Identification of replicative senescence-associated genes in human umbilical vein endothelial cells by an annealing control primer system

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Title
Identification of replicative senescence-associated genes in human umbilical vein endothelial cells by an annealing control primer system
Author(s)
T W Kim; H J Kim; C H Lee; H Y Kim; S H Baek; J H Kim; Ki Sun Kwon; J R Kim
Bibliographic Citation
Experimental Gerontology, vol. 43, no. 4, pp. 286-295
Publication Year
2008
Abstract
Cellular senescence is regulated by specific genes in many organisms. The identification and functional analysis of senescence-associated genes could provide valuable insights into the senescence process. Here, we employed a new and improved differential display reverse transcription-polymerase chain reaction (DDRT-PCR) method that involves annealing control primers (ACPs) to identify genes that are differentially expressed in human umbilical endothelial cells during replicative senescence. Using 120 ACPs, we identified 31 differentially expressed genes (DEGs). Basic local alignment search tool (BLAST) search revealed 29 known genes and two unknown genes. Expression levels of the 29 known genes were confirmed by real-time quantitative RT-RCR and by Western blotting for eight of these genes. CD9 antigen, MHC class I chain-related sequence A (MICA) and cell division cycle 37 homolog (CDC37) were up-regulated, and bone morphogenetic protein 4 (BMP4), dickkopf-1 (DKK1), and transcription factor 7-like 1 (TCF7L1) were down-regulated in old cells. Treatment with recombinant human MICA caused a decrease in cell proliferation and an increase in senescence-associated β-galactosidase staining. Further analysis of differentially expressed genes may provide insights into the molecular basis of replicative senescence and vascular diseases associated with cellular senescence.
Keyword
Cell agingDDRT-PCREndothelial cellsGene expression
ISSN
0531-5565
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.exger.2007.12.010
Type
Article
Appears in Collections:
Division of Research on National Challenges > Aging Research Center > 1. Journal Articles
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