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KMID : 0377519950200030215
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Chung-Ang Journal of Medicine
1995 Volume.20 No. 3 p.215 ~ p.235
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Analysis of Glutathione S-transferase,AgNORs and DNA ploidy during the Hepatocarcinogenesis induced by N,N-diethylnitrosamine in Rat
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Park Eon-Sub
Kim Mi-Kyung
Yoo Jae-Hyung
Song Kye-Yong
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Abstract
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To investigate the transforming potential in various hyperplastic nodules developed during hepatocarcinogenesis induced by N, N-diethylnitrosamine(DEN), we analyzed the sequential histopathology, foci of localization of Glutathione S-Transferase(GST-P), numbers of AgNORs and DNA ploidy in various nodules as well as in carcinoma. Male rats, Wister, with body weight of 150-170gm were used and fed daily oral intak by 150ppm DEN. Five animals were killed at an interval of two weeks during experimental 16 weeks period and at the same date two control rats were killed for comparison. Areas of Gluathione S-transferase foci were analyzed by image analysis, numbers of AgNORs were analyzed by direct counting under the light microscope and DNA ploidy by flow cytometry. Immunohistochemical staining was done using avidin biotin complex with nonspecific polyclonal GST-P antibody. The clear cell nodule appeared first at the 3rd weeks after DEN administration followed by eosinophilic and basophilic nodules whit mixed cell nodules consisting respective cells in between, and carcinoma appears at the 14 weeks. GST immunostaining were revealed a tendency of progressive increase in positive foci, strong reaction were noted especially in clear cells and eosinophilic cell nodules as well as in addenoma and carcinoma. Mean numbers of AgNORs per nucleus in control, clear cell, eosinophilic cell, basophilic cell nodule, addenoma and carcinoma were 1.59, 2.86, 2.66, 2.72, 2.92 and 3.43, respectively. And the perentage of cells containing over five AgNORs were 0.07%, 7.30%, 6.20%, 6.50%, 8.80% and 12.40%(p<0.01), respectively. The percentage of aneuploidy were 14.3%, 36.4%, 30.0%, 40.0% and 45.5%, in respective nodules. As above results it could be suggested that carcinoma night be developed from the eosinophilic cell nodule through the aberration of DNA ploidy as in basophilic cell nodule and adenoma in the hepatocarcinogensis.
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KEYWORD
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carcinogenesis, N, N-Diethylnitrosamine, DNA ploidy, AgNORs, glutation S-transferase, hepatocellular carcinoma
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