Epithelial Focus Assay for Early Detection of Carcinogen-Altered Cells in Various Organs of Rats Exposed In Situ to N-Nitrosoheptamethyleneimine

Abstract

The purpose of these studies was to determine a) whether epithelial cells with altered in vitro growth capacity occur not only after topical application of 7–12-dimethylbenz[a]-anthracene but also after systemic administration of a carcinogenic nitrosamine, and b) whether such cells can be isolated from tissues other than tracheal mucosa. At 3 and 20 weeks following intragastric administration of 150, 300, or 600 mg N-nitrosohepatamethyleneimine (NHMI)/kg, cells were harvested from tracheas, esophagi, and lungs (target tissues for NHMI) of inbred F344 rats and seeded into culture dishes. Normal cells from nonexposed organs produced no proliferative epithelial foci (EF). Of those tracheas sampled 3 weeks following exposure to 150 and 300 mg/kg, 10 and 20%, respectively, contained one or more EF that could be subcultured. Of those tracheas harvested 3 weeks post exposure to 600 mg/kg or 20 weeks post exposure to 150–600 mg/kg, 80–100% contained EF that could be subcultured. Twenty weeks after 600 mg NHMI/kg, the incidence of tracheas harboring cell populations with neoplastic potential (agarose-positive EF) was 80%, whereas the tracheal tumor incidence determined at 24 months was only 29%. Epithelial focus-forming units with various abnormal in vitro growth potentials were also detected in esophagi and lungs of NHMI-exposed rats.