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標題: The limitations and validities of senescence associated-beta-galactosidase activity as an aging marker for human foreskin fibroblast Hs68 cells
作者: Yang, N.C.
Hu, M.L.
關鍵字: aging marker;senescence associated-beta-galactosidase;lysosomal;galactosidase;fluorescein di-beta-D-galactopyranoside;reliability;specificity;human-diploid fibroblasts;replicative senescence;in-vivo;biomarkers
Project: Experimental Gerontology
期刊/報告no:: Experimental Gerontology, Volume 40, Issue 10, Page(s) 813-819.
The senescence associated-beta-galactosidase (SA-G) 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-PG 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-beta G 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-beta G 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 beta G 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-beta G activity. These results support the contention that SA-beta G is the lysosomal beta G 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-beta G activity, rendering SA-beta G activity unspecific for cell-aging. Thus, the assay for cell-aging is only reliable when these confounding factors are controlled or excluded. (c) 2005 Elsevier Inc. All rights reserved.
ISSN: 0531-5565
DOI: 10.1016/j.exger.2005.07.011
Appears in Collections:食品暨應用生物科技學系

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