title: Microarray analysis for genes associated with angiogenesis in diabetic OLETF keratocytes.
abstract: 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.
authors.affiliation: Department of Ophthalmology, Busan St. Mary's Hospital, Busan, Korea.
authors.forename: Jun-Mo
authors.identifier: 0000-0002-3942-3989
authors.initials: JM
authors.lastname: Park
authors.affiliation: Department of Ophthalmology, School of Medicine, Pusan National University and Medical Research Institute, Pusan National University Hospital, Busan, Korea.
authors.forename: Young Min
authors.identifier: 0000-0001-8495-9224
authors.initials: YM
authors.lastname: Park
authors.affiliation: Department of Ophthalmology, School of Medicine, Pusan National University and Medical Research Institute, Pusan National University Hospital, Busan, Korea.
authors.forename: Wook
authors.identifier: 0000-0003-2364-9526
authors.initials: W
authors.lastname: Jung
authors.affiliation: Department of Ophthalmology, School of Medicine, Pusan National University and Medical Research Institute, Pusan National University Hospital, Busan, Korea.
authors.forename: Ji-Eun
authors.identifier: 0000-0002-4927-2854
authors.initials: JE
authors.lastname: Lee
authors.affiliation: Department of Ophthalmology, School of Medicine, Pusan National University and Medical Research Institute, Pusan National University Hospital, Busan, Korea.
authors.forename: Jong-Soo
authors.identifier: 0000-0002-1052-2924
authors.initials: JS
authors.lastname: Lee
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005786:Gene Expression Regulation / Q000187:drug effects; D053582:Interleukin-1alpha / Q000494:pharmacology; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D018919:Neovascularization, Physiologic / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D044140:Receptor, Angiotensin, Type 1 / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: C544067:Agtr1a protein, rat; D053582:Interleukin-1alpha; D044140:Receptor, Angiotensin, Type 1; D014409:Tumor Necrosis Factor-alpha
keywords: Angiogenesis; Corneal Keratocyte; Cytokines; Inflammation; Microarray Analysis; OLETF