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* Departamento de Bioquímica y Biología Molecular, Instituto de Bioquímica, Centro Mixto CSIC/UCM and
Centro de Citometría de Flujo y Microscopía Confocal, Facultad de Farmacia, Universidad Complutense de Madrid, 28040 Madrid, Spain
1Correspondence: Dpto. de Bioquímica y Biología Molecular, Facultad de Farmacia, Universidad Complutense de Madrid, 28040 Madrid, Spain. E-mail: isabelf{at}eucmax.sim.ucm.es
Treatment of fetal rat hepatocytes with transforming growth factor beta
(TGF-ß) is followed by apoptotic cell death. Analysis of radical
oxygen species (ROS) content and mitochondrial transmembrane potential
(
m), using specific fluorescent probes in FACScan and
confocal microscopy, showed that TGF-ß mediates ROS production that
precedes the loss of m, the release of cytochrome
c, and the activation of caspase 3. TGF-ß induces a
decrease in the protein and mRNA levels of bcl-xL, an
antiapoptotic member of the Bcl-2 family. In contrast, there is no
change in the expression and/or translocation of Bax, a proapoptotic
member of the same family. EGF maintains Bcl-xL, preventing
m collapse and release of cytochrome
c. The presence of radical scavengers blocks the
decrease in bcl-xL levels, m collapse,
cytochrome c release, and activation of caspase 3; in
contrast, the presence of glutathione synthesis inhibitors such as BSO
accentuated the effect. The incubation of fetal hepatocytes in the
presence of ter-butyl-hydroperoxide alone produces a decrease in
bcl-xL. These results indicate that during the apoptosis
mediated by TGF-ß in fetal hepatocytes, ROS may be responsible for
the decrease in bcl-xL mRNA levels that precedes the loss
of m, the release of cytochrome c, and
the activation of caspase 3, culminating in cell death.—Herrera, B.,
Alvarez, A. M., Sánchez, A., Fernández, M., Roncero,
C., Benito, M., Fabregat, I. Reactive oxygen species (ROS) mediates the
mitochondrial-dependent apoptosis induced by transforming growth factor
ß in fetal hepatocytes.
Key Words: cytochrome c • caspases • Bcl-x
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