Microarray analysis of replicative senescence

Abstract

Replicative senescence triggers mRNA expression patterns that vary widely and cell lineage strongly influences these patterns. In fibroblasts, the senescent state mimics inflammatory wound repair processes and, as such, senescent cells may contribute to chronic wound pathologies.

Exploration of replicative senescence-associated genes in human dermal fibroblasts by cDNA microarray technology

“…We then validated the cDNA microarray analysis using semi-quantitative RT-PCR and Western blot analysis to reduce the number of false positives. Although our results are generally in line with those of previous studies (Shelton et al, 1999;Ly et al, 2000;Zhang et al, 2003), we observed alterations in the expression of many novel genes in replicative-senescent fibroblasts.…”

“…3) were consistent by cDNA microarray and RT-PCR. Groa, IGF-binding protein 5 (IGFBP5) (Shelton et al, 1999), interferon a-inducible protein (GIP3) (Tahara et al, 1995), IL-6 (Goodman and Stein, 1994), proenkephalin (Caffrey et al, 1994), and BCG-induced gene in monocytes (BIGM103) have already been reported to be up-regulated. And, osteoblast specific factor 2 (OSF2), centromeric protein F (CENP-F), fibrillin 2, cyclooxygenease 2 (Ly et al, 2000), elastin (Shelton et al, 1999), and meningioma (MN1) were found to be down-regulated in aged human fibroblasts.…”

“…Groa, IGF-binding protein 5 (IGFBP5) (Shelton et al, 1999), interferon a-inducible protein (GIP3) (Tahara et al, 1995), IL-6 (Goodman and Stein, 1994), proenkephalin (Caffrey et al, 1994), and BCG-induced gene in monocytes (BIGM103) have already been reported to be up-regulated. And, osteoblast specific factor 2 (OSF2), centromeric protein F (CENP-F), fibrillin 2, cyclooxygenease 2 (Ly et al, 2000), elastin (Shelton et al, 1999), and meningioma (MN1) were found to be down-regulated in aged human fibroblasts. However, the following were found for the first time to be associated with senescence; CD36, putative lymphocyte G 0 / G 1 switch gene (G 0 S 2 ), hexokinase 2, tumor protein D52-like 1 (TPD52L1), erythrocyte membrane protein band 4.1-like 3 (EPB41L3/Dal-1), myxovirus resistant gene (MX1), dipeptidylpeptidase 4 (DPP4, CD26), B-factor, chemokine (C-X-C motif) ligand 6 (CXCL6), phospholipase A2 receptor 1 (PLA2R1), immunoglobulin superfamily containing leucine-rich repeat (ISLR), neurotrimin (HNT), dematopontin (DMP), kinesin family member 4A (KIF4A), IGF2-associated protein (IGF2A), apoptosis-related RNA binding protein (NAPOR3), and RDC1 homolog.…”

“…RNA differential displays and the serial assessment of gene expression (Welle et al, 2001) have been applied to explore senescence-associated genes to gain an insight into the molecular mechanisms underlying senescence. Moreover, recent advances in cDNA array technology have made it possible to analyze global gene expressions, and a variety of genes involved in cell cycle regulation, immune and inflammation, cytoskeleton, stress response, and metabolism are known to be altered during cellular senescence (Shelton et al, 1999;Ly et al, 2000;Zhang et al, 2003) and animal models (Lee et al, 1999(Lee et al, , 2000Tollet-Egnell et al, 2001;Kayo et al, 2001;Cao et al, 2001). Shelton et al (1999) assessed senescence-associated gene expression in dermal fibroblasts using a 1 k cDNA microarray and concluded that the senescent state mimics inflammatory wound repair processes.…”

DBC1 re-expression alters the expression of multiple components of the plasminogen pathway

“…We detected the 47 kDa nonglycosylated (intracellular, lysates) and the 60-70 kDa glycosylated (secreted) forms of SERPINB5 using Western blotting (Figure 4, panel c). SERPINB5 has been reported as an upregulated gene in senescent dermal fibroblasts cells (Shelton et al, 1999) along with MT-1E, PLAU, PLAT, IL1B, IGFBP3 and superoxide dismutase (SOD2), which were all differentially expressed in our experiments (Table 1). Similarly, MT-2 and TFPI-2 (2.2-fold upregulated in DBC1 clone B) were identified as overexpressed during ras-induced senescence (Barradas et al, 2002).…”