the regulation of mESC behaviour as well as the effect of Shh-pretreated mESCs in skin wound healing. EXPERIMENTAL APPROACH: The underlying mechanisms of Shh signalling pathway in growth and motility of mESCs were investigated using Western blot analysis, a cell proliferation assay and cell migration assay. In addition, the effect of Shh-pretreated mESCs in skin wound healing was determined using a mouse excisional wound splinting model. KEY RESULTS: Shh disrupted the adherens junction through proteolysis by activating MMPs. In addition, the release of β-catenin from adherens junctions mediated by Shh led to cell cycle-dependent mESC proliferation. Shh-mediated Gli1 expression led to integrin β1 up-regulation, followed by FAK and Src phosphorylation. Furthermore, among the Rho-GTPases, Rac1 and Cdc42 were activated in a Shh-dependent manner while F-actin expression was suppressed by Rac1 and Cdc42 siRNA transfection. Consistent with the in vitro results, the skin wound healing assay revealed that Shh-treated mESCs increased angiogenesis and skin wound repair compared to that in Shh-treated mESCs transfected with integrin β1 siRNA in vivo. CONCLUSIONS AND IMPLICATIONS: Our results imply that Shh induces adherens junction disruption and integrin β1-dependent F-actin formation by a mechanism involving FAK/Src and Rac1/Cdc42 signalling pathways in mESCs.
BACKGROUND AND PURPOSE: The sonic hedgehog pathway (Shh) plays a central role in maintaining stem cell function and behaviour in various processes related to self-renewal and tissue regeneration. However, the therapeutic effect of Shh on mouse embryonic stem cells (mESCs) has not yet been clearly elucidated. Thus, we investigated the effect of Shh on the regulation of mESC behaviour as well as the effect of Shh-pretreated mESCs in skin wound healing. EXPERIMENTAL APPROACH: The underlying mechanisms of Shh signalling pathway in growth and motility of mESCs were investigated using Western blot analysis, a cell proliferation assay and cell migration assay. In addition, the effect of Shh-pretreated mESCs in skin wound healing was determined using a mouse excisional wound splinting model. KEY RESULTS:Shh disrupted the adherens junction through proteolysis by activating MMPs. In addition, the release of β-catenin from adherens junctions mediated by Shh led to cell cycle-dependent mESC proliferation. Shh-mediated Gli1 expression led to integrin β1 up-regulation, followed by FAK and Src phosphorylation. Furthermore, among the Rho-GTPases, Rac1 and Cdc42 were activated in a Shh-dependent manner while F-actin expression was suppressed by Rac1 and Cdc42 siRNA transfection. Consistent with the in vitro results, the skin wound healing assay revealed that Shh-treated mESCs increased angiogenesis and skin wound repair compared to that in Shh-treated mESCs transfected with integrin β1 siRNA in vivo. CONCLUSIONS AND IMPLICATIONS: Our results imply that Shh induces adherens junction disruption and integrin β1-dependent F-actin formation by a mechanism involving FAK/Src and Rac1/Cdc42 signalling pathways in mESCs.
Authors: Huong Le; Rebecca Kleinerman; Oren Z Lerman; Daniel Brown; Robert Galiano; Geoffrey C Gurtner; Stephen M Warren; Jamie P Levine; Pierre B Saadeh Journal: Wound Repair Regen Date: 2008 Nov-Dec Impact factor: 3.617
Authors: Young A Yoo; Myoung Hee Kang; Hyun Joo Lee; Baek-hui Kim; Jong Kuk Park; Hyun Koo Kim; Jun Suk Kim; Sang Cheul Oh Journal: Cancer Res Date: 2011-10-05 Impact factor: 12.701
Authors: Mi Ok Kim; Jung Min Ryu; Han Na Suh; Soo Hyun Park; Yeon-Mok Oh; Sang Hun Lee; Ho Jae Han Journal: Stem Cells Dev Date: 2015-09-02 Impact factor: 3.272
Authors: Ji Young Oh; Han Na Suh; Gee Euhn Choi; Hyun Jik Lee; Young Hyun Jung; So Hee Ko; Jun Sung Kim; Chang Woo Chae; Chang-Kyu Lee; Ho Jae Han Journal: Br J Pharmacol Date: 2018-07-26 Impact factor: 8.739