Tandemly repeated DNA sequence instabilities induced by two promoters of morphological transformation in vitro: a short-term response to non-mutagenic agents in C3H/10T1/2 cells.

2.50
Hdl Handle:
http://hdl.handle.net/10146/68416
Title:
Tandemly repeated DNA sequence instabilities induced by two promoters of morphological transformation in vitro: a short-term response to non-mutagenic agents in C3H/10T1/2 cells.
Authors:
Parfett, Craig L.; Healy, Caroline
Abstract:
The ability of tumour promoters to alter DNA stability within regions that contain tandemly repeated sequences (TRSs), was studied in a cell culture model of multi-stage carcinogenesis. Non-cytotoxic concentrations of TPA (12-O-tetradecanoyl-phorbol-13-acetate) and xanthine oxidase with xanthine substrate, sufficient to promote morphological transformation in C3H/10T1/2 cultures, were tested for their effects on mutation frequencies in TRSs by a DNA fingerprinting approach. Specifically, restriction digests of genomic DNA samples from randomly selected, non-transformed clones, isolated from cultures after several days exposure to promoters, were visualized by Southern hybridizations with the multi-locus pentamer repeat sequence probe, Ms6-Hm (Pc-1). Basal and promoter-induced frequencies of sub-clone TRS fingerprint polymorphisms were estimated in five cell populations: an uncloned stock culture, three populations established from normal-appearing sub-clones, and one clonal population established from a 3-methylcholanthrene (MCA)-transformed focus. Basal variant fingerprint frequencies spanned a range from 0.0 to 0.43% mutants/band among cells from the four untransformed populations. Both TPA and xanthine oxidase treatments significantly increased recorded mutation frequencies, 2.3- and 2.7-fold, respectively, using combined data from the progenitor population and three untransformed clones. The untreated MCA-transformed clonal population appeared to contain a single, pre-existing mutant restriction fragment, but additional mutations were induced thereafter, in response to the promoting treatments. Taken together, the measured increases in mutations were highly significant and suggest that promoters of cell transformation in the C3H/10T1/2 cell line might induce a genome-wide instability targeted to regions containing Ms6-Hm sequence motifs.
Citation:
Mutat. Res. 2006, 604 (1-2):42-52
Journal:
Mutation research
Issue Date:
30-Apr-2006
URI:
http://hdl.handle.net/10146/68416
DOI:
10.1016/j.mrgentox.2005.12.008
PubMed ID:
16459133
Additional Links:
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T2D-4J625TV-1&_user=1843694&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000055040&_version=1&_urlVersion=0&_userid=1843694&md5=537d40f05453dcca95ca0161f896e2d1
Type:
Article
Language:
en
Description:
KEYWORDS - CLASSIFICATION: Animals;Base Sequence;chemistry;Canada;Carcinogens;Cell Culture Techniques;Cell Survival;Cells,Cultured;Chromosomal Instability;Clone Cells;drug effects;Dna;DNA Fingerprinting;DNA Primers;Environmental Health;mechanisms of carcinogenesis;Mice;Mice,Inbred C3H;Mutagenesis;Oxidative Stress;pharmacology;Safety;Tetradecanoylphorbol Acetate.
ISSN:
0027-5107
Appears in Collections:
Articles with annotation

Full metadata record

DC FieldValue Language
dc.contributor.authorParfett, Craig L.-
dc.contributor.authorHealy, Caroline-
dc.date.accessioned2009-05-18T07:10:44Z-
dc.date.available2009-05-18T07:10:44Z-
dc.date.issued2006-04-30-
dc.identifier.citationMutat. Res. 2006, 604 (1-2):42-52en
dc.identifier.issn0027-5107-
dc.identifier.pmid16459133-
dc.identifier.doi10.1016/j.mrgentox.2005.12.008-
dc.identifier.urihttp://hdl.handle.net/10146/68416-
dc.descriptionKEYWORDS - CLASSIFICATION: Animals;Base Sequence;chemistry;Canada;Carcinogens;Cell Culture Techniques;Cell Survival;Cells,Cultured;Chromosomal Instability;Clone Cells;drug effects;Dna;DNA Fingerprinting;DNA Primers;Environmental Health;mechanisms of carcinogenesis;Mice;Mice,Inbred C3H;Mutagenesis;Oxidative Stress;pharmacology;Safety;Tetradecanoylphorbol Acetate.en
dc.description.abstractThe ability of tumour promoters to alter DNA stability within regions that contain tandemly repeated sequences (TRSs), was studied in a cell culture model of multi-stage carcinogenesis. Non-cytotoxic concentrations of TPA (12-O-tetradecanoyl-phorbol-13-acetate) and xanthine oxidase with xanthine substrate, sufficient to promote morphological transformation in C3H/10T1/2 cultures, were tested for their effects on mutation frequencies in TRSs by a DNA fingerprinting approach. Specifically, restriction digests of genomic DNA samples from randomly selected, non-transformed clones, isolated from cultures after several days exposure to promoters, were visualized by Southern hybridizations with the multi-locus pentamer repeat sequence probe, Ms6-Hm (Pc-1). Basal and promoter-induced frequencies of sub-clone TRS fingerprint polymorphisms were estimated in five cell populations: an uncloned stock culture, three populations established from normal-appearing sub-clones, and one clonal population established from a 3-methylcholanthrene (MCA)-transformed focus. Basal variant fingerprint frequencies spanned a range from 0.0 to 0.43% mutants/band among cells from the four untransformed populations. Both TPA and xanthine oxidase treatments significantly increased recorded mutation frequencies, 2.3- and 2.7-fold, respectively, using combined data from the progenitor population and three untransformed clones. The untreated MCA-transformed clonal population appeared to contain a single, pre-existing mutant restriction fragment, but additional mutations were induced thereafter, in response to the promoting treatments. Taken together, the measured increases in mutations were highly significant and suggest that promoters of cell transformation in the C3H/10T1/2 cell line might induce a genome-wide instability targeted to regions containing Ms6-Hm sequence motifs.en
dc.language.isoenen
dc.relation.urlhttp://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T2D-4J625TV-1&_user=1843694&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000055040&_version=1&_urlVersion=0&_userid=1843694&md5=537d40f05453dcca95ca0161f896e2d1en
dc.subjectDNA fingerprintingen
dc.subjectMulti-locus probesen
dc.subjectNon-mutagenicen
dc.subjectOxidative stressen
dc.subjectTumour promotionen
dc.subject.meshAnimals-
dc.subject.meshBase Sequence-
dc.subject.meshCarcinogens-
dc.subject.meshCell Culture Techniques-
dc.subject.meshCell Survival-
dc.subject.meshCells, Cultured-
dc.subject.meshChromosomal Instability-
dc.subject.meshClone Cells-
dc.subject.meshDNA-
dc.subject.meshDNA Fingerprinting-
dc.subject.meshDNA Primers-
dc.subject.meshMice-
dc.subject.meshMice, Inbred C3H-
dc.subject.meshOxidative Stress-
dc.subject.meshTetradecanoylphorbol Acetate-
dc.titleTandemly repeated DNA sequence instabilities induced by two promoters of morphological transformation in vitro: a short-term response to non-mutagenic agents in C3H/10T1/2 cells.en
dc.typeArticleen
dc.identifier.journalMutation researchen
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