Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1984 Apr;4(4):712–721. doi: 10.1128/mcb.4.4.712

Hormonal regulation of the transformation phenotype in simian virus 40-transformed rat embryonic preadipose cell lines.

S Yasumoto
PMCID: PMC368787  PMID: 6325892

Abstract

The regulation of transformed phenotypes was studied in newly isolated preadipose cell lines which were established after infection with simian virus 40 tsA58 dl2009. The clonal cell lines isolated exhibited most of the characteristics typical of transformed cells. The transformants, however, were able to differentiate into adipocytes in the presence of low calf serum (0.5%) and a combination of several hormones, including hydrocortisone and insulin. Treatment with insulin alone stimulated the growth of these cells but did not induce lipid accumulation without added hydrocortisone. The effect of hydrocortisone was accompanied by a restoration of growth control in the transformants after they reached high cell density. The blot hybridization analysis of cellular DNAs digested by restriction enzymes revealed that simian virus 40 genomes were integrated at multiple separate sites at which a head-to-tail oligomeric insertion took place. Large T antigen was synthesized in growing cells but was regulated at high cell density when cells were committed to differentiate by glucocorticoids. These results suggest that the glucocorticoid hydrocortisone is capable of restoring growth regulation at high cell densities to simian virus 40-transformed preadipose cell lines.

Full text

PDF
712

Images in this article

Selected References

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

  1. Adams S. L., Sobel M. E., Howard B. H., Olden K., Yamada K. M., de Crombrugghe B., Pastan I. Levels of translatable mRNAs for cell surface protein, collagen precursors, and two membrane proteins are altered in Rous sarcoma virus-transformed chick embryo fibroblasts. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3399–3403. doi: 10.1073/pnas.74.8.3399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ambesi-Impiombato F. S., Parks L. A., Coon H. G. Culture of hormone-dependent functional epithelial cells from rat thyroids. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3455–3459. doi: 10.1073/pnas.77.6.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Banks-Schlegel S. P., Howley P. M. Differentiation of human epidermal cells transformed by SV40. J Cell Biol. 1983 Feb;96(2):330–337. doi: 10.1083/jcb.96.2.330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bartholomew J. C., Yokota H., Ross P. Effect of serum on the growth of Balb oT3 A31 mouse fibroblasts and an SV40-transformed derivative. J Cell Physiol. 1976 Jul;88(3):277–286. doi: 10.1002/jcp.1040880303. [DOI] [PubMed] [Google Scholar]
  5. Chou J. Y. Establishment of clonal human placental cells synthesizing human choriogonadotropin. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1854–1858. doi: 10.1073/pnas.75.4.1854. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chou J. Y. Human placental cells transformed by tsA mutants of simian virus 40: a model system for the study of placental functions. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1409–1413. doi: 10.1073/pnas.75.3.1409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Collins J. J., Black P. H., Strosberg A. D., Haber E., Bloch K. J. Transformation by simian virus 40 of spleen cells from a hyperimmune rabbit: evidence for synthesis of immunoglobulin by the transformed cells. Proc Natl Acad Sci U S A. 1974 Feb;71(2):260–262. doi: 10.1073/pnas.71.2.260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Darmon M., Serrero G., Rizzino A., Sato G. Isolation of myoblastic, fibro-adipogenic, and fibroblastic clonal cell lines from a common precursor and study of their requirements for growth and differentiation. Exp Cell Res. 1981 Apr;132(2):313–327. doi: 10.1016/0014-4827(81)90107-5. [DOI] [PubMed] [Google Scholar]
  9. Dubrow R., Riddle V. G., Pardee A. B. Different responses to drugs and serum of cells transformed by various means. Cancer Res. 1979 Jul;39(7 Pt 1):2718–2726. [PubMed] [Google Scholar]
  10. Green H., Kehinde O. An established preadipose cell line and its differentiation in culture. II. Factors affecting the adipose conversion. Cell. 1975 May;5(1):19–27. doi: 10.1016/0092-8674(75)90087-2. [DOI] [PubMed] [Google Scholar]
  11. Green H., Meuth M. An established pre-adipose cell line and its differentiation in culture. Cell. 1974 Oct;3(2):127–133. doi: 10.1016/0092-8674(74)90116-0. [DOI] [PubMed] [Google Scholar]
  12. Greenberger J. S. Sensitivity of corticosteroid-dependent insulin-resistant lipogenesis in marrow preadipocytes of obese-diabetic (db/db) mice. Nature. 1978 Oct 26;275(5682):752–754. doi: 10.1038/275752a0. [DOI] [PubMed] [Google Scholar]
  13. Kodama H. A., Amagai Y., Koyama H., Kasai S. Hormonal responsiveness of a preadipose cell line derived from newborn mouse calvaria. J Cell Physiol. 1982 Jul;112(1):83–88. doi: 10.1002/jcp.1041120113. [DOI] [PubMed] [Google Scholar]
  14. Krawisz B. R., Scott R. E. Coupling of proadipocyte growth arrest and differentiation. I. Induction by heparinized medium containing human plasma. J Cell Biol. 1982 Aug;94(2):394–399. doi: 10.1083/jcb.94.2.394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kuri-Harcuch W., Green H. Adipose conversion of 3T3 cells depends on a serum factor. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6107–6109. doi: 10.1073/pnas.75.12.6107. [DOI] [PMC free article] [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. Lanotte M., Scott D., Dexter T. M., Allen T. D. Clonal preadipocyte cell lines with different phenotypes derived from murine marrow stroma: factors influencing growth and adipogenesis in vitro. J Cell Physiol. 1982 May;111(2):177–186. doi: 10.1002/jcp.1041110209. [DOI] [PubMed] [Google Scholar]
  18. Linzer D. I., Levine A. J. Characterization of a 54K dalton cellular SV40 tumor antigen present in SV40-transformed cells and uninfected embryonal carcinoma cells. Cell. 1979 May;17(1):43–52. doi: 10.1016/0092-8674(79)90293-9. [DOI] [PubMed] [Google Scholar]
  19. MACPHERSON I., MONTAGNIER L. AGAR SUSPENSION CULTURE FOR THE SELECTIVE ASSAY OF CELLS TRANSFORMED BY POLYOMA VIRUS. Virology. 1964 Jun;23:291–294. doi: 10.1016/0042-6822(64)90301-0. [DOI] [PubMed] [Google Scholar]
  20. Mackall J. C., Student A. K., Polakis S. E., Lane M. D. Induction of lipogenesis during differentiation in a "preadipocyte" cell line. J Biol Chem. 1976 Oct 25;251(20):6462–6464. [PubMed] [Google Scholar]
  21. Magun B. E., Thompson R. L., Gerner E. W. Regulation of DNA replication by serum and the transforming function in cultured rat fibroblasts transformed by Rous sarcoma virus. J Cell Physiol. 1979 May;99(2):207–216. doi: 10.1002/jcp.1040990207. [DOI] [PubMed] [Google Scholar]
  22. Martin R. G., Setlow V. P., Chepelinsky A. B., Seif R., Lewis A. M., Jr, Scher C. D., Stiles C. D., Avila J. Roles of the T antigens in transformation by SV40. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 1):311–324. doi: 10.1101/sqb.1980.044.01.036. [DOI] [PubMed] [Google Scholar]
  23. Martin R. G., Stein S. Resting state in normal and simian virus 40 transformed Chinese hamster lung cells. Proc Natl Acad Sci U S A. 1976 May;73(5):1655–1659. doi: 10.1073/pnas.73.5.1655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Négrel R., Grimaldi P., Ailhaud G. Establishment of preadipocyte clonal line from epididymal fat pad of ob/ob mouse that responds to insulin and to lipolytic hormones. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6054–6058. doi: 10.1073/pnas.75.12.6054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. O'Neill F. J. Differential in vitro growth properties of cells transformed by DNA and RNA tumor viruses. Exp Cell Res. 1978 Dec;117(2):393–401. doi: 10.1016/0014-4827(78)90152-0. [DOI] [PubMed] [Google Scholar]
  26. Oliver N., Newby R. F., Furcht L. T., Bourgeois S. Regulation of fibronectin biosynthesis by glucocorticoids in human fibrosarcoma cells and normal fibroblasts. Cell. 1983 May;33(1):287–296. doi: 10.1016/0092-8674(83)90357-4. [DOI] [PubMed] [Google Scholar]
  27. Paul D., Henahan M., Walter S. Changes in growth control and growth requirements associated with neoplastic transformation in vitro. J Natl Cancer Inst. 1974 Nov;53(5):1499–1503. doi: 10.1093/jnci/53.5.1499. [DOI] [PubMed] [Google Scholar]
  28. Paul D. Quiescent SV40 virus transformed 3T3 cells in culture. Biochem Biophys Res Commun. 1973 Aug 6;53(3):745–753. doi: 10.1016/0006-291x(73)90156-3. [DOI] [PubMed] [Google Scholar]
  29. Pollack Y., Stein R., Razin A., Cedar H. Methylation of foreign DNA sequences in eukaryotic cells. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6463–6467. doi: 10.1073/pnas.77.11.6463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  31. Risser R., Pollack R. A nonselective analysis of SV40 transformation of mouse 3T3 cells. Virology. 1974 Jun;59(2):477–489. doi: 10.1016/0042-6822(74)90457-7. [DOI] [PubMed] [Google Scholar]
  32. Risser R., Pollack R. Factors affecting the frequency of transformation of rat embryo cells by simian virus 40. Virology. 1979 Jan 15;92(1):82–90. doi: 10.1016/0042-6822(79)90216-2. [DOI] [PubMed] [Google Scholar]
  33. Roberts R. J. Restriction and modification enzymes and their recognition sequences. Nucleic Acids Res. 1980 Jan 11;8(1):r63–r80. doi: 10.1093/nar/8.1.197-d. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Robinson C. C., Lehman J. M. Simian virus 40 A gene function: DNA content analysis of Chinese hamster cells transformed by an early temperature-sensitive virus mutant. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4389–4393. doi: 10.1073/pnas.75.9.4389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rubin C. S., Hirsch A., Fung C., Rosen O. M. Development of hormone receptors and hormonal responsiveness in vitro. Insulin receptors and insulin sensitivity in the preadipocyte and adipocyte forms of 3T3-L1 cells. J Biol Chem. 1978 Oct 25;253(20):7570–7578. [PubMed] [Google Scholar]
  36. Russell T. R., Ho R. Conversion of 3T3 fibroblasts into adipose cells: triggering of differentiation by prostaglandin F2alpha and 1-methyl-3-isobutyl xanthine. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4516–4520. doi: 10.1073/pnas.73.12.4516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sager R., Kovac P. Pre-adipocyte determination either by insulin or by 5-azacytidine. Proc Natl Acad Sci U S A. 1982 Jan;79(2):480–484. doi: 10.1073/pnas.79.2.480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schlegel-Haueter S. E., Schlegel W., Chou J. Y. Establishment of a fetal rat liver cell line that retains differentiated liver functions. Proc Natl Acad Sci U S A. 1980 May;77(5):2731–2734. doi: 10.1073/pnas.77.5.2731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Scott R. E., Florine D. L., Wille J. J., Jr, Yun K. Coupling of growth arrest and differentiation at a distinct state in the G1 phase of the cell cycle: GD. Proc Natl Acad Sci U S A. 1982 Feb;79(3):845–849. doi: 10.1073/pnas.79.3.845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Scott R. E., Hoerl B. J., Wille J. J., Jr, Florine D. L., Krawisz B. R., Yun K. Coupling of proadipocyte growth arrest and differentiation. II. A cell cycle model for the physiological control of cell proliferation. J Cell Biol. 1982 Aug;94(2):400–405. doi: 10.1083/jcb.94.2.400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  42. Steinberg M. L., Defendi V. Altered pattern of growth and differentiation in human keratinocytes infected by simian virus 40. Proc Natl Acad Sci U S A. 1979 Feb;76(2):801–805. doi: 10.1073/pnas.76.2.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Strosberg A. D., Collins J. J., Black P. H., Malamud D., Wilbert S., Bloch K. J., Haber E. Transformation by simian virus 40 of spleen cells from a hyperimmune rabbit: demonstration of production of specific antibody to the immunizing antigen. Proc Natl Acad Sci U S A. 1974 Feb;71(2):263–264. doi: 10.1073/pnas.71.2.263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Takaoka T., Yasumoto S., Katsuta H. A simple method for the cultivation of rat liver cells. Jpn J Exp Med. 1975 Oct;45(5):317–326. [PubMed] [Google Scholar]
  45. Taylor S. M., Jones P. A. Multiple new phenotypes induced in 10T1/2 and 3T3 cells treated with 5-azacytidine. Cell. 1979 Aug;17(4):771–779. doi: 10.1016/0092-8674(79)90317-9. [DOI] [PubMed] [Google Scholar]
  46. Tegtmeyer P., Schwartz M., Collins J. K., Rundell K. Regulation of tumor antigen synthesis by simain virus 40 gene A. J Virol. 1975 Jul;16(1):168–178. doi: 10.1128/jvi.16.1.168-178.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. 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]
  48. Wille J. J., Jr, Maercklein P. B., Scott R. E. Neoplastic transformation and defective control of cell proliferation and differentiation. Cancer Res. 1982 Dec;42(12):5139–5146. [PubMed] [Google Scholar]
  49. Yasumoto S., Kondo S., Kato Y. Growth and differentiation of primary chick embryonic chondrocytes on agar plates. Jpn J Exp Med. 1980 Jun;50(3):221–224. [PubMed] [Google Scholar]
  50. von der Mark K., Gauss V., von der Mark H., Müller P. Relationship between cell shape and type of collagen synthesised as chondrocytes lose their cartilage phenotype in culture. Nature. 1977 Jun 9;267(5611):531–532. doi: 10.1038/267531a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES