Immunohistochemical visualization of wild-type p53 protein in paraffin- embedded rat liver using tyramide amplification: zonal hepatic distribution of p53 protein after N-hydroxy-2-acetylaminofluorene administration |
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Authors: | van Gijssel HE; van Gijlswijk RP; de Haas RR; Stark C; Mulder GJ; Meerman JH |
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Institution: | Division of Toxicology, Leiden Amsterdam Center for Drug Research, The Netherlands. gijssel@lacdr.leidenuniv.nl |
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Abstract: | P53 protein plays an important role in regulation of the cell cycle.
Recently, a role in tumour genesis has also been suggested. The protein is
induced after various forms of DNA damage. Immunohistochemical detection of
p53 protein showed positive cells in human skin after UV- irradiation, in
mouse skin after benzoa]pyrene treatment and in mouse spleen, thymus and
bone after gamma-irradiation. However, no staining was found in mouse and
rat liver with traditional immunohistochemical staining methods due to the
low amount of p53 present. This seriously hampered studies on the role of
p53 in hepatocarcinogenesis. We have developed a more sensitive
immunohistochemical method for staining of p53 in paraffin-embedded
sections of rat liver using microwave irradiation for antigen retrieval,
avidin-biotin complexing and tyramide amplification. A strong, specific
fluorescence signal for p53 was found in hepatocytes of rats that had
received the hepatocarcinogen N-hydroxy-2-acetylaminofluorene; in control
liver no such p53 staining was observed. The fluorescence was located in
the nucleus of hepatocytes in zone 1 of the liver. This agrees with the
fact that N- hydroxy-2-acetylaminofluorene causes cytotoxicity in this
zone.
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