Liver-specific RNA metabolism in hepatocytes in long-term culture
The authors have examined the transcription of liver-specific and housekeeping genes in rat hepatocytes maintained in culture for up to 40 days in chemically defined medium. RNA extracted from these cells was analyzed using northern and dot blot methods. RNA in nuclei extracted from parallel cultures was analyzed by nascent chain extension experiments. Autoradiography of northern and dot blots indicated that in vitro hepatocyte gene expression relative to in vivo liver gene expression can be divided into four classes: (1) liver-specific genes that show loss of expression with time in culture; (2) liver-specific genes that are expressed for up to 40 days in culture at levels approximately 30-100% of those found in liver, e.g., albumin; (3) housekeeping genes that are expressed for up to 40 days in culture at levels approximately equivalent to those found in liver; and (4) housekeeping genes that are expressed at higher levels in cells in culture than liver. The results of nascent chain extension experiments indicated that the rate of synthesis of albumin message was many fold lower in hepatocytes than in liver, whereas rates of synthesis of messages for housekeeping genes were equal to or slightly higher in nuclei from hepatocytes in culture than in nuclei extracted from liver. Molecular analyses of nuclear RNA suggest that the high levels of albumin mRNA in cultured cells are due to an increased stability probably in the cytoplasm.
- Research Organization:
- Pennsylvania State Univ. College of Medicine, New York, NY
- OSTI ID:
- 5232955
- Report Number(s):
- CONF-8606151-
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
- Country of Publication:
- United States
- Language:
- English
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