title: Negative pressure induces p120-catenin-dependent adherens junction disassembly in keratinocytes during wound healing.
abstract: A negative-pressure of 125mmHg (NP) has been widely used to treat chronic wounds in modern medicine. Keratinocytes under NP treatment have shown accelerated cell movement and decreased E-cadherin expression. However, the molecular mechanism of E-cadherin regulation under NP remains incompletely understood. Therefore, we investigated the E-cadherin regulation in keratinocytes (HaCaT cells) under NP. HaCaT cells were treated at ambient pressure (AP) and NP for 12h. Cell movement was measured by traditional and electric wound healing assays at the 2 different pressures. Mutants with overexpression of p120-catenin (p120(ctn)) were used to observe the effect of NP on p120(ctn) and E-cadherin expression during wound healing. Cell fractionation and immunoblotting data showed that NP increased Y228-phosphorylated p120(ctn) level and resulted in the translocation of p120(ctn) from the plasma membrane to cytoplasm. Immunofluorescence images revealed that NP decreased the co-localization of p120(ctn) and E-cadherin on the plasma membrane. Knockdown of p120(ctn) reduced E-cadherin expression and accelerated cell movement under AP. Overexpression of the Y228-phosphorylation-mimic p120(ctn) decreased E-cadherin membrane expression under both AP and NP. Phosphorylation-deficient mutants conferred restored adherens junctions (AJs) under NP. The Src inhibitor blocked the phosphorylation of p120(ctn) and impeded cell migration under NP. In conclusion, Src-dependent phosphorylation of p120(ctn) can respond rapidly to NP and contribute to E-cadherin downregulation. The NP-induced disassembly of the AJ further accelerates wound healing.
authors.affiliation: Healthy Aging Research Center, Graduate Institute of Rehabilitation Science, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Ching-Hui
authors.initials: CH
authors.lastname: Huang
authors.affiliation: Department of Physical Medicine and Rehabilitation, Chang Gung Memorial Hospital, Keelung, Taiwan; School of Traditional Chinese Medicine, College of Medicine, Chang Gung University, Taoyuan, Taiwan. Electronic address: steele0618@gmail.com.
authors.forename: Chih-Chin
authors.initials: CC
authors.lastname: Hsu
authors.affiliation: Department of Physical Medicine and Rehabilitation, Chang Gung Memorial Hospital, Taipei, Taiwan.
authors.forename: Carl Pai-Chu
authors.initials: CP
authors.lastname: Chen
authors.affiliation: Center of General Studies, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Shu-Er
authors.initials: SE
authors.lastname: Chow
authors.affiliation: Healthy Aging Research Center, Graduate Institute of Rehabilitation Science, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Jong-Shyan
authors.initials: JS
authors.lastname: Wang
authors.affiliation: Community Medicine Research Center, Chang Gung Memorial Hospital, Keelung, Taiwan; Institute of Molecular Biology, Academia Sinica, Nankang, Taipei, Taiwan.
authors.forename: Yu-Chiau
authors.initials: YC
authors.lastname: Shyu
authors.affiliation: Community Medicine Research Center, Chang Gung Memorial Hospital, Keelung, Taiwan.
authors.forename: Mu-Jie
authors.initials: MJ
authors.lastname: Lu
mesh_terms: D022005:Adherens Junctions / Q000378:metabolism*; D015820:Cadherins / Q000378:metabolism; D051177:Catenins / Q000378:metabolism*; D002460:Cell Line; D002465:Cell Movement; D015536:Down-Regulation; D055785:Gene Knockdown Techniques; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism* / Q000473:pathology*; D008954:Models, Biological; D010641:Phenotype; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011312:Pressure; D021381:Protein Transport; D013347:Subcellular Fractions / Q000378:metabolism; D014945:Wound Healing; D019061:src-Family Kinases / Q000378:metabolism; D000095002:Delta Catenin
chemical_list: D015820:Cadherins; D051177:Catenins; D019000:Phosphotyrosine; D019061:src-Family Kinases; D000095002:Delta Catenin; C000722347:CTNND1 protein, human
keywords: Cell movement; E-cadherin; Negative-pressure wound therapy; Wound healing