Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Mar;79(6):1800–1804. doi: 10.1073/pnas.79.6.1800

Tumor promoters increase the synthesis of a 32,000-dalton protein in BALB/c 3T3 cells.

T Hiwasa, S Fujimura, S Sakiyama
PMCID: PMC346068  PMID: 6952230

Abstract

The effect of 12-O-tetradecanoylphorbol 13-acetate (TPA), a potent tumor promoter, on the synthesis of proteins in BALB/c 3T3 mouse fibroblasts has been studied. When [35S]methionine-labeled proteins synthesized after the addition of TPA were analyzed by one- or two-dimensional polyacrylamide gel electrophoresis, the increased synthesis of a 32,000-dalton protein (designated p32) was noted as one of the earliest changes. The synthesis of p32 increased approximately 2-fold within 2 hr at a promoter concentration of 20 ng/ml. Phorbol 12,13-didecanoate, another potent tumor promoter, showed the same effect as TPA, whereas 4-O-methyl-TPA, which has little promoting activity, did not enhance the synthesis of p32 at the same concentration but did effect a slight increase at 1 microgram/ml. p32 differed immunologically from a major excreted protein discovered by Gottesman [Gottesman, M. M. (1978) Proc. Natl. Acad. Sci. USA 75, 2767-2771]. The increased rate of the synthesis of p32 was considered to be regulated at the transcriptional level, because the increase in synthesis of p32 was inhibited by actinomycin D, and TPA-treated cells were shown to contain a higher level of translatable mRNA coding for p32 than do control cells. A possible relationship between the increase in p32 synthesis and transformation is discussed.

Full text

PDF
1800

Images in this article

1802Image
on p.1802
1802Image
on p.1802
1802Image
on p.1802
1802Image
on p.1802
1802Image
on p.1802
1803Image
on p.1803
1803Image
on p.1803
1803Image
on p.1803

Selected References

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

  1. Aaronson S. A., Todaro G. J. Basis for the acquisition of malignant potential by mouse cells cultivated in vitro. Science. 1968 Nov 29;162(3857):1024–1026. doi: 10.1126/science.162.3857.1024. [DOI] [PubMed] [Google Scholar]
  2. Aaronson S. A., Weaver C. A. Characterization of murine sarcoma virus (Kirsten) transformation of mouse and human cells. J Gen Virol. 1971 Nov;13(2):245–252. doi: 10.1099/0022-1317-13-2-245. [DOI] [PubMed] [Google Scholar]
  3. Balmain A. The synthesis of specific proteins in adult mouse epidermis during phases of proliferation and differentiation induced by the tumor promoter TPA, and in basal and differentiating layers of neonatal mouse epidermis. J Invest Dermatol. 1976 Aug;67(2):246–253. doi: 10.1111/1523-1747.ep12513451. [DOI] [PubMed] [Google Scholar]
  4. Belman S., Troll W. Phorbol-12-myristate-13-acetate effect on cyclic adenosine 3',5'-monophosphate levels in mouse skin and inhibition of phorbol-myristate-acetate-promoted tumorigenesis by theophylline. Cancer Res. 1974 Dec;34(12):3446–3455. [PubMed] [Google Scholar]
  5. Blumberg P. M., Driedger P. E., Rossow P. W. Effect of a phorbol ester on a transformation-sensitive surface protein of chick fibroblasts. Nature. 1976 Dec 2;264(5585):446–447. doi: 10.1038/264446a0. [DOI] [PubMed] [Google Scholar]
  6. Boreiko C., Mondal S., Narayan K. S., Heidelberger C. Effect of 12-O-tetradecanoylphorbol-13-acetate on the morphology and growth of C3H/10T1/2 mouse embryo cells. Cancer Res. 1980 Dec;40(12):4709–4716. [PubMed] [Google Scholar]
  7. Cabral F., Gottesman M. M., Yuspa S. H. Induction of specific protein synthesis by phorbol esters in mouse epidermal cell culture. Cancer Res. 1981 Jun;41(6):2025–2031. [PubMed] [Google Scholar]
  8. Cheung W. Y. Calmodulin plays a pivotal role in cellular regulation. Science. 1980 Jan 4;207(4426):19–27. doi: 10.1126/science.6243188. [DOI] [PubMed] [Google Scholar]
  9. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  10. Diamond L., O'Brien S., Donaldson C., Shimizu Y. Growth stimulation of human diploid fibro-blasts by the tumor promoter, 12-0-tetradecanoly-phorbol-13-acetate. Int J Cancer. 1974 May 15;13(5):721–730. doi: 10.1002/ijc.2910130516. [DOI] [PubMed] [Google Scholar]
  11. Driedger P. E., Blumberg P. M. The effect of phorbol diesters on chicken embryo fibroblasts. Cancer Res. 1977 Sep;37(9):3257–3265. [PubMed] [Google Scholar]
  12. Flower R., Gryglewski R., Herbaczyńska-Cedro K., Vane J. R. Effects of anti-inflammatory drugs on prostaglandin biosynthesis. Nat New Biol. 1972 Jul 26;238(82):104–106. doi: 10.1038/newbio238104a0. [DOI] [PubMed] [Google Scholar]
  13. Froscio M., Guy G. R., Murray A. W. Calmodulin inhibitors modify cell surface changes triggered by a tumor promoter. Biochem Biophys Res Commun. 1981 Feb 12;98(3):829–835. doi: 10.1016/0006-291x(81)91186-4. [DOI] [PubMed] [Google Scholar]
  14. Fürstenberger G., Marks F. Early prostaglandin E synthesis is an obligatory event in the induction of cell proliferation in mouse epidermis in vivo by the phorbol ester TPA. Biochem Biophys Res Commun. 1980 Feb 12;92(3):749–756. doi: 10.1016/0006-291x(80)90767-6. [DOI] [PubMed] [Google Scholar]
  15. Fürstenberger G., Marks F. Indomethacin inhibition of cell proliferation induced by the phorbolester TPA is reversed by prostaglandin E2 in mouse epidermis in vivo. Biochem Biophys Res Commun. 1978 Oct 30;84(4):1103–1111. doi: 10.1016/0006-291x(78)91697-2. [DOI] [PubMed] [Google Scholar]
  16. Gottesman M. M., Cabral F. Purification and characterization of a transformation-dependent protein secreted by cultured murine fibroblasts. Biochemistry. 1981 Mar 17;20(6):1659–1665. doi: 10.1021/bi00509a039. [DOI] [PubMed] [Google Scholar]
  17. Gottesman M. M., Sobel M. E. Tumor promoters and Kirsten sarcoma virus increase synthesis of a secreted glycoprotein by regulating levels of translatable mRNA. Cell. 1980 Feb;19(2):449–455. doi: 10.1016/0092-8674(80)90519-x. [DOI] [PubMed] [Google Scholar]
  18. Gottesman M. M. Transformation-dependent secretion of a low molecular weight protein by murine fibroblasts. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2767–2771. doi: 10.1073/pnas.75.6.2767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Grimm W., Marks F. Effect of tumor-promoting phorbol esters on the normal and the isoproterenol-elevated level of adenosine 3',5'-cyclic monophosphate in mouse epidermis in vivo. Cancer Res. 1974 Nov;34(11):3128–3134. [PubMed] [Google Scholar]
  20. Hennings H., Bowden G. T., Boutwell R. K. The effect of croton oil pretreatment on skin tumor initiation in mice. Cancer Res. 1969 Oct;29(10):1773–1780. [PubMed] [Google Scholar]
  21. Hynes R. O. Alteration of cell-surface proteins by viral transformation and by proteolysis. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3170–3174. doi: 10.1073/pnas.70.11.3170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kennedy A. R., Mondal S., Heidelberger C., Little J. B. Enhancement of X-ray transformation by 12-O-tetradecanoyl-phorbol-13-acetate in a cloned line of C3H mouse embryo cells. Cancer Res. 1978 Feb;38(2):439–443. [PubMed] [Google Scholar]
  23. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  24. 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]
  25. Laskin J. D., Mufson R. A., Piccinini L., Engelhardt D. L., Weinstein I. B. Effects of the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate on newly synthesized proteins in mouse epidermis. Cell. 1981 Aug;25(2):441–449. doi: 10.1016/0092-8674(81)90062-3. [DOI] [PubMed] [Google Scholar]
  26. Lasne C., Gentil A., Chouroulinkov I. Two-stage malignant transformation of rat fibroblasts in tissue culture. Nature. 1974 Feb 15;247(5441):490–491. doi: 10.1038/247490a0. [DOI] [PubMed] [Google Scholar]
  27. Levine L., Hassid A. Effects of phorbol-12,13-diesters on prostaglandin production and phospholipase activity in canine kidney (MDCK) cells. Biochem Biophys Res Commun. 1977 Nov 21;79(2):477–484. doi: 10.1016/0006-291x(77)90182-6. [DOI] [PubMed] [Google Scholar]
  28. Marks F., Fürstenberger G., Kownatzki E. Prostaglandin E-mediated mitogenic stimulation of mouse epidermis in vivo by divalent cation ionophore A 23187 and by tumor promoter 12-O-tetradecanoylphorbol-13-acetate. Cancer Res. 1981 Feb;41(2):696–702. [PubMed] [Google Scholar]
  29. Mondal S., Brankow D. W., Heidelberger C. Two-stage chemical oncogenesis in cultures of C3H/10T1/2 cells. Cancer Res. 1976 Jul;36(7 Pt 1):2254–2260. [PubMed] [Google Scholar]
  30. Mondal S., Heidelberger C. Transformation of C3H/10T1/2CL8 mouse embryo fibroblasts by ultraviolet irradiation and a phorbol ester. Nature. 1976 Apr 22;260(5553):710–711. doi: 10.1038/260710a0. [DOI] [PubMed] [Google Scholar]
  31. O'Brien T. G., Simsiman R. C., Boutwell R. K. Induction of the polyamine-biosynthetic enzymes in mouse epidermis and their specificity for tumor promotion. Cancer Res. 1975 Sep;35(9):2426–2433. [PubMed] [Google Scholar]
  32. O'Brien T. G., Simsiman R. C., Boutwell R. K. Induction of the polyamine-biosynthetic enzymes in mouse epidermis by tumor-promoting agents. Cancer Res. 1975 Jul;35(7):1662–1670. [PubMed] [Google Scholar]
  33. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  34. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  35. Perchellet J. P., Boutwell R. K. Effects of 3-isobutyl-1-methylxanthine and cyclic nucleotides on 12-O-tetradecanoylphorbol-13-acetate-induced ornithine decarboxylase activity in mouse epidermis in vivo. Cancer Res. 1981 Oct;41(10):3918–3926. [PubMed] [Google Scholar]
  36. Rifkin D. B., Crowe R. M., Pollack R. Tumor promoters induce changes in the chick embryo fibroblast cytoskeleton. Cell. 1979 Oct;18(2):361–368. doi: 10.1016/0092-8674(79)90055-2. [DOI] [PubMed] [Google Scholar]
  37. Sakiyama S., Fujimura S., Sakiyama H. Absence of gamma-actin expression in the mouse fibroblast cell line, L. J Biol Chem. 1981 Jan 10;256(1):31–33. [PubMed] [Google Scholar]
  38. Schimke R. T., Palacios R., Sullivan D., Kiely M. L., Gonzales C., Taylor J. M. Immunoadsorption of ovalbumin synthesizing polysosmes and partial purification of ovalbumin messenger RNA. Methods Enzymol. 1974;30:631–648. doi: 10.1016/0076-6879(74)30061-4. [DOI] [PubMed] [Google Scholar]
  39. Scribner J. D., Slaga T. J. Influence of nonsteroid anti-inflammatory agents on protein synthesis and hyperplasia caused by a tumor promoter. J Natl Cancer Inst. 1974 Jun;52(6):1865–1868. doi: 10.1093/jnci/52.6.1865. [DOI] [PubMed] [Google Scholar]
  40. Sivak A., Van Duuren B. L. RNA synthesis induction in cell culture by a tumor promoter. Cancer Res. 1970 Apr;30(4):1203–1205. [PubMed] [Google Scholar]
  41. Strand M. Transformation-related antigens identified by monoclonal antibodies. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3234–3238. doi: 10.1073/pnas.77.6.3234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Vassalli J. D., Hamilton J., Reich E. Macrophage plasminogen activator: induction by concanavalin A and phorbol myristate acetate. Cell. 1977 Jul;11(3):695–705. doi: 10.1016/0092-8674(77)90086-1. [DOI] [PubMed] [Google Scholar]
  43. Verma A. K., Ashendel C. L., Boutwell R. K. Inhibition by prostaglandin synthesis inhibitors of the induction of epidermal ornithine decarboxylase activity, the accumulation of prostaglandins, and tumor promotion caused by 12-O-tetradecanoylphorbol-13-acetate. Cancer Res. 1980 Feb;40(2):308–315. [PubMed] [Google Scholar]
  44. Whitfield J. F., MacManus J. P., Gillan D. J. Calcium-dependent stimulation by a phorbol ester (PMA) of thymic lymphoblast DNA synthesis and proliferation. J Cell Physiol. 1973 Oct;82(2):151–156. doi: 10.1002/jcp.1040820203. [DOI] [PubMed] [Google Scholar]
  45. Wigler M., DeFeo D., Weinstein I. B. Induction of plasminogen activator in cultured cells by macrocyclic plant diterpene esters and other agents related to tumor promotion. Cancer Res. 1978 May;38(5):1434–1437. [PubMed] [Google Scholar]
  46. Wigler M., Weinstein I. B. Tumour promotor induces plasminogen activator. Nature. 1976 Jan 22;259(5540):232–233. doi: 10.1038/259232a0. [DOI] [PubMed] [Google Scholar]
  47. Wilson E. L., Reich E. Modulation of plasminogen activator synthesis in chick embryo fibroblasts by cyclic nucleotides and phorobol myristate acetate. Cancer Res. 1979 May;39(5):1579–1586. [PubMed] [Google Scholar]
  48. Yuspa S. H., Lichti U., Ben T., Patterson E., Hennings H., Slaga T. J., Colburn N., Kelsey W. Phorbol esters stimulate DNA synthesis and ornithine decarboxylase activity in mouse epidermal cell cultures. Nature. 1976 Jul 29;262(5567):402–404. doi: 10.1038/262402a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES