Exogenous epidermal growth factor exerts promoting action during the early phase of rat urinary bladder carcinogenesis

Using the heterotopically transplanted rat urinary bladder (HTB) model that was developed in our laboratory, we examined the relationship between the duration of epidermal growth factor (EGF) treatment and acquisition of EGF-independence of urinary bladder tumors that were induced by EGF stimulation. After treatment with N-methyl-N-nitrosourea (MNU) (0.25 mg/0.5 ml of 0.9% NaCl once a week for 3 consecutive weeks), animals at week 3 received EGF [250 ng/0.5 ml phosphate-buffered saline (PBS)] into the HTBs once a week for 20, 28, or 36 weeks. For examination of the effect of EGF withdrawal, one half of the rats received the vehicle (PBS) only beginning at week 23 or week 31 for 8 weeks. When animals were examined at week 23, the incidence and the mean number of tumors per bladder were low, irrespective of EGF treatment. In the bladders that had been exposed to EGF during the first 20 weeks after MNU administration, however, both the incidence and the mean number of tumors per bladder had increased significantly at week 31, regardless of whether or not EGF treatment was continued beyond week 23. Between weeks 31 and 39, EGF treatment demonstrated no effect; both the incidence of tumors and the mean number of tumors were the same as those at week 31. These results suggest that EGF exerts its promoting effect only during the early phase of MNU-initiated bladder carcinogenesis, but that its effect becomes manifest during the subsequent 8 weeks. EGF independence may be due to establishment of an autocrine growth-stimulatory mechanism in bladder tumors.