Wang Xue-Feng, Cui Jing Z, Nie Weijia, Prasad Shiv S, Matsubara Joanne A
Department of Ophthalmology and Visual Sciences, The University of British Columbia, 2550 Willow Street, Vancouver, BC V5Z 3N9, Canada.
Exp Eye Res. 2004 Aug;79(2):209-21. doi: 10.1016/j.exer.2004.03.013.
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.
我们检测了通过反复传代在体外老化的视网膜色素上皮（RPE）细胞的基因表达谱。将来自人眼的RPE细胞培养至第3 - 5代（早期传代）或第19 - 21代（晚期传代），并用于通过cDNA微阵列研究基因表达谱。微阵列分析的结果通过实时PCR进一步证实。微阵列分析显示588个已知基因之间存在基因表达变化。使用两倍标准，15个基因（2.6%）在晚期传代RPE细胞中的表达水平升高，而43个基因（7.3%）的表达水平降低。这些差异表达的基因涵盖了许多功能类别。少数应激基因，如簇集蛋白、复制蛋白A和Ku80，上调。下调的基因包括许多能量和生物分子代谢酶以及细胞周期蛋白和细胞粘附蛋白。实时PCR的结果与微阵列结果总体一致。所检测的血管生成因子的表达水平要么未改变，要么下调。比较早期（第3 - 5代）和晚期（第9 - 12代）传代的RPE细胞，鉴定出几类差异表达的基因。然而，已知血管生成因子的表达并未增强。
