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KMID : 0191120140290020265
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Journal of Korean Medical Science
2014 Volume.29 No. 2 p.265 ~ p.271
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Microarray Analysis for Genes Associated with Angiogenesis in Diabetic OLETF Keratocytes
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Park Jun-Mo
Park Young-Min
Jung Wook
Lee Ji-Eun
Lee Jong-Soo
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Abstract
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The purpose of this study was to identify the differences in angiogenesis gene expression between normal and diabetic keratocytes stimulated with interleukin-1α (IL-1α) and tumor necrosis factor-α (TNF-α). Primarily cultured normal and diabetic keratocytes were treated with 20 ng/mL of IL-1a and TNF-α for 6 hr. cDNA was hybridized to an oligonucleotide microarray. Microarray analysis was used to identify differentially expressed genes that were further evaluated by real-time polymerase chain reaction (RT-PCR). Diabetes keratocytes overexpressed vital components of angiogenesis including Agtr1, and under-expressed components related to the blood vessel maturation, including Dcn. Cytokine-treated diabetic keratocytes differentially expressed components of angiogenesis. OLETF keratocytes after treatment with IL-1α and TNF-α showed the newly expressed 15 and 14 genes, respectively. Newly and commonly under-expressed five genes followed by treatment with both IL-1α and TNF-α were also evident. RT-PCR showed results similar to the microarray results. Agtr1 and Itga1 showed an increased expression in diabetic keratocytes compared with normal corneal keratocytes, especially after TNF-α treatment. Il6 appeared strong expression after interleukin-1α treatment, but showed down expression after TNF-α treatment. Further studies to analyze and confirm the significance of the identified angiogenetic genes of diabetes are needed.
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KEYWORD
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Angiogenesis, Cytokines, Inflammation, Corneal Keratocyte, Microarray Analysis, OLETF
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