| Detection of cell cycle- and differentiation stage-dependent human telomerase reverse transcriptase expression in single living cancer cells. | |
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MedLine Citation:
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PMID: 16584924 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Elevated telomerase activity is an important molecular signature of cancer cells and primitive cells in regenerative tissues. However, isolation of single living cells with endogenous telomerase activity has not yet been possible. Here, we developed adenovirus serotype 35 tropism-based vectors encoding destabilized enhanced green fluorescence protein with a half-life of 2 h (d2EGFP) driven by the human telomerase reverse transcriptase (hTERT) promoter. As assessed in telomerase-positive or -negative cell lines, the d2EGFP expression positively correlated with hTERT transcript content and telomerase activity. In retinoic acid-induced differentiating HL-60 cells, the d2EGFP expression is diminished in the same manner as the hTERT expression. Individual cells from HeLa and HL-60 cell lines exhibited heterogeneous d2EGFP expression, which was cell cycle dependent, as the sorted d2EGFP+ HL-60 cells contained twice as many cells in S/G2/M phase of the cell cycle compared with the d2EGFP- HL-60 cells. However, both cell populations exhibited the same proliferation and regeneration capacities. Heterogeneous d2EGFP expression was also detected in xenograft glioblastoma multiforme cells with tumor formation capacity. Thus, d2EGFP expression reported cell cycle- and differentiation stage-dependent hTERT expression. Our study facilitates isolation and characterization of single living cells with telomerase activity. |
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Authors:
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Anna Edqvist; Johan Rebetz; Marcus Järås; Anna Rydelius; Gunnar Skagerberg; Leif G Salford; Bengt Widegren; Xiaolong Fan |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2006-04-11 |
Journal Detail:
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Title: Molecular therapy : the journal of the American Society of Gene Therapy Volume: 14 ISSN: 1525-0016 ISO Abbreviation: Mol. Ther. Publication Date: 2006 Jul |
Date Detail:
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Created Date: 2006-06-23 Completed Date: 2006-09-05 Revised Date: 2008-11-21 |
Medline Journal Info:
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Nlm Unique ID: 100890581 Medline TA: Mol Ther Country: United States |
Other Details:
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Languages: eng Pagination: 139-48 Citation Subset: IM |
Affiliation:
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Section of Immunology, Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, Lund University, Lund, Sweden |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Adenoviridae
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genetics Animals Cell Cycle / drug effects, physiology* Cell Differentiation / drug effects, physiology* Cell Line Cell Line, Tumor Cells, Cultured DNA-Binding Proteins / genetics*, metabolism Gene Expression Regulation, Enzymologic Gene Therapy / methods Genetic Vectors / genetics Green Fluorescent Proteins / genetics, metabolism HL-60 Cells Hela Cells Humans K562 Cells Mice Mice, SCID Models, Genetic Neoplasms / genetics*, pathology, therapy Promoter Regions, Genetic / genetics Reverse Transcriptase Polymerase Chain Reaction Telomerase / genetics*, metabolism Tretinoin / pharmacology Xenograft Model Antitumor Assays |
| Chemical | |
Reg. No./Substance:
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0/DNA-Binding Proteins; 147336-22-9/Green Fluorescent Proteins; 302-79-4/Tretinoin; EC 2.7.7.49/Telomerase |
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