Phenobarbital modulates the type of cell death by rat hepatocytes during deprivation of serum in vitro.

An immortal line of chemically altered rat hepatocytes was used to study the effects of the liver tumor promoter, phenobarbital (PB), on hepatocyte growth and viability in vitro. When the serum concentration in medium was changed from 10% to 0.5%, cell proliferation decreased and hepatocytes died. Death of the hepatocytes occurred after 2 days in low-serum medium. PB appeared to control the type of cell death that occurred. In the absence of PB in low-serum medium, most dead cells had morphological changes that are characteristic of necrosis as determined by both light and electron microscopy. In the presence of PB, the dead cells had alterations typical of apoptosis. Biochemical features of cell death in low-serum medium were also analyzed. DNA isolated from cells in low serum with PB showed nucleosome-length fragments after gel electrophoresis, whereas DNA from cells in low serum without PB appeared as randomly degraded fragments. Although proliferation of hepatocytes in low-serum decreased by 75%, the appearance of apoptosis in the presence of PB was associated with increased expression of the c-myc gene. Based on these observations, we conclude that PB can modulate the type of cell death that occurs after serum deprivation in this line of immortal rat hepatocytes. PB seemed to prevent necrotic cell death in low serum, and cells died through a gene-directed pathway of apoptosis.