Differential gene expression of early and late passage retinal pigment epithelial cells.
We examined the gene expression profiles of retinal pigment epithelial (RPE) cells which were aged in vitro by repeated passage. RPE cells from human eyes were cultured to passage 3-5 (early passage) or 19-21 (late passage) and used to study gene expression profiles by cDNA microarray. Results from microarray analysis were further confirmed by real-time PCR. Microarray analysis showed gene expression changes among 588 known genes. The expression levels of 15 genes (2.6%) increased in late passage RPE cells, while 43 genes (7.3%) decreased using a two-fold criterion. These differentially expressed genes encompassed many functional classes. A small number of stress genes, such as clusterin, replication protein A and Ku80, were up-regulated. The down-regulated genes included many enzymes of energy and biomolecule metabolism as well as cell cycle proteins and cell adhesion proteins. Results from real-time PCR were generally consistent with microarray findings. The expression levels of the examined angiogenic factors were either unchanged or down-regulated. Comparing early (p=3-5) and late (p=9-12) passage RPE cells, several categories of differentially expressed genes were identified. However, there was no enhanced expression of known angiogenic factors.
Department of Ophthalmology and Visual Sciences, The University of British Columbia, 2550 Willow Street, Vancouver, BC V5Z 3N9, Canada.
Xue-Feng
XF
Wang
Jing Z
JZ
Cui
Weijia
W
Nie
Shiv S
SS
Prasad
Joanne A
JA
Matsubara
D043925:Angiogenesis Inducing Agents / Q000378:metabolism; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D006801:Humans; D008268:Macular Degeneration / Q000378:metabolism* / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D001616:beta-Galactosidase / Q000378:metabolism
D043925:Angiogenesis Inducing Agents; D001616:beta-Galactosidase