Journal of the Korean Ophthalmological Society 2001;42(12):1760-1770.
Published online December 1, 2001.
The Differential Gene Expression in Corneal Fibroblast Cultured with Amniotic Membrane Components.
Young Jin Kim, Kyung Hwan Shyn, Jae Chan Kim
Department of Ophthalmology, College of Medicine, Chung-Ang University, Korea. jck50ey@kornet.net
양막성분으로 배양한 각막섬유모세포에서의 Differential Gene Expression
김영진(Young Jin Kim),신경환(Kyung Hwan Shyn),김재찬(Jae Chan Kim)
Abstract
PURPOSE
To know whether amniotic membrane (AM) extract could protect cell from toxic stimuli by changing gene expression such as inflammatory and apoptosis-related genes, cultured corneal fibroblasts were used. METHODS: After AM was pulverized in the liquid nitrogen and homogenized under the 4(C, it was centrifuged and its supernatant was obtained. Human corneal fibroblasts (HCFs) were divided into 2 groups - one group treated with AM extracts and TNF-alpha, and the other with TNF-alpha only. Cell morphology and viability were assessed and differential gene expression through cDNA array was performed. RESULTS: Viability of HCF cultured in AM extracts and TNF-alpha was better than that of cells in TNF-alpha only. Cells in TNF-alpha only were morphologically changed and dead. cDNA array showed the decrease in IL-1 , IL-1 receptor type 1 and caspases-1,-3,and -10 expression in cells treated with AM extracts. MMP-1,-2,-3 and-9 were almost the same or slightly increased and NF-kappaB, I kappaB kinase alpha, integrin-alpha1 and -alpha10, and MAP kinase were increased in cells with AM extracts. CONCLUSIONS: When cultured corneal fibroblasts were insulted by TNF-alpha, AM extracts seemed to protect the cells, by down-regulating various inflammation - and cell death- related genes, and by increasing the expression of genes enhancing cell proliferation and differentiation.
Key Words: Amniotic membrane;Corneal fibroblast;cDNA array;Differential gene expression


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