title: Galectin-1 stimulates motility of human umbilical cord blood-derived mesenchymal stem cells by downregulation of smad2/3-dependent collagen 3/5 and upregulation of NF-κB-dependent fibronectin/laminin 5 expression.
abstract: Galectin-1 (Gal-1) belongs to a family of endogenous lectins with conserved carbohydrate recognition domains binding β-galactosidase sugars and plays a vital role in regulating stem cell functions including determination of cell fate. However, our understanding of the functional roles of Gal-1 in human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) is still fragmentary and incomplete. Gal-1 significantly increased motility after a 24-h incubation, and this effect was inhibited by β-lactose. We analyzed 17 extracellular matrix (ECM) genes in UCB-MSCs. Gal-1 decreased the expression of collagen genes COL3A1 (COL-3) and COL5A1 (COL-5) but increased the expression of fibronectin (FN) and laminin 5 (LM-5), that were reversed by β-lactose. Gal-1 increased protein kinase C (PKC), c-Src, and caveolin-1 (Cav-1) phosphorylation that was attenuated by β-lactose and the Src inhibitor PP2. In addition, pretreatment with the lipid raft disruptor Mβ-CD and the PKC inhibitors inhibited Gal-1-induced UCB-MSC motility. In addition, Gal-1 reduced smad2/3 phosphorylation and induced nuclear factor (NF)-κB phosphorylation. Pretreatment with Mβ-CD attenuated Gal-1-reduced smad2/3 phosphorylation, COL-3, and COL-5 expression but did not affect NF-κB phosphorylation, FN, or LM-5 expression. In contrast, PKC inhibitors only attenuated NF-κB phosphorylation, FN, and LM-5 expression. Reconstructing Gal-1-induced genetic changes by replacing it with siRNA specific for COL-3 or COL-5, or treatment of the cells with FN and LM-5 proteins, increased motility and its related proteins such as focal adhesion kinase, Akt, Erk, integrins, and matrix metalloproteinase-2. A combined treatment with COL-3/COL-5 siRNA or FN/LM-5 compared with that of single treatments was synergistic. However, a single Gal-1 treatment maximally stimulated motility and related protein phosphorylation/expression. These results demonstrate that Gal-1 stimulated human UCB-MSC motility by decreasing COL-3/COL-5 expression and increasing FN/LM-5 expression through a PKC-dependent NF-κB and c-Src/Cav-1-dependent smad2/3 pathway that was critical for governing the activation of FAK, Akt, Erk, integrins, and MMP2.
authors: 1] Department of Veterinary Physiology, College of Veterinary Medicine, Research Institute for Veterinary Science, Seoul National University, Seoul, Korea [2] BK21 PLUS Creative Veterinary Research Center, Seoul National University, Seoul, Korea.
authors: S P
authors: SP
authors: Yun
authors: 1] Department of Veterinary Physiology, College of Veterinary Medicine, Research Institute for Veterinary Science, Seoul National University, Seoul, Korea [2] BK21 PLUS Creative Veterinary Research Center, Seoul National University, Seoul, Korea.
authors: S-J
authors: SJ
authors: Lee
authors: 1] Department of Veterinary Physiology, College of Veterinary Medicine, Research Institute for Veterinary Science, Seoul National University, Seoul, Korea [2] BK21 PLUS Creative Veterinary Research Center, Seoul National University, Seoul, Korea.
authors: Y H
authors: YH
authors: Jung
authors: 1] Department of Veterinary Physiology, College of Veterinary Medicine, Research Institute for Veterinary Science, Seoul National University, Seoul, Korea [2] BK21 PLUS Creative Veterinary Research Center, Seoul National University, Seoul, Korea.
authors: H J
authors: HJ
authors: Han
mesh_terms: D015815:Cell Adhesion Molecules / Q000235:genetics* / Q000378:metabolism; D002465:Cell Movement; D002478:Cells, Cultured; D024061:Collagen Type III / Q000235:genetics* / Q000378:metabolism; D024062:Collagen Type V / Q000235:genetics* / Q000378:metabolism; D015536:Down-Regulation; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism; D005353:Fibronectins / Q000235:genetics* / Q000378:metabolism; D037483:Galectin 1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D051899:Smad2 Protein / Q000235:genetics / Q000378:metabolism*; D051900:Smad3 Protein / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation; D000097794:Kalinin
chemical_list: D015815:Cell Adhesion Molecules; D024061:Collagen Type III; D024062:Collagen Type V; D005353:Fibronectins; D037483:Galectin 1; D016328:NF-kappa B; D051899:Smad2 Protein; D051900:Smad3 Protein; D000097794:Kalinin; C439971:COL3A1 protein, human; C530725:COL5A1 protein, human; C497782:SMAD2 protein, human; C497790:SMAD3 protein, human