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(Received for publication, June 30, 1995; and in revised form, November 27, 1995)
Transforming growth factor-
(TGF-), a growth regulator
of fetal hepatocytes in primary culture, also regulates death of these
cells. Dose-response analysis showed that the TGF- concentration
needed to induce hepatocyte death (2.5 ng/ml) was 5 times that needed
to inhibit growth in these cells (0.5 ng/ml). In response to TGF-,
hepatocytes induced DNA fragmentation and the appearance of nuclei with
a DNA content lower than 2C (diploid content), typical of a programmed
cell death model. TGF--induced apoptosis in fetal hepatocytes was
preceded by an induction of reactive oxygen species production and a
decrease in the glutathione intracellular content, indicating that this
factor induces oxidative stress in fetal hepatocytes. Studies performed
to analyze levels of c-fos mRNA, a gene whose expression is
modulated by redox state, demonstrated that only high, apoptotic
concentrations of TGF- (2.5 ng/ml) produced an increase in the
mRNA levels of this gene, the level of induction being similar to that
found when cells were incubated in the presence of tert-butyl
hydroperoxide. Gel mobility shift assays showed that the
c-fos-induced expression was coincident with an increase in
AP-1 activity. Finally, cell death induced by TGF- in fetal
hepatocytes was partially blocked by radical scavengers, which
decreased the percentage of apoptotic cells, whereas these agents did
not modify the growth-inhibitory effect elicited by TGF- in these
cells. In summary, the results presented in this paper provide evidence
for the involvement of an oxidative process in the apoptosis elicited
by TGF- in fetal hepatocytes.
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