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© 1991 Oxford University Press

other

In vitro tramsformation of rat bladder epithelium by 2-amino-4-(5-nitro-2-furyl)thiazole

Angela M. Mann 1, Tsuneo Masui 1, Frank J. Chlapowski 3, Takehiko Okamura 1, Claudia D. Borgeson 1 and Samuel M. Cohen 1 2 4

1Department of Pathology and Microbiology
2Eppley Institute, University of Nebraska Medical Center Omaha, NE
3Department of Biochemistry, University of Massachusetts Medical Center Worcester, MA, USA

4To whom reprint requests should be sent at: Department of Pathology and Microbiology, University of Nebraska Medical Center, 600 South 42nd Street, Omaha, NE 68198-3135. USA

To establish a rat urinary bladder carcinogenesis model in vitro, primary rat bladder epithelial cells were grown in media containing 2-amino-4(5-nitro-2-furyl)thiazole (ANFT), the water-soluble metabolite of N-[4-(5-nitro-2-furyl)-2-thiazolyl]-formamide (FANFT), for 4 weeks followed by longterm (4-7 months) exposure to control medium, sodium saccharin (NaS), or urea. Another set of cultures were exposed to ANFT, NaS and urea simultaneously. Several phenotypic changes were observed in the chemically exposed cell cultures, namely differences in cell morphology, increased growth rate and the ability to grow on plastic instead of rattail collagen support. All of the chemically exposed cultures were anchorage independent except one of those treated with NaS. The ANFT-treated cells followed by control medium or urea and cells treated with ANFT, NaS and urea were tumorigenic when transplanted to nude mice, whereas NaS or ANFT followed by NaS treatment were not. The tumors were carcinomas and their epithelial differentiation was verified by strong positive staining for cytokeratin. These studies demonstrate the urothelial transforming capability of ANFT in cell culture without the necessity for a long exposure to a secondary chemical.


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