Microarray Analysis for Genes Associated with Angiogenesis in Diabetic OLETF Keratocytes.
10.3346/jkms.2014.29.2.265
- Author:
Jun Mo PARK
1
;
Young Min PARK
;
Wook JUNG
;
Ji Eun LEE
;
Jong Soo LEE
Author Information
1. Department of Ophthalmology, Busan St. Mary's Hospital, Busan, Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Angiogenesis;
Cytokines;
Inflammation;
Corneal Keratocyte;
Microarray Analysis;
OLETF
- MeSH:
Animals;
Cells, Cultured;
Gene Expression Regulation/drug effects;
Interleukin-1alpha/pharmacology;
Keratinocytes/cytology/drug effects/*metabolism;
Neovascularization, Physiologic/*genetics;
*Oligonucleotide Array Sequence Analysis;
Rats;
Real-Time Polymerase Chain Reaction;
Receptor, Angiotensin, Type 1/genetics/metabolism;
Tumor Necrosis Factor-alpha/pharmacology
- From:Journal of Korean Medical Science
2014;29(2):265-271
- CountryRepublic of Korea
- Language:English
-
Abstract:
The purpose of this study was to identify the differences in angiogenesis gene expression between normal and diabetic keratocytes stimulated with interleukin-1alpha (IL-1alpha) and tumor necrosis factor-alpha (TNF-alpha). Primarily cultured normal and diabetic keratocytes were treated with 20 ng/mL of IL-1a and TNF-alpha 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-1alpha and TNF-alpha showed the newly expressed 15 and 14 genes, respectively. Newly and commonly under-expressed five genes followed by treatment with both IL-1alpha and TNF-alpha 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-alpha treatment. Il6 appeared strong expression after interleukin-1alpha treatment, but showed down expression after TNF-alpha treatment. Further studies to analyze and confirm the significance of the identified angiogenetic genes of diabetes are needed.
