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Induced expression of insulin-like growth factor-1 by amniotic membrane-conditioned medium in cultured human corneal epithelial cells

 Joon H. Lee  ;  Ik Hee Ryu  ;  Eung Kweon Kim  ;  Jong Eun Lee  ;  SoonWon Hong  ;  Hyung Keun Lee 
 Investigative Ophthalmology & Visual Science, Vol.47(3) : 864-872, 2006 
Journal Title
 Investigative Ophthalmology & Visual Science 
Issue Date
Amnion/physiology* ; Blotting, Western ; Cell Proliferation/drug effects ; Cells, Cultured ; Culture Media, Conditioned/pharmacology* ; DNA/biosynthesis ; Dose-Response Relationship, Drug ; Epithelium, Corneal/cytology* ; Epithelium, Corneal/metabolism* ; Female ; Humans ; Insulin-Like Growth Factor I/biosynthesis* ; Insulin-Like Growth Factor I/genetics ; Phosphorylation ; Proto-Oncogene Proteins c-akt/metabolism ; RNA, Messenger/metabolism ; RNA, Small Interfering/pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Tetrazolium Salts ; Thiazoles ; Time Factors ; Up-Regulation
To determine the effect of amniotic membrane-conditioned medium (AMCM), via insulin-like growth factor (IGF)-1 induction, on human corneal epithelial cell (HCEC) proliferation. METHODS: HCECs were cultured from corneal limbal tissue with supplemented hormonal epithelial medium (SHEM). After administration of AMCM, cell proliferation was evaluated with an MTT assay and DNA synthesis with methyl-[(3)H]-thymidine incorporation assay. RT-PCR and Western immunoblot analyses were performed, to determine potential inducible factors that may be associated with AMCM-induced cell proliferation. Neutralizing anti-IGF-1 antibody and small interfering (si)RNA were also used to clarify the role of IGF-1 in AMCM-induced HCEC proliferation. RESULTS: HCEC proliferation increased after AMCM treatment. Of the cytokines known to be associated with HCEC proliferation, only IGF-1 expression was upregulated in response to AMCM in a dose- and time-dependent manner. The IGF-1 induction effect was found on both AMCM from live AM and from cryopreserved AM. HCEC proliferation was also increased by addition of exogenous IGF-1. AMCM-induced HCEC proliferation was inhibited in the presence of neutralizing anti-IGF-1 antibody and IGF-1 siRNA. Finally, Akt phosphorylation was increased in HCECs after AMCM treatment and was inhibited by IGF-1 siRNA. CONCLUSIONS: IGF-1 is induced by AMCM during HCEC proliferation, and this induction may play an important role in the amniotic membrane during HCEC proliferation and migration in several intractable corneal epithelial defects.
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1. College of Medicine (의과대학) > Dept. of Ophthalmology (안과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Pathology (병리학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Anatomy (해부학교실) > 1. Journal Papers
Yonsei Authors
Kim, Eung Kweon(김응권) ORCID logo https://orcid.org/0000-0002-1453-8042
Lee, Jong Eun(이종은) ORCID logo https://orcid.org/0000-0001-6203-7413
Lee, Joon Haeng(이준행)
Lee, Hyung Keun(이형근) ORCID logo https://orcid.org/0000-0002-1123-2136
Hong, Soon Won(홍순원) ORCID logo https://orcid.org/0000-0002-0324-2414
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