The upregulation of angiogenic gene expression in cultured retinal pigment epithelial cells grown on type I collagen.
OBJECTIVE
Increases in matrix proteins, such as type I collagen and fibronectin, are observed with aging in the retinal pigment epithelium (RPE) basement membrane. However, little is known about altered gene expression profiles of RPE associated with increases in matrix proteins. We investigated changes in gene expression profiles of a human RPE cell line (ARPE-19) cultured on type I collagen.


METHODS
Visually confluent ARPE-19 cells were grown on either Matrigel (M group) or type I collagen (C group) without serum over 3 days. Total RNA was extracted and reverse transcribed. Gene expression profiles in both groups were compared using microarray analyses. Several angiogenic genes including integrin alpha V, integrin alpha 2, integrin beta 1, integrin beta 3, VEGF-A, VEGF-B, and VEGF-C were subjected to quantitative analyses using real-time PCR.


RESULTS
Out of 192 genes examined, angiogenesis-related genes (17.7%) and extracellular matrix-related genes (30.2%) were expressed highly (with more than 1.5-fold difference) in the C group when compared with the M group. In real-time PCR analyses, all VEGF and integrin family genes examined were expressed more in the C group than in the M group.


CONCLUSIONS
Type I collagen likely causes an upregulation in a number of angiogenic gene expression patterns as seen in RPE in vitro experiments.
Imai
Hisanori
H
Department of Surgery, Division of Ophthalmology, Kobe University Graduate School of Medicine, Chuo-ku, Kobe, Japan.
Honda
Shigeru
S
Kondo
Naoshi
N
Ishibashi
Kazuki
K
Tsukahara
Yasutomo
Y
Negi
Akira
A
D002460:Cell Line; D024042:Collagen Type I / Q000494:pharmacology*; D003199:Computer Systems; D003470:Culture Media / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D038982:Integrin alpha2beta1 / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D015854:Up-Regulation / Q000187:drug effects*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D024042:Collagen Type I; D003470:Culture Media; D038982:Integrin alpha2beta1; D042461:Vascular Endothelial Growth Factor A