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To investigate how a carcinogenic promoter acts on cells mutated by an initiator, we used as a model, lacZ transgenic mouse and a positive selection system. Preliminary data for the mutational events in liver DNA of the mice was generated using diethylnitrosamine (DEN) and sodium phenobarbital (S-PB) as initiator and promoter, respectively. In our first experiment, male MutaMice received a single i.p. injection of saline or 100 mg/kg DEN and were fed a normal diet for 7 days and 500 p.p.m. S-PB in the diet for 21 days. Liver DNA was harvested after a 1 night fast on days 7 and 28 post-DEN treatment. In our second experiment, male mice received a single i.p. injection of phosphate buffered saline or 50 mg/kg DEN and were fed a normal diet for 7 days, a diet with S-PB for 14 days and then a normal diet for 7 days. Liver DNA was harvested after a 1 night fast on days 7, 21 and 28 post-DEN treatment. The S-PB diet enhanced absolute and relative liver weights in all groups. The single intraperitoneal dose of 50 or 100 mg/kg DEN induced high mutation frequencies (MF) in liver, lacZ genes on days 7, 21 and 28. There were no remarkable differences of the MF among any sampling days for animals receiving DEN and a normal diet. S-PB feeding at 500 p.p.m. for 21 days failed to affect the MF in groups given saline or 100 mg/kg DEN. On the other hand, when 50 mg/kg DEN was given, S-PB feeding at 500 p.p.m. for 14 days elevated the MF in liver DNA on days 21 and 28 to approximately 1.8 and 4.0 times the MF, respectively, of the mice fed the normal diet. Consequently, S-PB might preferentially promote certain initiated cells participating in a balance between cell death and proliferation.

Sodium phenobarbital-enhanced mutation frequency in the liver DNM of lacz transgenic mice treated with diethylnitrosamine