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Role of TGFBIp in Wound Healing and Mucin Expression in Corneal Epithelial Cells

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      Abstract

      Purpose

      Transforming growth factor-β-induced protein (TGFBIp) is highly expressed in the cornea, and mutant TGFBIp induces corneal diseases. However, the function of TGFBIp in cornea epithelium is not fully investigated. Here, we tested the importance of TGFBIp in regulation of gene expression and corneal epithelial cell (CEC) activity.

      Materials and Methods

      The effect of TGFBIp on CEC activity was analyzed by cell migration, adhesion, proliferation and wound healing assay. Analysis of gene expression was examined by western blot and quantitative reverse transcription PCR.

      Results

      The results demonstrated that TGFBIp increased adhesion, migration, proliferation, and wound healing of CECs. Analysis of gene expression presented that TGFBIp-stimulated CECs exhibited increased expression of mucin family genes, such as MUC1, -4, -5AC, and -16. Furthermore, TGFBIp treatment increased the expression of MUC1, -4, -5AC, -7, and -16 in conjunctival epithelial cells. TGFBIp also increased the activity of intracellular signaling molecules ERK and AKT in CECs. Using pharmacologic inhibitors of ERK and AKT, we showed that the expression of mucin genes by TGFBIp is mediated by the activation of ERK and AKT signaling.

      Conclusion

      Our findings demonstrate that the locally generated TGFBIp in the cornea may contribute to wound healing of CECs by enhancing the migration, adhesion, and proliferation of CECs. In addition, our results suggest that TGFBIp has a protective effect on ocular surfaces by inducing the expression of mucin genes in corneal and conjunctival epithelial cells. These data suggest that TGFBIp is a useful therapeutic target for patients with corneal wounds.

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      Most cited references 40

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          An SV40-immortalized human corneal epithelial cell line and its characterization.

          The authors attempted to immortalize human corneal epithelial cells; it is difficult to propagate primary human corneal epithelial cells because of scarcity of available tissue. However, cell immortalization by virus is always accompanied by shedding of free virus. The current study was performed to establish a cell line that produces no free viral particle. Primary cultured human corneal epithelial cells were infected with a recombinant sv40-adenovirus vector and were cloned three times to obtain a continuously growing cell line. Morphologic, cytologic, and biochemical characteristics of this cell line were analyzed. This cell line continued to grow for more than 400 generations, exhibiting a cobblestone-like appearance similar to normal corneal epithelial cells in culture. Transmission electron microscopy showed the evidence for the characteristic features of epithelial cells, including desmosome formation and development of microvilli. It expressed cornea-specific, 64-kD cytokeratin in addition to five major insoluble proteins. By enzymatic analysis using NADP as a coenzyme and a gas chromatograph mass spectrometer, this cell line was found to possess 8.71 IU/mg protein of aldehydedehydrogenase activity. When this cell line was grown at air-liquid interface on collagen type I gel, it differentiated in a multilayered fashion. The authors have established an SV40-immortalized human corneal epithelial cell line with properties similar to normal corneal epithelial cells.
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            Author and article information

            Affiliations
            [1 ]Department of Ophthalmology, Corneal Dystrophy Research Institute, Yonsei University College of Medicine, Seoul, Korea.
            [2 ]Institute of Vision Research, Severance Biomedical Science Institute, Yonsei University College of Medicine, Seoul, Korea.
            Author notes
            Corresponding author: Dr. Eung Kweon Kim, Department of Ophthalmology, Yonsei University College of Medicine, 50-1 Yonsei-ro, Seodaemun-gu, Seoul 03722, Korea. Tel: 82-2-2228-0824, Fax: 82-2-2227-8129, eungkkim@ 123456yuhs.ac
            Journal
            Yonsei Med J
            Yonsei Med. J
            YMJ
            Yonsei Medical Journal
            Yonsei University College of Medicine
            0513-5796
            1976-2437
            01 March 2017
            16 January 2017
            : 58
            : 2
            : 423-431
            28120575
            5290024
            10.3349/ymj.2017.58.2.423
            © Copyright: Yonsei University College of Medicine 2017

            This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

            Funding
            Funded by: National Research Foundation of Korea, CrossRef http://dx.doi.org/10.13039/501100003725;
            Award ID: NRF 2014R1A1A2057458
            Award ID: NRF-2016R1D1A1B03933337
            Funded by: Yonsei University College of Medicine, CrossRef http://dx.doi.org/10.13039/501100008005;
            Award ID: 6-2007-0173
            Categories
            Original Article
            Ophthalmology

            Medicine

            tgfbip, mucin, cornea, epithelial cells

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