open access

Vol 50, No 2 (2012)
ORIGINAL PAPERS
Submitted: 2012-07-04
Accepted: 2012-07-04
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The profile of lysosomal exoglycosidases in replicative and stress-induced senescence in early passage human fibroblasts

Małgorzata Knaś, Anna Zalewska, Rafał Krętowski, Marek Niczyporuk, Napoleon Waszkiewicz, Marzanna Cechowska-Pasko, Danuta Waszkiel, Krzysztof Zwierz
DOI: 10.5603/FHC.2012.0031
·
Folia Histochem Cytobiol 2012;50(2):220-227.

open access

Vol 50, No 2 (2012)
ORIGINAL PAPERS
Submitted: 2012-07-04
Accepted: 2012-07-04

Abstract

The aim of the present study was to assess the profiles of the exoglycosidases: N-acetyl-β-hexosoaminidase, β glucuronidase and β galactosidase, α mannosidase and α fucosidase in fibroblast culture undergoing replicative and stress-induced senescence. Half of the cell culture was grown in normal conditions, without the stressor, and the other half of the cell was treated with 0.15 mM tert-butylhydroperoxide. The activities of total N-acetyl-β-hexosoaminidase as well as β glucuronidase in the cell lysate were determined in duplicates using the method of Marciniak et al. The activities of β galactosidase, α mannosidase and α fucosidase in the cell lysate were determined in duplicates using the method of Chatteriee et al. with the modification by Zwierz et al. The activities of the exoglycosidases examined, with the exception of β glucuronidase, showed a significant increase between individual days of the experiment in both non-stressed and stressed fibroblast cell culture. On each day of the experiment, in the cell lysate of stressed fibroblasts, the activities of exoglycosidases were significantly higher compared to the non-stressed cells. There were very strong correlations between SA-β-GAL staining and b galactosidase activity on individual days of the experiment in both non-stressed and stressed fibroblast cell culture. Replicative and stress-induced senescence results in significant changes to the level of lysosomal exoglycosidases, and results in enhanced lysosomal degradative capacity.

Abstract

The aim of the present study was to assess the profiles of the exoglycosidases: N-acetyl-β-hexosoaminidase, β glucuronidase and β galactosidase, α mannosidase and α fucosidase in fibroblast culture undergoing replicative and stress-induced senescence. Half of the cell culture was grown in normal conditions, without the stressor, and the other half of the cell was treated with 0.15 mM tert-butylhydroperoxide. The activities of total N-acetyl-β-hexosoaminidase as well as β glucuronidase in the cell lysate were determined in duplicates using the method of Marciniak et al. The activities of β galactosidase, α mannosidase and α fucosidase in the cell lysate were determined in duplicates using the method of Chatteriee et al. with the modification by Zwierz et al. The activities of the exoglycosidases examined, with the exception of β glucuronidase, showed a significant increase between individual days of the experiment in both non-stressed and stressed fibroblast cell culture. On each day of the experiment, in the cell lysate of stressed fibroblasts, the activities of exoglycosidases were significantly higher compared to the non-stressed cells. There were very strong correlations between SA-β-GAL staining and b galactosidase activity on individual days of the experiment in both non-stressed and stressed fibroblast cell culture. Replicative and stress-induced senescence results in significant changes to the level of lysosomal exoglycosidases, and results in enhanced lysosomal degradative capacity.
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Keywords

exoglycosidases; fibroblast cell culture; replicative and stress-induced senescence

About this article
Title

The profile of lysosomal exoglycosidases in replicative and stress-induced senescence in early passage human fibroblasts

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 50, No 2 (2012)

Pages

220-227

DOI

10.5603/FHC.2012.0031

Bibliographic record

Folia Histochem Cytobiol 2012;50(2):220-227.

Keywords

exoglycosidases
fibroblast cell culture
replicative and stress-induced senescence

Authors

Małgorzata Knaś
Anna Zalewska
Rafał Krętowski
Marek Niczyporuk
Napoleon Waszkiewicz
Marzanna Cechowska-Pasko
Danuta Waszkiel
Krzysztof Zwierz

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