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G. J. S. Jenkins C. Morgan J. N. Baxter E. M. Parry J. M. Parry 《Mutation Research - Genetic Toxicology and Environmental Mutagenesis》2001,498(1-2)
We have analysed five mutation hotspots within the p53 gene (codons 175, 213, 248, 249, and 282) for mutations induced by hydrogen peroxide (H2O2), employing the restriction site mutation (RSM) assay. In addition, four other restriction sites covering non-hotspot codons of exons 5–9 of the p53 gene (codons 126, 153/54, 189 and the 3′ splice site of exon 9) were analysed by the RSM assay for H2O2-induced mutations. Two cell types were concurrently analysed in this study, i.e. primary fibroblast cells and a gastric cancer cell line. Using the RSM assay, H2O2-induced mutations were only detected in exon 7 of the p53 gene. This was true for both cell types. These mutations were mainly induced in the Msp I restriction site (codon 247/248) and were predominantly GC to AT transitions (71%). Hence these GC to AT mutations were presumably due to H2O2 exposure, possibly implicating the 5OHdC adduct, which is known to induce C to T mutations upon misreplication. Importantly, this study demonstrates that the RSM methodology is capable of detecting rare oxidative mutations within the hotspot codons of the p53 tumour suppressor gene. Hence, this methodology may allow the detection of early p53 mutations in pre-malignant tissues. 相似文献
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The effects of 1,6-dinitropyrene (1,6-DNP) on the fidelity of cell division were studied in the transformed human fibroblast cell line MRC5VA. Over a dose range of 0.1-10 micrograms/ml of 1.6-DNP, we observed significant increases in the levels of abnormal division stages, associated with damage to the spindle apparatus of the cell. Qualitative changes in spindle morphology and a quantitative decrease in pole-to-pole spindle length were also observed with increasing doses of 1.6-DNP. Such changes in the size and morphology of the spindle corresponded with an accumulation of cells blocked at metaphase. The presence of catalase did not modify the response, suggesting that the effects on the spindle apparatus and cell division were not caused by the generation of radicals but by the direct action of 1.6-DNP. 相似文献
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Culturing satellite cells from living single muscle fiber explants 总被引:21,自引:0,他引:21
J. David Rosenblatt Alison I. Lunt David J. Parry Terence A. Partridge 《In vitro cellular & developmental biology. Animal》1995,31(10):773-779
Summary Conventional methods for isolating myogenic (satellite) cells are inadequate when only small quantities of muscle, the tissue
in which satellite cells reside, are available. We have developed a tissue culture system that reliably permits isolation
of intact, living, single muscle fibers with associated satellite cells from predominantly fast and slow muscles of rat and
mouse; maintenance of the isolated fibers in vitro; dissociation, proliferation, and differentiation of satellite cells from
each fiber; and removal of the fiber from culture for analysis. 相似文献