Summary
The estrogen-responsive Leghorn strain M chicken hepatoma (LMH) cell line provides a model system for studying the estrogen-dependent, liver-specific expression of avian genes. Serum-free culture conditions have been established that allow expression of apolipoprotein B, very low density apolipoprotein II (apoVLDLII), serum albumin, and transferrin at levels detectable by Northern blot analysis. Regulation of apoVLDLII mRNA by estrogen occurred in an appropriate time-and dose-dependent manner in serum-free cultures of the LMH cells. The expression of apoVLDLII mRNA in serum-free culture was at least 100-fold higher than that expressed in cultures containing 10% serum. The level of estrogen receptors in LMH cells cultured with 10% serum was approximately 2000 receptors per cell, and in serum-free culture approximately 1000 receptors per cell. When these cells were transfected with estrogen receptor DNA and cultured in serum-free medium, apoVLDLII mRNA was decreased relative to that expressed in cells transfected with a control plasmid. These results indicate that when the LMH cells are cultured without serum, estrogen receptors are not the limiting factor for the expression of the apoVLDLII gene.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Beato, M. Gene regulation by steroid hormones. Cell 56:335–344; 1989
Binder, R.; MacDonald, C. C.; Burch, J. B. E., et al. Expression of endogenous and transfected apolipoprotein II genes in an estrogen responsive chicken liver cell line. Mol. Endocrinol. 4:201–208; 1990.
Burch, J. B. E.; Fischer, A. H. Chromatin studies reveal that an ERE is located far upstream of a vitellogenin gene and that a distal tissuespecific hypersensitive site is conserved for two coordinately regulated vitellogenin genes. Nucleic Acids Res. 18:4157–4165; 1990.
Burch, J. B. E.; Evans, M. I.; Friedman, T. M., et al. Two functional estrogen response elements are located upstream of the major chicken vitellogenin gene. Mol. Cell. Biol. 8:1123–1131; 1988.
Burton, K. A study of the conditions and mechanisms of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem. J. 62:315–323; 1956.
Chalepakis, G.; Arnemann, J.; Slater, E., et al. Differential gene activation by glucocorticoids and progestins through the hormone regulatory element of mouse mammary tumor virus. Cell 53:371–382; 1988.
Chambraud, B.; Berry, M.; Redeuilh, G., et al. Several regions of human estrogen receptor are involved in the formation of receptorheat shock protein 90 complexes. J. Biol. Chem. 265:20686–20691; 1990.
Cochet, M.; Perrin, F.; Gannon, F., et al. Cloning of an almost fulllength chicken conalbumin double-stranded cDNA. Nucleic Acids Res. 6:2435–2452; 1979.
Druege, P. M.; Klein-Hitpass, L.; Green, S., et al. Introduction of estrogen-responsiveness into mammalian cell lines. Nucleic Acids Res. 14:9329–9337; 1986.
Evans, M. I.; O’Malley, P. J.; Krust, A., et al. Developmental regulation of the estrogen receptor and the estrogen responsiveness of five yolk protein genes in the avian liver. Proc. Natl. Acad. Sci. USA 84:8493–8497; 1987.
Evans, M. I.; Silva, R.; Burch, J. B. E. Isolation of vitellogenin I and III cDNAs and the developmental regulation of five estrogen-responsive genes in the embryonic liver. Genes Dev. 2:116–124; 1988.
Gaub, M.-P.; Bellard, M.; Scheuer, I., et al. Activation of the ovalbumin gene by the estrogen receptor involves the fos-jun complex. Cell 63:1267–1276; 1990.
Gorski, J.; Gannon, F. Current models of steroid hormone action: a critique. Ann. Rev. Physiol. 38:425–450; 1976.
Graham, F. L.; Van Der Eb, A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–467; 1973.
Green, S.; Chambon, P. Nuclear receptors enhance our understanding of transcription regulation. Trends Genet. 4:309–314; 1988.
Katzenellenbogen, B. S.; Kendra, K. L.; Norman, M. J., et al. Proliferation, hormonal responsiveness, and estrogen receptor content of MCF-7 human breast cancer cells grown in the short-term and long-term absence of estrogens. Cancer Res. 47:4335–4360; 1987.
Kawaguchi, T.; Nomura, K.; Hirayama, Y., et al. Establishment and characterization of a chicken hepatocellular carcinoma cell line, LMH. Cancer Res. 47:4460–4464; 1987.
Kirchgessner, T. G.; Heinzmann, C.; Svenson, K. L., et al. Regulation of chicken apolipoprotein B: cloning, tissue distribution, and estrogen induction of mRNA. Gene 59:241–251; 1987.
Krust, A.; Green, S.; Argos, P., et al. The chicken oestrogen receptor sequence: homology with v-erb-A and the human oestrogen and glucocorticoid receptors. EMBO J. 5:891–897; 1986.
Medrano, E. E.; Resnicoff, M.; Cafferata, E. G. A., et al. Increased secretory activity and estradiol receptor expression are among other relevant aspects of MCF-7 human breast tumor cell growth which are expressed only in the absence of serum. Exp. Cell Res. 188:2–9; 1990.
Meinkoth, J.; Wahl, G. Hybridization of nucleic acids immobilized on solid supports. Anal. Biochem. 138:267–284; 1984.
Melton, D. A.; Krieg, P. A.; Rebagliati, M. R., et al. Efficientin vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 12:7035–7056; 1984.
Meyer, M.-E.; Gronemeyer, H.; Turcotte, B., et al. Steroid hormone receptors compete for factors that mediate their enhancer function. Cell 57:433–442; 1989.
Mukherjee, R.; Chambon, P. A single-stranded DNA-binding protein promotes the binding of the purified oestrogen receptor to its responsive element. Nucleic Acids Res. 18:5713–5716; 1990.
Picard, D.; Khursheed, B.; Garabedian, M. J., et al. Reduced levels of hsp90 compromise steroid receptor actionin vivo. Nature 348:166–168; 1990.
Pratt, W. B. Interaction of hsp-90 with steroid receptors: organizing some diverse observations and presenting the newest concepts. Mol. Cell. Endocrinol. 74:C69-C76; 1990.
Proter, A. A.; Wang, S.; Shelness, G. S., et al. Isolation and characterization of a cDNA clone specific for avian vitellogenin II. Nucleic Acids Res. 10:4935–4950; 1982.
Ptashne, M. How eukaryotic transcriptional activators work. Nature 335:683–689; 1988.
Pugh, B. F.; Tjian, R. Mechanism of transcriptional activation by SP1: evidence for coactivators. Cell 61:1187–1197; 1990.
Raynaud, J.-P.; Bouton, M.-M.; Gallet-Bourquin, D., et al. Comparative study of estrogen action. Mol. Pharmacol. 9:520–533; 1973.
Rentoumis, A.; Krishna, V.; Chatterjee, K., et al. Negative and positive transcriptional regulation by thyroid hormone receptor isoforms. Mol. Endocrinol. 4:1522–1531; 1990.
Sambrook, J.; Fritsch, E. F.; Maniatis, T. Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory; 1989.
Scatchard, G. The attraction of proteins for small molecules and ions. Annals NY Acad. Science 51:660–672; 1949.
Schule, R.; Rangarajan, P.; Kliewer, S., et al. Functional antagonism between oncoprotein c-Jun and the glucocorticoid receptor. Cell 62:1217–1226; 1990.
Weisz, A.; Cicatiello, L.; Persico, E., et al. Estrogen stimulates transcription of c-jun protooncogene. Mol. Endocrinol. 4:1041–1050; 1990.
Yamamoto, K. R.; Alberts, B. M. Steroid receptors: elements for modulation of eukaryotic transcription. Ann. Rev. Biochem. 45:721–746; 1976.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Berkowitz, E.A., Evans, M.I. Estrogen-dependent expression of the chicken very low density apolipoprotein II gene in serum-free cultures of LMH cells. In Vitro Cell Dev Biol - Animal 28, 391–396 (1992). https://doi.org/10.1007/BF02634041
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02634041