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Abstract
The isoflavone irilone is found in human plasma after ingestion of red clover-based dietary supplements, but information allowing safety assessment is rare. Here, data in support of the mutagenic potential of irilone in cultured V79 cells [1] are presented. These data include (i) a quantitative assessment of irilone in the culture medium during the cell culture experiments, (ii) changes in the mutation spectrum in cDNA of the hypoxanthine-guanine phosphoribosyltransferase locus of irilone-treated V79 cells, (iii) occurrence of karyorrhexis and apoptosis as well as (iv) number of micronucleated cells containing whole chromosomes or chromosomal fragments. Also exemplary micrographs, used for the fluorescence microscopic assessment of (iii) and (iv) are presented.
Highlights
The isoflavone irilone is found in human plasma after ingestion of red clover-based dietary supplements, but information allowing safety assessment is rare
Mutation spectrum in cDNA of irilonefeatures treated V79 cells was determined by Sanger method
Karyorrhexis, and micronucleus formation were assessed by fluorescence microscopy
Summary
The mutation spectra of IRI-induced mutants formed during the hypoxanthine-guanine phosphoribosyltransferase (HPRT) assay in cultured V79 cells was investigated. Exon deletions at cDNA level are caused by both deletions, base pair substitutions, and frameshift mutations at DNA level [5], the increase in exon deletions in mutation spectra of 6-TG resistant V79 cells indicated a clastogenic mode of action, which is known to be detected insufficiently by the Hprt gene mutation assay [10]. After treatment with the known aneugen diethylstilbestrol (15 and 25 mM), the frequency of cells with micronuclei containing whole chromosomes at 0–15 h postincubation time was increased to maximum 89 micronucleated cells/3000 cells (25 mM, 6 h postincubation) compared to 2.6–6.1 micronucleated cells/3000 cells observed with the solvent control (Supplemental Table, and [1]). 3000 cells (Supplemental Table, and [1])
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