title: Effect of Wnt3a on keratinocytes utilizing in vitro and bioinformatics analysis.
abstract: Wingless-type (Wnt) signaling proteins participate in various cell developmental processes. A suppressive role of Wnt5a on keratinocyte growth has already been observed. However, the role of other Wnt proteins in proliferation and differentiation of keratinocytes remains unknown. Here, we investigated the effects of the Wnt ligand, Wnt3a, on proliferation and differentiation of keratinocytes. Keratinocytes from normal human skin were cultured and treated with recombinant Wnt3a alone or in combination with the inflammatory cytokine, tumor necrosis factor α (TNFα). Furthermore, using bioinformatics, we analyzed the biochemical parameters, molecular evolution, and protein-protein interaction network for the Wnt family. Application of recombinant Wnt3a showed an anti-proliferative effect on keratinocytes in a dose-dependent manner. After treatment with TNFα, Wnt3a still demonstrated an anti-proliferative effect on human keratinocytes. Exogenous treatment of Wnt3a was unable to alter mRNA expression of differentiation markers of keratinocytes, whereas an altered expression was observed in TNFα-stimulated keratinocytes. In silico phylogenetic, biochemical, and protein-protein interaction analysis showed several close relationships among the family members of the Wnt family. Moreover, a close phylogenetic and biochemical similarity was observed between Wnt3a and Wnt5a. Finally, we proposed a hypothetical mechanism to illustrate how the Wnt3a protein may inhibit the process of proliferation in keratinocytes, which would be useful for future researchers.
authors.affiliation: Institute for Skeletal Aging & Orthopedic Surgery, Hallym University-Chuncheon Sacred Heart Hospital, Chuncheon 200704, Korea. jsnam88@hallym.ac.kr.
authors.forename: Ju-Suk
authors.initials: JS
authors.lastname: Nam
authors.affiliation: Institute for Skeletal Aging & Orthopedic Surgery, Hallym University-Chuncheon Sacred Heart Hospital, Chuncheon 200704, Korea. drchiranjib@yahoo.com.
authors.forename: Chiranjib
authors.initials: C
authors.lastname: Chakraborty
authors.affiliation: Institute for Skeletal Aging & Orthopedic Surgery, Hallym University-Chuncheon Sacred Heart Hospital, Chuncheon 200704, Korea. boneresearch@hallym.ac.kr.
authors.forename: Ashish Ranjan
authors.initials: AR
authors.lastname: Sharma
authors.affiliation: Department of Dermatology, School of Medicine, Kangwon National University Hospital, Chuncheon 200722, Korea. young6078@hanmail.net.
authors.forename: Young
authors.initials: Y
authors.lastname: Her
authors.affiliation: Institute for Skeletal Aging & Orthopedic Surgery, Hallym University-Chuncheon Sacred Heart Hospital, Chuncheon 200704, Korea. grant903@gmail.com.
authors.forename: Kee-Jeong
authors.initials: KJ
authors.lastname: Bae
authors.affiliation: Institute for Skeletal Aging & Orthopedic Surgery, Hallym University-Chuncheon Sacred Heart Hospital, Chuncheon 200704, Korea. microbio.garima@gmail.com.
authors.forename: Garima
authors.initials: G
authors.lastname: Sharma
authors.affiliation: Medical Biotechnology Division, School of Biosciences and Technology, VIT University, Vellore 632014, Tamil Nadu, India. georgecp77@yahoo.co.in.
authors.forename: George Priya
authors.initials: GP
authors.lastname: Doss
authors.affiliation: Institute for Skeletal Aging & Orthopedic Surgery, Hallym University-Chuncheon Sacred Heart Hospital, Chuncheon 200704, Korea. 123sslee@gmail.com.
authors.forename: Sang-Soo
authors.initials: SS
authors.lastname: Lee
authors.affiliation: Institute for Skeletal Aging & Orthopedic Surgery, Hallym University-Chuncheon Sacred Heart Hospital, Chuncheon 200704, Korea. bone2011@naver.com.
authors.forename: Myung-Sun
authors.initials: MS
authors.lastname: Hong
authors.affiliation: Institute for Skeletal Aging & Orthopedic Surgery, Hallym University-Chuncheon Sacred Heart Hospital, Chuncheon 200704, Korea. DKsong@hallym.ac.kr.
authors.forename: Dong-Keun
authors.initials: DK
authors.lastname: Song
mesh_terms: D000595:Amino Acid Sequence; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival; D002478:Cells, Cultured; D019295:Computational Biology; D019143:Evolution, Molecular; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D015603:Keratinocytes / Q000166:cytology*; D019394:Ki-67 Antigen / Q000096:biosynthesis / Q000235:genetics; D010802:Phylogeny; D060066:Protein Interaction Maps; D011518:Proto-Oncogene Proteins / Q000235:genetics; D011565:Psoriasis / Q000473:pathology; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000494:pharmacology; D016415:Sequence Alignment; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D051153:Wnt Proteins / Q000235:genetics; D060449:Wnt Signaling Pathway / Q000235:genetics; D000071818:Wnt-5a Protein; D060509:Wnt3A Protein / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*
chemical_list: D019394:Ki-67 Antigen; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; C555510:WNT3A protein, human; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein; D060509:Wnt3A Protein