C-myc is required for the G sub(0)/G sub(1)-S transition of primary hepatocytes stimulated with a deleted form of hepatocyte growth factor
Primary rat hepatocytes stimulated in vitro with the addition of a deleted form of hepatocyte growth factor (dHGF) enter the S-phase 48 h after addition of the growth factor. The c-myc gene is believed to play a role in a variety of cellular stages, such as proliferation, differentiation and cell de...
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| Published in: | Biochemical journal 1996-01, Vol.316 (3), p.879-886 |
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| Main Authors: | , |
| Format: | Article |
| Language: | eng |
| Online Access: | Get full text |
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| Summary: | Primary rat hepatocytes stimulated in vitro with the addition of a deleted form of hepatocyte growth factor (dHGF) enter the S-phase 48 h after addition of the growth factor. The c-myc gene is believed to play a role in a variety of cellular stages, such as proliferation, differentiation and cell death. In primary hepatocytes c-myc was expressed constitutively at both mRNA and protein levels, independently of the growth conditions. dHGF induced significant c-myc expression at times correlated with the long-lasting pre-S phase, and no induction was observed at the G sub(0)/G sub(1) traverse compared with the unstimulated hepatocytes. An antisense construct coding for all three exons of c-myc was imported into hepatocytes by using the transferrin receptor-mediated endocytosis methodology (transferrinfection). Expression of the antisense construct inhibited the biosynthesis of the c-Myc protein, however it did not interfere with the expression of c-met, encoding the receptor for HGF/dHGF. Continuous expression of the antisense construct inhibited entry of the hepatocytes into the S-phase. Regulated induction of the antisense c-myc by dexamethasone for up to 6 h in culture, did not interfere with the entry of hepatocytes into the S-phase. c-myc expression was shown to be required between 6 and 12 h in dHGF-stimulated hepatocytes, and inhibition of its expression during this time by the antisense myc construct did not allow these cells to enter the S-phase. Inhibition of c-myc biosynthesis between 24 and 48 h hours slightly affected the DNA synthetic response. It is proposed that the expression of c-Myc protein interferes with the 'priming' of hepatocytes to become responsive to growth-factor stimuli, or in the absence of such stimuli it interferes with the maintenance of a non-proliferating phenotype and subsequent in vitro de-differentiation. |
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| ISSN: | 0264-6021 1470-8728 |