This study was designed to evaluate the effect of cyclophosphamide(CY) on diethylnitrosamine(DEN)-induced hepatic tumors in rats.
Thirty five male or female Sprague Dawley rats were continiously given water containing 0.01% DEN for 10 weeks and then were given with CY 25㎎/rat/day in water for 3, 5, 7 or 9 days.
The livers of rats were removed and fixed in 10% buffered neutral formalin.
The appearances of positive cells by immunohistochemical methods using proliferating cell nuclear antigen (PCNA) antibody, p53 antibody and apoptotic kit were investigated.
The livers of rats given with CY were grossly brilliant, red-brown color, flexible, and thin border, and stainability of the liver cells were restored microscopically, and the vaccuolated and degenerated regions were differentiated from restored regions. These restored findings also were advanced in control group because of no DEN treatment but tended to be less avanced.
In immunohistochemistry, positive cells to PCNA antibody appeared more numerous in control groups than that of CY treated groups. Appearance of positive cells in CY-treated group for 7 days and for 9 days were more numerous than those of CY-treated groups for 3 days and for 5 days, respectively. So these findings suggested that CY suppressed cell proliferations and effects of these action were decreased with CY-treated days.
The numbers of positive cells to PCNA antibody were more prominent in hepatocellular carcinoma regions and cholangiocarcinoma regions, and then were ranked as order of large liver cell regions and normal liver cell regions.
Also the numbers of the positivc cells by apoptotic kit tended to be higher in hepatocellular carcinoma regions and cholangiocarcinoma regions but not uniformly in order in all regions and were much less numbers than those of PCNA positive cells. So immunohistochemical methods using PCNA antibody together than using apoptotic kit alone when anti-carcinogen experiments.
Rats with positive cells by p53 antibody were 11 of 15 rats(73.4%) in control groups and 12 of 18 rats(66.7%) in CY treated group, respectively. These positive cells appeared focally in early vacuole-occurring regions and were very low in numbers.
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