Effect of three tumour promoters on the stability of hepatocyte cultures and apoptosis after transforming growth factor-beta 1.
Tumour promoters like the anti-androgen cyproterone acetate (CPA), the peroxisome proliferator nafenopin (NAF) and phenobarbital (PB) stimulate liver growth in rodents. Transforming growth factor-beta 1 (TGF-beta 1) is expressed in livers after treatment with CPA (Oberhammer et al., submitted) and some peroxisome proliferators. In this paper we describe the influence of CPA, NAF and PB on the stability of hepatocyte cultures and induction of apoptosis by TGF-beta 1. All three tumour promoters had a stabilizing effect on confluent monolayers of hepatocytes, partially preventing the usually occurring dedifferentiation and detachment processes. CPA on its own was able to induce apoptosis at the high dose of 10 microM. No induction of apoptosis could be observed after PB and NAF. At any dose above 0.01 microM CPA enhanced TGF-beta 1-induced apoptosis (5.8-fold increase with 10 microM CPA). Thus the combination of 10 microM CPA and 1 ng/ml TGF-beta 1 induced apoptosis in 90% of the plated hepatocytes. At a high dose (10 microM) NAF produced a 35% reduction in apoptosis induced by TGF-beta 1, in parallel with a stabilizing effect on cell number. PB did not affect the rate of apoptosis induced by TGF-beta 1. As demonstrated by immunohistochemical detection of PCNA, TGF-beta 1 prevented induction of PCNA by epidermal growth factor (EGF). No induction of PCNA was observable in CPA-treated cultures. In untreated and EGF-treated cultures TGF-beta 1 was able to induce apoptosis to the same extent within 30 h. In CPA-treated cultures this period was shortened to 12 h. Thus CPA shortens the lag phase of induction of apoptosis by shifting hepatocytes to a point before S phase, where they are highly susceptible to TGF-beta 1-induced apoptosis.
Oberhammer
F A
FA
Institute of Tumorbiology, Vienna, Austria.
Qin
H M
HM
D017209:Apoptosis / Q000187:drug effects*; D002273:Carcinogens / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D017373:Cyproterone Acetate / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000494:pharmacology; D066298:In Vitro Techniques; D008099:Liver / Q000166:cytology / Q000187:drug effects*; D009255:Nafenopin / Q000494:pharmacology; D010634:Phenobarbital / Q000494:pharmacology; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D013997:Time Factors; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D002273:Carcinogens; D018809:Proliferating Cell Nuclear Antigen; D016212:Transforming Growth Factor beta; D009255:Nafenopin; D017373:Cyproterone Acetate; D004815:Epidermal Growth Factor; D010634:Phenobarbital