title: Chondrocyte-derived extracellular matrix suppresses pathogenesis of human pterygium epithelial cells by blocking the NF-κB signaling pathways.
abstract: We previously have reported that chondrocyte-derived extracellular matrix (CDECM) suppresses the growth of pterygium in athymic nude mice. The aim of this study is to demonstrate the effect of CDECM on the pterygium epithelial cells and molecular signaling pathways in human primary pterygium epithelial cells (hPECs).
  
  
  
  Human conjunctival epithelial cells (hConECs) were used for identification of the effect of CDECM on normal conjunctiva. The effects of CDECM on proliferation were measured with the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfenyl)-2H-tetrazolium (MTS) assay. Cell migration was evaluated according to the scratch wound closure assay and the Transwell invasion assay. Pterygium-related angiogenesis, inflammation, and extracellular matrix remodeling were analyzed with immunoblot and enzyme-linked immunosorbent assay (ELISA). The level of oxidative stress was detected with 2',7'-dichlorofluorescein diacetate (DCFH-DA). Protein kinase signaling was also analyzed with immunoblot.
  
  
  
  CDECM did not show cytotoxicity until 1 mg/ml in the hConECs and hPECs. Cell migration and invasion were markedly reduced by treatment of 1 mg/ml CDECM in the hPECs to 34% of the control, but not in the hConECs. CDECM significantly downregulated matrix metallopeptidase 9 (MMP-9) and fibronectin and upregulated tissue inhibitor of metalloprotease 1 (TIMP-1) and -2 in the hPECs. Angiogenic factors, such as vascular endothelial growth factor (VEGF), antivascular cellular adhesion molecule 1 (VCAM-1), and cluster of differentiation 31 (CD31), and proinflammatory factors, including tumor necrosis factor-α (TNF-α), cyclooxygenase-2 (Cox2), interleukin 6 (IL-6), and prostaglandin E2(PGE2), were dramatically reduced by CDECM in the hPECs. Furthermore, CDECM significantly inhibited the generation of intracellular reactive oxygen species and the expression of NADPH oxidase subunits, Nox2 and p47phox. CDECM induced nuclear factor erythroid-2 related factor 2 (Nrf2) mediated-antioxidant enzyme heme oxygenase-1 (HO-1). CDECM also suppressed nuclear factor-kappa B (NF-κB) activation and the phosphorylation of p38 mitogen-activated protein kinase (MAPK), protein kinase C alpha (PKCα), and PKCθ.
  
  
  
  CDECM was markedly effective in pathogenesis of hPECs. CDECM-suppressed migration of hPECs resulted from the inhibition of NF-κB activation and the improvement of Nrf2 induction by blocking the p38 MAPK and PKC signaling pathways.
authors.affiliation: Ocular Neovascular Disease Research Center, Inje University Busan Paik Hospital, Busan, Republic of Korea.
authors.forename: Hyesook
authors.initials: H
authors.lastname: Lee
authors.affiliation: Ocular Neovascular Disease Research Center, Inje University Busan Paik Hospital, Busan, Republic of Korea.
authors.forename: Minsup
authors.initials: M
authors.lastname: Lee
authors.affiliation: Ocular Neovascular Disease Research Center, Inje University Busan Paik Hospital, Busan, Republic of Korea; Department of Ophthalmology, Inje University College of Medicine, Busan, Republic of Korea.
authors.forename: Yoonjin
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Division of Industrial Technology, Korea Evaluation Institute of Industrial Technology, Daegu, Republic of Korea.
authors.forename: Soojin
authors.initials: S
authors.lastname: Choi
authors.affiliation: Ocular Neovascular Disease Research Center, Inje University Busan Paik Hospital, Busan, Republic of Korea; Department of Ophthalmology, Inje University College of Medicine, Busan, Republic of Korea.
authors.forename: Jaewook
authors.initials: J
authors.lastname: Yang
mesh_terms: D000368:Aged; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D019902:Chondrocytes / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology*; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D006801:Humans; D007249:Inflammation / Q000473:pathology; D008297:Male; D008875:Middle Aged; D008954:Models, Biological; D016328:NF-kappa B / Q000378:metabolism*; D018919:Neovascularization, Physiologic; D018384:Oxidative Stress; D011625:Pterygium / Q000473:pathology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D016328:NF-kappa B; D017382:Reactive Oxygen Species