Reduced expression of mature TGF beta 1 correlates with the suppression of rat hepatocyte apoptosis by the peroxisome proliferator, nafenopin.
Non-genotoxic carcinogens cause cancer without damaging the DNA. Peroxisome proliferators (PPs) are a class of potent rodent non-genotoxic hepatocarcinogens that may act by perturbing hepatocyte growth regulation. Previously, we have shown that although cultured rat hepatocytes degenerate rapidly in culture, their survival can be reversibly maintained by the PP nafenopin. This prolonged survival is associated with a decrease in the number of hepatocytes displaying the chromatin condensation characteristic of apoptosis. The addition of the negative growth regulator TGF beta-1 induced high levels of hepatocyte apoptosis but nafenopin was able to suppress this TGF beta-1 induced apoptosis. These data suggested that increased levels of mature TGF beta-1 may be involved in the signalling of the apoptosis seen in degenerating hepatocyte cultures. To test this hypothesis, we carried out Western blot analyses using a anti-TGF beta 1 antibody. There was an increase (p = 0.014) in expression of mature TGF beta 1 in degenerating rat hepatocyte cultures compared with hepatocyte cultures surviving in the presence of nafenopin. However, there was a concomitant decrease (p = 0.024) in TGF beta 1-latency activated protein (TGF beta 1-LAP), the precursor of the active, mature form. Immunocytochemistry confirmed that TGF beta 1/TGF beta 1-LAP expression was predominantly in the hepatocytes displaying apoptotic morphology although expression was detected also in non-parenchymal liver cells. The immunocytochemistry data indicate that TGF beta 1 is involved during the onset of hepatocyte apoptosis and that the PP nafenopin can impinge on this cell death pathway. TGF beta 1-LAP, probably produced mainly by the non-parenchymal liver cells, may be processed less efficiently to the mature, active form in the presence of nafenopin, although more data are required to confirm this hypothesis.
Strange
J
J
Zeneca Central Toxicology Laboratory, Alderley Park, Macclesfield, UK.
Roberts
R A
RA
D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D015153:Blotting, Western; D002273:Carcinogens / Q000494:pharmacology; D002478:Cells, Cultured; D002843:Chromatin / Q000187:drug effects; D000960:Hypolipidemic Agents / Q000494:pharmacology; D007150:Immunohistochemistry; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D008830:Microbodies / Q000187:drug effects; D009255:Nafenopin / Q000494:pharmacology*; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D013194:Staining and Labeling; D016212:Transforming Growth Factor beta / Q000096:biosynthesis* / Q000187:drug effects / Q000276:immunology
D002273:Carcinogens; D002843:Chromatin; D000960:Hypolipidemic Agents; D016212:Transforming Growth Factor beta; D009255:Nafenopin