Skip to main content
PMC 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 Nov;79(21):6541–6545. doi: 10.1073/pnas.79.21.6541

Retinoids and phorbol esters alter release of fibronectin from enucleated cells.

S D Bolmer, G Wolf
PMCID: PMC347163  PMID: 6959135

Abstract

Addition of 12-tetradecanoylphorbol 13-acetate (TPA) to cultures of intact Swiss mouse 3T3 fibroblasts induced a dose-dependent increase in ornithine decarboxylase (OrnDCase) activity. Over the same concentration range, 10(-9) to 10(-6) M, TPA induced the release of radioactively labeled fibronectin (FN) from the cells into the culture medium. Retinoic acid, a derivative of vitamin A, inhibited in a dose-dependent manner both the increase in OrnDCase activity and the release of FN induced by TPA. To examine the effects of TPA and retinoic acid in enucleated cells, the cells were treated with 7.5 micrograms of cytochalasin B per ml of medium during centrifugation at 10,000 X g for 35 min at 37 degrees C. In a series of five experiments, the treated cells were 94.7 +/- 4.8% (+/- SEM) enucleated as measured by [3H]thymidine incorporation and verified by Giesma staining for nuclei. In the enucleated cells, TPA did not induce increased OrnDCase activity but did induce FN release in a dose-dependent fashion over the same concentration range effective for FN release from intact cells. Moreover, addition of retinoic acid to the enucleated cells inhibited the phorbol ester-induced release of FN in a dose-dependent manner. A series of phorbol ester derivatives showed the same order of activity in causing FN release from the enucleated cells as reported for inducing OrnDcase activity in intact cells or in promoting mouse skin tumors in vivo. Similarly, several retinoids were tested for their ability to inhibit the phorbol ester-induced release of FN from enucleated cells. The efficacy of the retinoids in preventing FN release paralleled their activity in inhibiting phorbol ester-induced OrnDCase activity and skin tumor promotion, as reported in the literature. We conclude that at least one aspect of tumor promotion induced by phorbol esters--the loss of FN--does not require the cell nucleus, and further, that retinoids can inhibit this aspect of tumor promotion without nuclear involvement.

Full text

PDF

Images in this article

6544Image
on p.6544

Selected References

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

  1. Adamo S., De Luca L. M., Akalovsky I., Bhat P. V. Retinoid-induced adhesion in cultured, transformed mouse fibroblasts. J Natl Cancer Inst. 1979 Jun;62(6):1473–1478. [PubMed] [Google Scholar]
  2. Ali I. U., Hynes R. O. Effects of cytochalasin B and colchicine on attachment of a major surface protein of fibroblasts. Biochim Biophys Acta. 1977 Nov 15;471(1):16–24. doi: 10.1016/0005-2736(77)90388-1. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Colburn N. H. Tumor promoter produces anchorage independence in mouse epidermal cells by an induction mechanism. Carcinogenesis. 1980;1(11):951–954. doi: 10.1093/carcin/1.11.951. [DOI] [PubMed] [Google Scholar]
  5. De Luca L. M. The direct involvement of vitamin A in glycosyl transfer reactions of mammalian membranes. Vitam Horm. 1977;35:1–57. doi: 10.1016/s0083-6729(08)60520-8. [DOI] [PubMed] [Google Scholar]
  6. Dion L. D., De Luca L. M., Colburn N. H. Phorbol ester-induced anchorage independence and its antagonism by retinoic acid correlates with altered expression of specific glycoproteins. Carcinogenesis. 1981;2(10):951–958. doi: 10.1093/carcin/2.10.951. [DOI] [PubMed] [Google Scholar]
  7. Driedger P. E., Blumberg P. M. Quantitative correlation between in vitro and in vivo activities of phorbol esters. Cancer Res. 1979 Mar;39(3):714–719. [PubMed] [Google Scholar]
  8. 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]
  9. Fuchs E., Green H. Regulation of terminal differentiation of cultured human keratinocytes by vitamin A. Cell. 1981 Sep;25(3):617–625. doi: 10.1016/0092-8674(81)90169-0. [DOI] [PubMed] [Google Scholar]
  10. Haddox M. K., Scott K. F., Russell D. H. Retinol inhibition of ornithine decarboxylase induction and G1 progression in Chinese hamster ovary cells. Cancer Res. 1979 Dec;39(12):4930–4938. [PubMed] [Google Scholar]
  11. Hassell J. R., Pennypacker J. P., Kleinman H. K., Pratt R. M., Yamada K. M. Enhanced cellular fibronectin accumulation in chondrocytes treated with vitamin A. Cell. 1979 Aug;17(4):821–826. doi: 10.1016/0092-8674(79)90322-2. [DOI] [PubMed] [Google Scholar]
  12. Jetten A. M., Jetten M. E., Shapiro S. S., Poon J. P. Characterization of the action of retinoids on mouse fibroblast cell lines. Exp Cell Res. 1979 Mar 15;119(2):289–299. doi: 10.1016/0014-4827(79)90356-2. [DOI] [PubMed] [Google Scholar]
  13. Keski-Oja J., Shoyab M., De Larco J. E., Todaro G. J. Rapid release of fibronectin from human lung fibroblasts by biologically active phorbol esters. Int J Cancer. 1979 Aug;24(2):218–224. doi: 10.1002/ijc.2910240214. [DOI] [PubMed] [Google Scholar]
  14. Kurkinen M., Wartiovaara J., Vaheri A. Cytochalasin B releases a major surface-associated glycoprotein, fibronectin, from cultured fibroblasts. Exp Cell Res. 1978 Jan;111(1):127–137. doi: 10.1016/0014-4827(78)90243-4. [DOI] [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Lapointe D. S., Cohen R. J. A rapid and efficient microassay of ornithine decarboxylase. Anal Biochem. 1980 Dec;109(2):291–294. doi: 10.1016/0003-2697(80)90651-x. [DOI] [PubMed] [Google Scholar]
  17. Liau G., Ong D. E., Chytil F. Interaction of the retinol/cellular retinol-binding protein complex with isolated nuclei and nuclear components. J Cell Biol. 1981 Oct;91(1):63–68. doi: 10.1083/jcb.91.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lowe M. E., Pacifici M., Holtzer H. Effects of phorbol-12-myristate-13-acetate on the phenotypic program of cultured chondroblasts and fibroblasts. Cancer Res. 1978 Aug;38(8):2350–2356. [PubMed] [Google Scholar]
  19. Nagle D. S., Blumberg P. M. Activity of phorbol ester tumor promoters on enucleated Swiss 3T3 cells. Cancer Res. 1980 Apr;40(4):1066–1072. [PubMed] [Google Scholar]
  20. Newton D. L., Henderson W. R., Sporn M. B. Structure-activity relationships of retinoids in hamster tracheal organ culture. Cancer Res. 1980 Oct;40(10):3413–3425. [PubMed] [Google Scholar]
  21. O'Brien T. G., Diamond L. Ornithine decarboxylase induction and DNA synthesis in hamster embryo cell cultures treated with tumor-promoting phorbol diesters. Cancer Res. 1977 Nov;37(11):3895–3900. [PubMed] [Google Scholar]
  22. OUCHTERLONY O. Diffusion-in-gel methods for immunological analysis. Prog Allergy. 1958;5:1–78. [PubMed] [Google Scholar]
  23. Paranjpe M. S., De Larco J. E., Todaro G. J. Retinoids block ornithine decarboxylase induction in cells treated with the tumor promotor TPA or the peptide growth hormones, EGF and SGF. Biochem Biophys Res Commun. 1980 May 30;94(2):586–591. doi: 10.1016/0006-291x(80)91272-3. [DOI] [PubMed] [Google Scholar]
  24. Sasak W., De Luca L. M., Dion L. D., Silverman-Jones C. S. Effect of retinoic acid on cell surface glycopeptides of cultured spontaneously transformed mouse fibroblasts (BALB/c 3T12-3 cells). Cancer Res. 1980 Jun;40(6):1944–1949. [PubMed] [Google Scholar]
  25. Verma A. K., Shapas B. G., Rice H. M., Boutwell R. K. Correlation of the inhibition by retinoids of tumor promoter-induced mouse epidermal ornithine decarboxylase activity and of skin tumor promotion. Cancer Res. 1979 Feb;39(2 Pt 1):419–425. [PubMed] [Google Scholar]
  26. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  27. 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