کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
7233045 1470970 2014 38 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A cell-based biosensor system HepG2CDKN1A-DsRed for rapid and simple detection of genotoxic agents
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
A cell-based biosensor system HepG2CDKN1A-DsRed for rapid and simple detection of genotoxic agents
چکیده انگلیسی
The regulatory requirements for genotoxicity testing rely on a battery of genotoxicity tests, which generally consist of bacterial and mammalian cell assays for detection of gene mutations and chromosomal aberrations. However, for rapid screening, these methods are not appropriate. We have developed a new cell-based biosensor system that provides rapid and simple detection of genotoxic substances. This is based on stable transfection of human hepatoma HepG2 cells with a plasmid that encodes the red fluorescent protein DsRed2 under the control of the CDKN1A promoter (HepG2CDKN1A-DsRed cells). As the major downstream target gene of activated TP53, the tumour-suppressor gene CDKN1A is responsible for cell-cycle arrest following DNA damage, and it has been shown to be specifically up-regulated by genotoxic carcinogens. The assay is optimised for a 96-well microplate format and spectrofluorimetric quantification of induced DsRed expression. The assay was evaluated by testing direct-acting and indirect-acting genotoxic compounds with different mechanisms of action, along with non-genotoxic compounds. Out of 25 compounds that are known to be genotoxic in vitro and in vivo, 21 (84%) are detected as positive at non-cytotoxic doses, whereas of 12 compounds not considered genotoxic, 11 (92%) are negative. These data indicate the high sensitivity and specificity of our biosensor system. Based on its simplicity and sensitivity, this biosensor developed with HepG2CDKN1A-DsRed cells has the potential to become a valuable tool for genotoxicity screening for chemical safety evaluation, as well as for environmental and occupational monitoring of exposure to genotoxic agents and their complex mixtures.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biosensors and Bioelectronics - Volume 61, 15 November 2014, Pages 102-111
نویسندگان
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