Identification of replicative senescence-associated genes in human umbilical vein endothelial cells by an annealing control primer system.
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 beta-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.
Kim
Tae Woo
TW
Department of Biochemistry and Molecular Biology, College of Medicine, Yeungnam University, Republic of Korea.
Kim
Hyun Jung
HJ
Lee
Chuhee
C
Kim
Hwa Young
HY
Baek
Suk-Hwan
SH
Kim
Jung Hye
JH
Kwon
Ki-Sun
KS
Kim
Jae-Ryong
JR
D015703:Antigens, CD / Q000378:metabolism; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D016922:Cellular Senescence / Q000235:genetics*; D017931:DNA Primers; D015536:Down-Regulation; D042783:Endothelial Cells / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods*; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D018832:Molecular Chaperones / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D051820:TCF Transcription Factors / Q000378:metabolism; D060245:Tetraspanin 29; D058125:Transcription Factor 7-Like 1 Protein; D014471:Umbilical Veins / Q000378:metabolism*; D015854:Up-Regulation
D015703:Antigens, CD; C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins; C554869:CD9 protein, human; C501510:CDC37L1 protein, human; D018797:Cell Cycle Proteins; C501126:DKK1 protein, human; D017931:DNA Primers; D015395:Histocompatibility Antigens Class I; D036341:Intercellular Signaling Peptides and Proteins; C099187:MHC class I-related chain A; D008562:Membrane Glycoproteins; D018832:Molecular Chaperones; D051820:TCF Transcription Factors; C544912:TCF7L1 protein, human; D060245:Tetraspanin 29; D058125:Transcription Factor 7-Like 1 Protein