Microarray for Genes Associated with Signal Transduction in Diabetic OLETF Keratocytes.
10.3341/kjo.2007.21.2.111
- Author:
Ji Eun LEE
1
;
Jong Soo LEE
;
Sang Ho HWANG
Author Information
1. Department of Ophthalmology, College of Medicine, Pusan National University, Pusan, Korea. jongsool@pusan.ac.kr
- Publication Type:Original Article
- Keywords:
Diabetes mellitus;
Keratocyte;
Microarray;
Rat;
Signaling transduction
- MeSH:
Animals;
Apoptosis;
Cells, Cultured;
Cornea/drug effects/*metabolism/pathology;
DNA/*genetics;
Diabetes Mellitus, Experimental/*genetics/pathology;
Gene Expression Profiling;
Insulin/genetics;
Interleukin-1alpha/pharmacology;
Mitogen-Activated Protein Kinase Kinases/genetics;
Nuclear Proteins/genetics;
Oligonucleotide Array Sequence Analysis/*methods;
Phosphoric Monoester Hydrolases/genetics;
Polymerase Chain Reaction;
Prolactin/genetics;
Rats;
Rats, Long-Evans;
Receptors, Notch/genetics;
Signal Transduction/drug effects/*genetics;
Transforming Growth Factor beta/genetics;
Tumor Necrosis Factor-alpha/pharmacology;
Ubiquitin-Protein Ligases/genetics
- From:Korean Journal of Ophthalmology
2007;21(2):111-119
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: The purpose of this study was to identify differences in signal transduction gene expression between normal and diabetic keratocytes stimulated with interleukin-1alpha (IL-1alpha) and tumor necrosis factor-alpha (TNF-alpha). METHODS: Normal and diabetic keratocytes were primarily cultured and treated with 20 ng/ml IL-1alpha and TNF-alpha for 6 h. cDNA was hybridized to an oligonucleotide microarray. Genes identified by the microarray were further evaluated by real-time PCR. RESULTS: Diabetic keratocytes over-expressed components of the MAPK and Notch pathways, and under-expressed components of the insulin, calcium, and TGF-beta pathways. Cytokine treated diabetic keratocytes differentially expressed components of the TGF-beta and MAPK pathways. After IL-1alpha and TNF-alpha treatment, nine genes were under-expressed, falling in the insulin, TGF-beta, and Toll-like receptor pathways. Real-time PCR showed a significant decrease in the IL-6 and TGF-beta2 genes and a significant increase in the Ppm1a gene. CONCLUSIONS: There were some differences in gene expression between normal and diabetic keratocytes related to signal transduction pathways, such as the insulin, MAPK, calcium, and TGF-beta pathways. In addition, IL-1alpha and TNF-alpha stimulating the insulin, TGF-beta, and Toll-like receptor signaling pathways may have different effects in diabetic keratocytes.
