Evaluation of differentially expressed genes identified in keratoconus.
OBJECTIVE
To identify the differentially expressed genes (DEGs) in the human keratocytes in keratoconus.


METHODS
Total RNA extracted from cultured corneal stromal fibroblasts from normal and keratoconic corneas were used for the synthesis of cDNA. DEGs were screened by an annealing control primer(TM)-based PCR method using GeneFishing() DEG kits. The differentially expressed bands were sequenced and analyzed. The genes identified were further evaluated by reverse transcriptase PCR and quantitative real-time PCR.


RESULTS
Overexpression of bone morphogenetic protein 4 (BMP4), cofilin 1 (CFL1), and JAW1-related protein (MRVI1) and underexpression of actin, alpha 2 (ACTA2), gene rich cluster, and C 10 gene (GRCC10), tissue inhibitor of metalloproteinase 3 (TIMP3), tissue inhibitor of metalloproteinase 1 (TIMP1), and somatostatin receptor 1 (SSTR1) were verified, and these results were confirmed by reverse transcriptase PCR and quantitative real-time PCR.


CONCLUSIONS
Eight genes were identified to be differentially expressed in keratoconus and related with apoptosis, the cytoskeleton, wound healing, and nerve fibers. The genes identified may be involved in the mechanism underlying stromal thinning; thus, they could be important and deserve further investigation.
Lee
Ji-Eun
JE
The Department of Ophthalmology, College of Medicine, Pusan National University, Pusan, Korea.
Oum
Boo Sup
BS
Choi
Hee Young
HY
Lee
Seung Uk
SU
Lee
Jong Soo
JS
D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D007640:Keratoconus / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D005136:Eye Proteins