Document Detail


Characterization of the activities of p21Cip1/Waf1 promoter-driven reporter systems during camptothecin-induced senescence-like state of BHK-21 cells.
MedLine Citation:
PMID:  16779499     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
It was attempted in this work to establish a cell line in which senescent cells can be readily and directly identified in situ in live culture. Transcriptional activation of p21(Cip1/Waf1) gene is known to be one of the key steps in the development of cellular senescence, whereas the elements within the p21(Cip1/Waf1) promoter that regulate the transcriptional activation of p21(Cip1/Waf1) during cellular senescence have not been clearly defined. Thus, several reporter plasmids were constructed in each of which the gene of green fluorescent protein was placed under the control of a selected fragment of p21(Cip1/Waf1) promoter, and stably transfected into BHK-21 cells. The transfected cells were induced to become senescence-like by camptothecin and assayed for fluorescence intensity. It was shown that the reporter system constructed with bases -2504 to +406 of the p21(Cip1/Waf1) promoter was very efficient in reflecting the senescence of BHK-21 cells by increased cytosolic fluorescence, and the fluorescence intensity of senescent cells was easily distinguished from that of quiescent cells.
Authors:
Hsueh-Ling Cheng; Shiou-Ming Chang; Ya-Wen Cheng; Hung-Jen Liu; Yo-Chia Chen
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-05-24
Journal Detail:
Title:  Molecular and cellular biochemistry     Volume:  291     ISSN:  0300-8177     ISO Abbreviation:  Mol. Cell. Biochem.     Publication Date:  2006 Oct 
Date Detail:
Created Date:  2006-10-12     Completed Date:  2006-12-19     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0364456     Medline TA:  Mol Cell Biochem     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  29-38     Citation Subset:  IM    
Affiliation:
Institute of Biotechnology, National Pingtung University of Science and Technology, Taiwan, ROC.
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MeSH Terms
Descriptor/Qualifier:
Animals
Apoptosis / drug effects
Camptothecin / pharmacology*
Cell Aging / drug effects*
Cell Cycle / drug effects
Cricetinae
Cyclin-Dependent Kinase Inhibitor p21 / genetics*
Flow Cytometry
Fluorescence
Genes, Reporter*
Green Fluorescent Proteins / metabolism
Humans
Plasmids / metabolism
Promoter Regions, Genetic / genetics*
Transfection
beta-Galactosidase / metabolism
Chemical
Reg. No./Substance:
0/CDKN1A protein, human; 0/Cyclin-Dependent Kinase Inhibitor p21; 147336-22-9/Green Fluorescent Proteins; 7689-03-4/Camptothecin; EC 3.2.1.23/beta-Galactosidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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