Ras involvement in cells transformed with 2-amino-4-(5-nitro-2-furyl)thiazole (ANFT) in vitro and with N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide in vivo

Angela M. Mann, Makoto Asamoto, Frank J. Chlapowski, Tsuneo Masui, Timothy L. Macatee, Samuel Monroe Cohen

Research output: Contribution to journalArticle

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Abstract

N-[4-(5-nitro-2-furyl)-2-thiazolyl]fonnamide (FANFT) administration to rats followed by sodium saccharin results in transitional cell carcinomas of the bladder, of which 24% harbor an activated H-ras gene. Since 2-amino-4-(5-nitro-2-furyI)thiazole (ANFT) is the mutagenic and carcinogenic metabolite of FANFT in vivo, we wished to examine ras activation in in vitro ANFT-transformed rat bladder epithelial cells as well as four cell lines established in culture from in vivo FANFT-induced rat bladder tumors. Screening by Western blotting revealed no enhanced levels of p21ras in ANFT-transformed cells nor in cells established hi culture from FANFT-induced rat bladder carcinomas. Further investigations using immunohistochemical staining with a different pan-reactive p21 monoclonal antibody (Cetus Corporation) specific for this method, however, showed two groups of cells from FANFT-induced rat bladder tumors had enhanced immunoreactivity. Apart from this, p21ras expression of most of the cell groups varied little from the controls. We examined the reported hot spots (exons 1 and 2) of each of the ras genes (H-, K- and N-ras) by direct sequencing of amplified DNA. No mutations were present. We conclude, therefore, that ANFT transformation of primary rat bladder epithelial cells in vitro may not in this case be mediated by ras activation, although this is difficult to determine since others have observed that optimal culture conditions can select for certain populations of cells without ras activation.

Original languageEnglish (US)
Pages (from-to)1651-1655
Number of pages5
JournalCarcinogenesis
Volume13
Issue number9
DOIs
StatePublished - Sep 1 1992

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FANFT
ras Genes
Urinary Bladder
Urinary Bladder Neoplasms
Epithelial Cells
Thiazoles
Saccharin
Transitional Cell Carcinoma
DNA Sequence Analysis
formamide
In Vitro Techniques
ANFT
Exons
Western Blotting
Monoclonal Antibodies
Staining and Labeling
Carcinoma
Cell Line
Mutation
Population

ASJC Scopus subject areas

  • Cancer Research

Cite this

Ras involvement in cells transformed with 2-amino-4-(5-nitro-2-furyl)thiazole (ANFT) in vitro and with N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide in vivo. / Mann, Angela M.; Asamoto, Makoto; Chlapowski, Frank J.; Masui, Tsuneo; Macatee, Timothy L.; Cohen, Samuel Monroe.

In: Carcinogenesis, Vol. 13, No. 9, 01.09.1992, p. 1651-1655.

Research output: Contribution to journalArticle

Mann, Angela M. ; Asamoto, Makoto ; Chlapowski, Frank J. ; Masui, Tsuneo ; Macatee, Timothy L. ; Cohen, Samuel Monroe. / Ras involvement in cells transformed with 2-amino-4-(5-nitro-2-furyl)thiazole (ANFT) in vitro and with N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide in vivo. In: Carcinogenesis. 1992 ; Vol. 13, No. 9. pp. 1651-1655.
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abstract = "N-[4-(5-nitro-2-furyl)-2-thiazolyl]fonnamide (FANFT) administration to rats followed by sodium saccharin results in transitional cell carcinomas of the bladder, of which 24{\%} harbor an activated H-ras gene. Since 2-amino-4-(5-nitro-2-furyI)thiazole (ANFT) is the mutagenic and carcinogenic metabolite of FANFT in vivo, we wished to examine ras activation in in vitro ANFT-transformed rat bladder epithelial cells as well as four cell lines established in culture from in vivo FANFT-induced rat bladder tumors. Screening by Western blotting revealed no enhanced levels of p21ras in ANFT-transformed cells nor in cells established hi culture from FANFT-induced rat bladder carcinomas. Further investigations using immunohistochemical staining with a different pan-reactive p21 monoclonal antibody (Cetus Corporation) specific for this method, however, showed two groups of cells from FANFT-induced rat bladder tumors had enhanced immunoreactivity. Apart from this, p21ras expression of most of the cell groups varied little from the controls. We examined the reported hot spots (exons 1 and 2) of each of the ras genes (H-, K- and N-ras) by direct sequencing of amplified DNA. No mutations were present. We conclude, therefore, that ANFT transformation of primary rat bladder epithelial cells in vitro may not in this case be mediated by ras activation, although this is difficult to determine since others have observed that optimal culture conditions can select for certain populations of cells without ras activation.",
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AU - Mann, Angela M.

AU - Asamoto, Makoto

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