Document Detail


The limitations and validities of senescence associated-beta-galactosidase activity as an aging marker for human foreskin fibroblast Hs68 cells.
MedLine Citation:
PMID:  16154306     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The senescence associated-beta-galactosidase (SA-betaG) assay has become one of the most commonly used markers of cell-aging. However, the reliability of the assay is questionable because the enzyme is a non-specific marker for cell-aging. In this study, we found that the SA-betaG activity increased with cell age as well as in confluent quiescent cells or cells under serum starvation, and in cells treated with H2O2. Importantly, we found that SA-betaG activity was irreversibly increased in the senescent cells or H2O2-teated cells, but was reversible in quiescent cells induced by serum starvation or confluence. Using fluorescein di-beta-d-galactopyranoside (FDG) method for SA-betaG detection, we showed that senescent human foreskin fibroblast Hs68 cells did not express a specific enzyme that has a maximal activity at pH 6.0. In the pH profile of the cellular betaG activity in senescent Hs68 cells, only a single peak was found (with maximum at pH 4.6), and no addition peak was found at or around pH 6.0 that could be attributed to the SA-betaG activity. These results support the contention that SA-betaG is the lysosomal betaG that is detectable at suboptimal pH (i.e. pH 6.0) and demonstrate that cell-aging is not the only factor that can increase SA-betaG activity, rendering SA-betaG activity unspecific for cell-aging. Thus, the assay for cell-aging is only reliable when these confounding factors are controlled or excluded.
Authors:
Nae-Cherng Yang; Miao-Lin Hu
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2005-09-08
Journal Detail:
Title:  Experimental gerontology     Volume:  40     ISSN:  0531-5565     ISO Abbreviation:  Exp. Gerontol.     Publication Date:  2005 Oct 
Date Detail:
Created Date:  2005-10-10     Completed Date:  2006-04-03     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0047061     Medline TA:  Exp Gerontol     Country:  England    
Other Details:
Languages:  eng     Pagination:  813-9     Citation Subset:  IM    
Affiliation:
Department of Biotechnology, Chungchou Institute of Technology, Changhua, Taiwan, ROC.
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MeSH Terms
Descriptor/Qualifier:
Aged
Biological Markers / analysis
Cell Aging / physiology*
Cell Line
Fibroblasts / enzymology*
Histocytochemistry / methods
Humans
Male
Models, Biological
Skin
beta-Galactosidase / analysis*
Chemical
Reg. No./Substance:
0/Biological Markers; EC 3.2.1.23/beta-Galactosidase

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