Induction of replicative senescence biomarkers by sublethal oxidative stresses in normal human fibroblast.
We tested the long-term effects of sublethal oxidative stresses on replicative senescence. WI-38 human diploid fibroblasts (HDFs) at early cumulative population doublings (CPDs) were exposed to five stresses with 30 microM tert-butylhydroperoxide (t-BHP). After at least 2 d of recovery, the cells developed biomarkers of replicative senescence: loss of replicative potential, increase in senescence-associated beta-galactosidase activity, overexpression of p21(Waf-1/SDI-1/Cip1), and inability to hyperphosphorylate pRb. The level of mRNAs overexpressed in senescent WI-38 or IMR-90 HDFs increased after five stresses with 30 microM t-BHP or a single stress under 450 microM H(2)O(2). These corresponding genes include fibronectin, osteonectin, alpha1(I)-procollagen, apolipoprotein J, SM22, SS9, and GTP-alpha binding protein. The common 4977 bp mitochondrial DNA deletion was detected in WI-38 HDFs at late CPDs and at early CPDs after t-BHP stresses. In conclusion, sublethal oxidative stresses lead HDFs to a state close to replicative senescence.
Dumont
P
P
The University of Namur (FUNDP), Department of Biology, Laboratory of Cellular Biochemistry and Biology, Namur, Belgium.
Burton
M
M
Chen
Q M
QM
Gonos
E S
ES
Frippiat
C
C
Mazarati
J B
JB
Eliaers
F
F
Remacle
J
J
Toussaint
O
O
D015415:Biomarkers / Q000032:analysis; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D016922:Cellular Senescence / Q000502:physiology*; D051152:Clusterin; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000235:genetics; D004791:Enzyme Inhibitors / Q000032:analysis; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D005353:Fibronectins / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D006023:Glycoproteins / Q000235:genetics; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D018832:Molecular Chaperones; D015676:Osteonectin / Q000235:genetics; D018384:Oxidative Stress / Q000502:physiology*; D011347:Procollagen / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics; D020122:tert-Butylhydroperoxide / Q000494:pharmacology
D015415:Biomarkers; C493956:CDKN1A protein, human; C495242:CLU protein, human; D051152:Clusterin; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D004791:Enzyme Inhibitors; D005353:Fibronectins; D006023:Glycoproteins; D018832:Molecular Chaperones; D015676:Osteonectin; D011347:Procollagen; D020122:tert-Butylhydroperoxide; D006861:Hydrogen Peroxide; D001616:beta-Galactosidase