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Crocidolite induces cell transformation and p53 gene mutation in BALB/c-3T3 cells.

Authors
Lin-F; Liu-YG; Liu-YQ; Keshava-N; Li-S
Source
Teratog, Carcinog, Mutagen 2000 Sep-Oct; 20(5):273-281
NIOSHTIC No.
20020735
Abstract
Cell transformation is one of the most common assays used to study morphological changes in the multistep process of carcinogenesis. The present study was initiated to investigate the ability of crocidolite to induce cell transformation in BALB/c-3T3 cells and to analyze the relationship between p53 mutations and crocidolite-induced cell transformation, if any. Cell transformation was carried out according to standard procedures. Exponentially growing cells were exposed to different concentrations (0.2-20 microg/cm(2)) of crocidolite fibers for 72 h. Foci obtained from cell transformation were analyzed for their ability to grow in soft agar (anchorage-independence) and p53 alterations. The results of this study demonstrate that there was an increase in transformation frequency (TF) with an increase in concentration of crocidolite. Also, focal cells were able to grow on soft agar, indicating anchorage-independence. cDNA was prepared from RNA isolated from Type 3 foci and subjected to mutational analysis. Eleven exons of the p53 gene from eight transformed cell lines were analyzed for alterations using polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP). Alterations were found in seven of eight cell lines, two of them were in exons 4-6, and five in exons 9-11. The alterations were randomly scattered among the crocidolite dose groups. These results suggest that crocidolite induces mutations predominantly in exons 9-11 of the p53 gene in a nondose-dependent manner.
Keywords
Cell-transformation; Gene-mutation; Carcinogenesis; Carcinogenicity; Carcinogens; Morphology; Exposure-assessment
Contact
Nagalakshmi Keshava, Ph.D., Toxicology and Molecular Biology Branch, HELD, NIOSH, M/S 3014, 1095 Willowdale Road, Morgantown, WV 26505
CODEN
TCMUD8
Publication Date
20000901
Document Type
Journal Article
Email Address
ndk2@cdc.gov
Fiscal Year
2000
NTIS Accession No.
NTIS Price
Issue of Publication
5
ISSN
0270-3211
NIOSH Division
HELD
Source Name
Teratogenesis, Carcinogenesis, and Mutagenesis
State
WV
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