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) |
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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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