Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1988 Jan;8(1):203–209. doi: 10.1128/mcb.8.1.203

Regulation of rat liver maturation in vitro by glucocorticoids.

J Y Chou 1, Y J Wan 1, T Sakiyama 1
PMCID: PMC363102  PMID: 2447484

Abstract

The biochemistry of liver maturation was studied by using the RLA209-15 fetal rat hepatocyte line that is temperature sensitive for maintenance of the differentiated fetal liver phenotype. At 33 degrees C these cells were dedifferentiated; but at 40 degrees C they were phenotypically differentiated and, like normal fetal hepatocytes, synthesized moderate levels of albumin and transferrin, high levels of authentic (69,000 and 73,000 molecular weight) rat fetal alpha-fetoprotein (AFP), and low levels of a 65,000-molecular-weight variant AFP. Our results indicated that administration of glucocorticoid hormones to RLA209-15 cells at 40 degrees C induced a series of events associated with normal hepatocyte maturation; synthesis of fetal AFP was inhibited, whereas the synthesis of variant AFP, albumin, transferrin, tyrosine aminotransferase, and alpha 1-acid glycoprotein was induced. The variant AFP was produced by RLA209-15 cells at both temperatures and was encoded by an mRNA of 1.7 kilobases (kb). The fetal AFP was encoded by an mRNA of 2.2 kb. Normal adult rat liver contained three AFP mRNAs of 2.2 (minor), 1.7, and 1.5 kb. The 1.7-kb adult liver AFP mRNA comigrated with the RNA found in RLA209-15 cells, and both directed the synthesis of a 50,000-molecular-weight precursor polypeptide of the variant AFP. Administration of glucocorticoids to RLA209-15 cells grown at 33 degrees C stimulated synthesis of both the fetal and variant AFPs, but the levels of the 2.2-kb AFP mRNA were preferentially increased. RLA209-15 cells contained two glucocorticoid receptor mRNAs of 6.8 and 4.5 kb. The glucocorticoid-mediated maturation described above was blocked by the antiglucocorticoid RU486.

Full text

PDF
203

Images in this article

205Image
on p.205
206Image
on p.206
206Image
on p.206
207Image
on p.207

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abelev G. I. Alpha-fetoprotein in ontogenesis and its association with malignant tumors. Adv Cancer Res. 1971;14:295–358. doi: 10.1016/s0065-230x(08)60523-0. [DOI] [PubMed] [Google Scholar]
  2. Baumann H., Maquat L. E. Localization of DNA sequences involved in dexamethasone-dependent expression of the rat alpha 1-acid glycoprotein gene. Mol Cell Biol. 1986 Jul;6(7):2551–2561. doi: 10.1128/mcb.6.7.2551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  4. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  5. Chou J. Y., Ito F., Evans G., Chiu J. F., Feldman M. alpha-Fetoprotein biosynthesis and hepatocellular differentiation. Biochem Biophys Res Commun. 1982 Oct 29;108(4):1524–1530. doi: 10.1016/s0006-291x(82)80080-6. [DOI] [PubMed] [Google Scholar]
  6. Chou J. Y., Savitz A. J. alpha-Fetoprotein synthesis in transformed fetal rat liver cells. Biochem Biophys Res Commun. 1986 Mar 28;135(3):844–851. doi: 10.1016/0006-291x(86)91005-3. [DOI] [PubMed] [Google Scholar]
  7. Chou J. Y., Schlegel-Haueter S. E. Study of liver differentiation in vitro. J Cell Biol. 1981 May;89(2):216–222. doi: 10.1083/jcb.89.2.216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chou J. Y. Temperature-sensitive adult liver cell line dependent on glucocorticoid for differentiation. Mol Cell Biol. 1983 Jun;3(6):1013–1020. doi: 10.1128/mcb.3.6.1013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Commer P., Schwartz C., Tracy S., Tamaoki T., Chiu J. F. Dexamethasone inhibits alpha-fetoprotein gene expression in developing mouse liver. Biochem Biophys Res Commun. 1979 Aug 28;89(4):1294–1299. doi: 10.1016/0006-291x(79)92149-1. [DOI] [PubMed] [Google Scholar]
  10. Ernest M. J., Chen C. L., Feigelson P. Induction of tyrosine aminotransferase synthesis in isolated liver cell suspensions. Absolute dependence of induction on glucocorticoids and glucagon or cyclic AMP. J Biol Chem. 1977 Oct 10;252(19):6783–6791. [PubMed] [Google Scholar]
  11. Gal A., Nahon J. L., Sala-Trepat J. M. Detection of rare mRNA species in a complex RNA population by blot hybridization techniques: a comparative survey. Anal Biochem. 1983 Jul 1;132(1):190–194. doi: 10.1016/0003-2697(83)90446-3. [DOI] [PubMed] [Google Scholar]
  12. Hager L. J., McKnight G. S., Palmiter R. D. Glucocorticoid induction of egg white mRNAs in chick oviduct. J Biol Chem. 1980 Aug 25;255(16):7796–7800. [PubMed] [Google Scholar]
  13. Jagodzinski L. L., Sargent T. D., Yang M., Glackin C., Bonner J. Sequence homology between RNAs encoding rat alpha-fetoprotein and rat serum albumin. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3521–3525. doi: 10.1073/pnas.78.6.3521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jantzen H. M., Strähle U., Gloss B., Stewart F., Schmid W., Boshart M., Miksicek R., Schütz G. Cooperativity of glucocorticoid response elements located far upstream of the tyrosine aminotransferase gene. Cell. 1987 Apr 10;49(1):29–38. doi: 10.1016/0092-8674(87)90752-5. [DOI] [PubMed] [Google Scholar]
  15. Jung-Testas I., Baulieu E. E. Inhibition of glucocorticosteroid action in cultured L-929 mouse fibroblasts by RU 486, a new anti-glucocorticosteroid of high affinity for the glucocorticosteroid receptor. Exp Cell Res. 1983 Aug;147(1):177–182. doi: 10.1016/0014-4827(83)90282-3. [DOI] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  18. Levin M. J., Tuil D., Uzan G., Dreyfus J. C., Kahn A. Expression of the transferrin gene during development of non-hepatic tissues: high level of transferrin mRNA in fetal muscle and adult brain. Biochem Biophys Res Commun. 1984 Jul 18;122(1):212–217. doi: 10.1016/0006-291x(84)90461-3. [DOI] [PubMed] [Google Scholar]
  19. Martin R. G., Chou J. Y. Simian virus 40 functions required for the establishment and maintenance of malignant transformation. J Virol. 1975 Mar;15(3):599–612. doi: 10.1128/jvi.15.3.599-612.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Miesfeld R., Rusconi S., Godowski P. J., Maler B. A., Okret S., Wikström A. C., Gustafsson J. A., Yamamoto K. R. Genetic complementation of a glucocorticoid receptor deficiency by expression of cloned receptor cDNA. Cell. 1986 Aug 1;46(3):389–399. doi: 10.1016/0092-8674(86)90659-8. [DOI] [PubMed] [Google Scholar]
  21. Moguilewsky M., Philibert D. RU 38486: potent antiglucocorticoid activity correlated with strong binding to the cytosolic glucocorticoid receptor followed by an impaired activation. J Steroid Biochem. 1984 Jan;20(1):271–276. doi: 10.1016/0022-4731(84)90216-4. [DOI] [PubMed] [Google Scholar]
  22. Nawa K., Nakamura T., Kumatori A., Noda C., Ichihara A. Glucocorticoid-dependent expression of the albumin gene in adult rat hepatocytes. J Biol Chem. 1986 Dec 25;261(36):16883–16888. [PubMed] [Google Scholar]
  23. Nickol J. M., Lee K. L., Kenney F. T. Changes in hepatic levels of tyrosine aminotransferase messenger RNA during induction by hydrocortisone. J Biol Chem. 1978 Jun 10;253(11):4009–4015. [PubMed] [Google Scholar]
  24. Parsa I., Flancbaum L. Long term organ culture of rat liver rudiment in a synthetic medium: morphological and biochemical development. Dev Biol. 1975 Sep;46(1):120–131. doi: 10.1016/0012-1606(75)90091-3. [DOI] [PubMed] [Google Scholar]
  25. Reinke R., Feigelson P. Rat alpha 1-acid glycoprotein. Gene sequence and regulation by glucocorticoids in transfected L-cells. J Biol Chem. 1985 Apr 10;260(7):4397–4403. [PubMed] [Google Scholar]
  26. Ricca G. A., Taylor J. M. Nucleotide sequence of rat alpha 1-acid glycoprotein messenger RNA. J Biol Chem. 1981 Nov 10;256(21):11199–11202. [PubMed] [Google Scholar]
  27. Sargent T. D., Wu J. R., Sala-Trepat J. M., Wallace R. B., Reyes A. A., Bonner J. The rat serum albumin gene: analysis of cloned sequences. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3256–3260. doi: 10.1073/pnas.76.7.3256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Scherer G., Schmid W., Strange C. M., Röwekamp W., Schütz G. Isolation of cDNA clones coding for rat tyrosine aminotransferase. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7205–7208. doi: 10.1073/pnas.79.23.7205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sell S., Skelly H., Leffert H. L., Muller-Eberhard U., Kida S. Relationship of the biosynthesis of alpha-fetoprotein, albumin, hemopexin, and haptoglobin to the growth state of fetal rat hepatocyte cultures. Ann N Y Acad Sci. 1975 Aug 22;259:45–58. doi: 10.1111/j.1749-6632.1975.tb25401.x. [DOI] [PubMed] [Google Scholar]
  30. Short J. M., Wynshaw-Boris A., Short H. P., Hanson R. W. Characterization of the phosphoenolpyruvate carboxykinase (GTP) promoter-regulatory region. II. Identification of cAMP and glucocorticoid regulatory domains. J Biol Chem. 1986 Jul 25;261(21):9721–9726. [PubMed] [Google Scholar]
  31. Thomas T., Schreiber G. Acute-phase response of plasma protein synthesis during experimental inflammation in neonatal rats. Inflammation. 1985 Mar;9(1):1–7. doi: 10.1007/BF00915406. [DOI] [PubMed] [Google Scholar]
  32. Tilghman S. M., Belayew A. Transcriptional control of the murine albumin/alpha-fetoprotein locus during development. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5254–5257. doi: 10.1073/pnas.79.17.5254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Turcotte B., Guertin M., Chevrette M., Bélanger L. Rat alpha 1-fetoprotein messenger RNA: 5'-end sequence and glucocorticoid-suppressed liver transcription in an improved nuclear run-off assay. Nucleic Acids Res. 1985 Apr 11;13(7):2387–2398. doi: 10.1093/nar/13.7.2387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yamamoto K. R. Steroid receptor regulated transcription of specific genes and gene networks. Annu Rev Genet. 1985;19:209–252. doi: 10.1146/annurev.ge.19.120185.001233. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES