A Method for Detecting Carcinogenic Organic Chemicals Using Mammalian Cells in Culture

Overview

Journal Br J Cancer

Specialty Oncology

Date 1977 Nov 1

PMID 588420

Citations 18

Authors

J A Styles

Affiliations

Soon will be listed here.

Abstract

A method for testing organic chemicals for their carcinogenic potential is described. Baby hamster kidney cells (BHK-21/C1 13) were exposed to different doses of test compound in liquid tissue culture medium containing rat liver post-mitochondrial supernatant and cofactors (S-9 mix) to aid metabolism, but without serum. Survival of cells following exposure to the compound was assessed by cloning in liquid growth medium. Transformation was assessed by colony growth in semi-solid agar. The dose-response curve for survival was used to determine the LC50 of the compound. A dose-response curve for transformation was constructed and a 5-fold increase in transformation frequency at the LC50 was regarded as a positive test result. The method may also be used for testing gaseous compounds. Cells grown in monolayers and overlaid with serum-free medium and S-9 mix were exposed to vinyl chloride gas mixed with air. After exposure, the treated cells were trypsinized, resuspended in growth medium, and survival and transformation assays performed. The methods described are illustrated by examples taken from an evaluation study using 120 compounds and found to be more than 90% accurate in distinguishing between carcinogens and non-carcinogens.

Citing Articles

Immortalization of chicken preadipocytes by retroviral transduction of chicken TERT and TR.

Wang W, Zhang T, Wu C, Wang S, Wang Y, Li H PLoS One. 2017; 12(5):e0177348.

PMID: 28486516 PMC: 5423695. DOI: 10.1371/journal.pone.0177348.

In vitro assessment of the toxicity of metal compounds : I. Mammalian Cell transformation.

Heck J, Costa M Biol Trace Elem Res. 2013; 4(2-3):71-82.

PMID: 24271980 DOI: 10.1007/BF02783248.

Premalignant and neoplastic skin lesions associated with occupational exposure to "tarry" byproducts during manufacture of 4,4'-bipyridyl.

Bowra G, DUFFIELD D, Osborn A, Purchase I Br J Ind Med. 1982; 39(1):76-81.

PMID: 7066224 PMC: 1008931. DOI: 10.1136/oem.39.1.76.

Mutagenic and cell transforming activities of 1-chlor-2,4-dinitrobenzene (DNCB) and squaric-acid-dibutylester (SADBE).

Strobel R, ROHRBORN G Arch Toxicol. 1980; 45(4):307-14.

PMID: 7004402 DOI: 10.1007/BF00293811.

Procedures for screening chemicals for carcinogenicity.

Purchase I Br J Ind Med. 1980; 37(1):1-10.

PMID: 6989387 PMC: 1008639. DOI: 10.1136/oem.37.1.1.

References

Macpherson I, Montagnier L . AGAR SUSPENSION CULTURE FOR THE SELECTIVE ASSAY OF CELLS TRANSFORMED BY POLYOMA VIRUS. Virology. 1964; 23:291-4. DOI: 10.1016/0042-6822(64)90301-0. View

Purchase I, Longstaff E, Ashby J, Styles J, Anderson D, Lefevre P . Evaluation of six short term tests for detecting organic chemical carcinogens and recommendations for their use. Nature. 1976; 264(5587):624-7. DOI: 10.1038/264624a0. View

BOUCK N, DI MAYORCA G . Somatic mutation as the basis for malignant transformation of BHK cells by chemical carcinogens. Nature. 1976; 264(5588):722-7. DOI: 10.1038/264722a0. View

DI MAYORCA G, Greenblatt M, Trauthen T, Soller A, Giordano R . Malignant transformation of BHK21 clone 13 cells in vitro by nitrosamines--a conditional state. Proc Natl Acad Sci U S A. 1973; 70(1):46-9. PMC: 433180. DOI: 10.1073/pnas.70.1.46. View

Bartsch H, Malaveille C, Montesano R . Human, rat and mouse liver-mediated mutagenicity of vinyl chloride in S. typhimurium strains. Int J Cancer. 1975; 15(3):429-37. DOI: 10.1002/ijc.2910150309. View

Mishra N, DI MAYORCA G . In vitro malignant transformation of cells by chemical carcinogens. Biochim Biophys Acta. 1974; 355(3-4):205-19. DOI: 10.1016/0304-419x(74)90010-9. View

MALTONI C, Lefemine G, Chieco P, CARRETTI D . Vinyl chloride carcinogenesis: current results and perspectives. Med Lav. 1974; 65(11-12):421-44. View

Wogan G, Paglialunga S, ARCHER M, Tannenbaum S . Carcinogenicity of nitrosation products of ephedrine, sarcosine, folic acid, and creatinine. Cancer Res. 1975; 35(8):1981-4. View

Creech Jr J, Johnson M . Angiosarcoma of liver in the manufacture of polyvinyl chloride. J Occup Med. 1974; 16(3):150-1. View

10.

Kirkland D, Pick C . The histological appearance of tumours derived from rat embryo cells transformed in vitro spontaneously and after treatment with nitrosomethylurea. Br J Cancer. 1973; 28(5):440-52. PMC: 2008912. DOI: 10.1038/bjc.1973.170. View

11.

Umeda M, Iype P . An improved expression of in vitro transformation rate based on cytotoxicity produced by chemical carcinogens. Br J Cancer. 1973; 28(1):71-4. PMC: 2009073. DOI: 10.1038/bjc.1973.72. View

12.

Ames B, McCann J, Yamasaki E . Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. Mutat Res. 1975; 31(6):347-64. DOI: 10.1016/0165-1161(75)90046-1. View

13.

Kirkland D, Armstrong C, Harris R . Spontaneous and chemically induced transformation of rat embryo cell cultures. Br J Cancer. 1975; 31(3):329-37. PMC: 2009414. DOI: 10.1038/bjc.1975.67. View

14.

Evans C, DIPAOLO J . Neoplastic transformation of guinea pig fetal cells in culture induced by chemical carcinogens. Cancer Res. 1975; 35(4):1035-44. View

15.

VIOLA P, Bigotti A, Caputo A . Oncogenic response of rat skin, lungs, and bones to vinyl chloride. Cancer Res. 1971; 31(5):516-22. View