Induction of senescence-associated genes by 5-bromodeoxyuridine in HeLa cells.
5-Bromodeoxyuridine (BrdU) universally induces a senescence-like phenomenon in mammalian cells. To assess this phenomenon at the level of gene expression, we constructed a PCR-based subtractive cDNA library enriched for mRNA species that immediately increase by administration of BrdU to HeLa cells. Candidate cDNA clones were isolated by differential colony hybridization, and then positive clones were identified by Northern blot analysis. Sequencing analysis revealed that the identified cDNA species were classified into three groups: widely used senescence-markers, known species whose relevance to senescence is yet to be reported, and known or novel ESTs. As expected, the majority of them showed an increase in expression in senescent human diploid fibroblasts. These results suggest that similar mechanisms operate in the regulation of BrdU-induced genes and senescence-associated genes.
Kihara Institute of Biological Research and Graduate School of Integrated Science, Yokohama City University, Maioka-cho 641-12, Totsuka-ku, 244-0813, Yokohama, Japan.
T
T
Suzuki
S
S
Minagawa
E
E
Michishita
H
H
Ogino
M
M
Fujii
Y
Y
Mitsui
D
D
Ayusawa
D001973:Bromodeoxyuridine / Q000494:pharmacology*; D016922:Cellular Senescence / Q000235:genetics*; D017364:Collagenases / Q000235:genetics; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000235:genetics; D020224:Expressed Sequence Tags; D005353:Fibronectins / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects*; D015723:Gene Library; D006367:HeLa Cells; D006801:Humans; D016133:Polymerase Chain Reaction; D014158:Transcription, Genetic / Q000187:drug effects
C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D005353:Fibronectins; D017364:Collagenases; C417249:collagenase 1; D001973:Bromodeoxyuridine