Comparison of p53 protein expression and cellular localization in human and hamster pancreatic cancer cell lines.
We compared the expression of p53 protein in four human pancreatic cancer cell lines (HPAF, CD11, CD18 and PANC-1) and four hamster pancreatic cancer cell lines (PC-1, PC-1.2, PC-1.0 and H2T) by the monoclonal antibodies PAb421 and PAb240. PAb421 reacted with all human pancreatic cancer cell lines but not with the hamster cells. PAb240, on the other hand, reacted with all human and hamster pancreatic cancer cell lines in immunoblotting and in immunocytochemistry. However, immunoprecipitation with PAb240 was detected only in human cell lines HPAF, CD11 and CD18 cells but not in PANC-1 or in any of the hamster cell lines. During exponential growth, immunoreactivity was detected mainly in the nucleus of PC-1, PC-1.2 and PANC-1 cells (nuclear type) and in both the nucleus and the cytoplasm of PC-1.0, H2T, HPAF, CD11 and CD18 cells (diffuse type). At the confluence, the expression of p53 was decreased in most of the human cell lines as was proliferative cell nuclear antigen. After incubation with 1 mM hydroxyurea, cells with nuclear p53 expression did not show an altered cellular distribution of p53 protein, whereas cells with a diffuse type of localization pattern showed an increase in the nuclear staining. On the other hand, cytoplasmic immunoreactivity was found in PC-1.0, PC-1, PC-1.2, HPAF, CD11 and CD18 cells that were treated with 100 ng/ml of nocodazole. After heat stress with 1 h incubation at 42 degrees C, p53 protein was detected in the cytoplasm and nucleolus of all cell lines. After 24-48 h incubation at 37 degrees C, this change in cellular distribution of p53 in response to heat stress was reverted to a preheat stress pattern. The overall results suggest that neither the p53 of PANC-1 nor the hamster pancreatic cancer cell lines are immunoprecipitated with the PAb240. It appears that cell cycle and heat stress are two of the factors that influence cellular localization of p53 protein in both human and hamster pancreatic cancer cells.
Mogaki
M
M
Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha 68198-6805.
Hirota
M
M
Chaney
W G
WG
Pour
P M
PM
D000818:Animals; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D006358:Hot Temperature; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D007150:Immunohistochemistry; D010190:Pancreatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D011233:Precipitin Tests; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis*
D016159:Tumor Suppressor Protein p53; D006918:Hydroxyurea