Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1974 Jan 1;60(1):249–257. doi: 10.1083/jcb.60.1.249

INHIBITION OF CELL GROWTH IN THE G1 PHASE BY ADENOSINE 3',5'-CYCLIC MONOPHOSPHATE

Jeffrey E Froehlich 1, Martin Rachmeler 1
PMCID: PMC2109140  PMID: 4358429

Abstract

Incorporation of tritiated thymidine into acid-precipitable material was used to measure the rate of DNA synthesis in secondary cultures of human diploid fibroblasts. Confluent cultures of human diploid fibroblasts, which are synchronized in the G1 phase due to contact inhibition, were released from growth inhibition either by the addition of fresh medium to the cultures or by trypsinization and replating at nonconfluent densities. Either treatment resulted in a synchronous wave of DNA synthesis beginning 10–15 h after treatment and peaking at 20–25 h. In confluent cultures stimulated by fresh medium, either the addition of 0.25 mM N6, O2-dibutyryl-adenosine 3',5'-cyclic monophosphate (db-cAMP) to the medium in the interval 4–8 h after stimulation or the replacement of the fresh medium in that same 4 h interval with the depleted medium present on the cells for the 2 day period before stimulation delayed the synchronous onset of DNA synthesis in the cultures by about 4 h. In nonconfluent cultures freshly seeded from trypsinized confluent cultures, this same depleted medium obtained after a 2 day incubation of fresh medium on confluent cultures is shown to support the progress of the cells into S phase; however, the addition of 0.25 mM db-cAMP to the medium 3½ h after replating still partially prevented the initiation of DNA synthesis in the cultures. The results are discussed in terms of the role of serum and cAMP in the control of cell growth in fibroblast cultures.

Full Text

The Full Text of this article is available as a PDF (661.1 KB).

Selected References

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

  1. Borek C., Grob M., Burger M. M. Surface alterations in transformed epithelial and fibroblastic cells in culture: a disturbance of membrane degradation versus biosynthesis? Exp Cell Res. 1973 Mar 15;77(1):207–215. doi: 10.1016/0014-4827(73)90569-7. [DOI] [PubMed] [Google Scholar]
  2. Buck C. A., Glick M. C., Warren L. A comparative study of glycoproteins from the surface of control and Rous sarcoma virus transformed hamster cells. Biochemistry. 1970 Nov 10;9(23):4567–4576. doi: 10.1021/bi00825a016. [DOI] [PubMed] [Google Scholar]
  3. Buck C. A., Glick M. C., Warren L. Effect of growth on the glycoproteins from the surface of control and Rous sarcoma virus transformed hamster cells. Biochemistry. 1971 May 25;10(11):2176–2180. doi: 10.1021/bi00787a034. [DOI] [PubMed] [Google Scholar]
  4. Burger M. M., Bombik B. M., Breckenridge B. M., Sheppard J. R. Growth control and cyclic alterations of cyclic AMP in the cell cycle. Nat New Biol. 1972 Oct 11;239(93):161–163. doi: 10.1038/newbio239161a0. [DOI] [PubMed] [Google Scholar]
  5. Burger M. M. Proteolytic enzymes initiating cell division and escape from contact inhibition of growth. Nature. 1970 Jul 11;227(5254):170–171. doi: 10.1038/227170a0. [DOI] [PubMed] [Google Scholar]
  6. Bürk R. R. One-step growth cycle for BHK21-13 hamster fibroblasts. Exp Cell Res. 1970 Dec;63(2):309–316. doi: 10.1016/0014-4827(70)90218-1. [DOI] [PubMed] [Google Scholar]
  7. Ceccarini C., Eagle H. Induction and reversal of contact inhibition of growth by pH modification. Nat New Biol. 1971 Oct 27;233(43):271–273. doi: 10.1038/newbio233271a0. [DOI] [PubMed] [Google Scholar]
  8. Ceccarini C., Eagle H. pH as a determinant of cellular growth and contact inhibition. Proc Natl Acad Sci U S A. 1971 Jan;68(1):229–233. doi: 10.1073/pnas.68.1.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cox R. P., Gesner B. M. Studies on the effects of simple sugars on mammalian cells in culture and characterization of the inhibition of 3T3 fibroblasts by L-fucose. Cancer Res. 1968 Jun;28(6):1162–1172. [PubMed] [Google Scholar]
  10. Cumar F. A., Brady R. O., Kolodny E. H., McFarland V. W., Mora P. T. Enzymatic block in the synthesis of gangliosides in DNA virus-transformed tumorigenic mouse cell lines. Proc Natl Acad Sci U S A. 1970 Oct;67(2):757–764. doi: 10.1073/pnas.67.2.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  12. Heidrick M. L., Ryan W. L. Cyclic nucleotides on cell growth in vitro. Cancer Res. 1970 Feb;30(2):376–378. [PubMed] [Google Scholar]
  13. Nilausen K., Green H. Reversible arrest of growth in G1 of an established fibroblast line (3T3). Exp Cell Res. 1965 Oct;40(1):166–168. doi: 10.1016/0014-4827(65)90306-x. [DOI] [PubMed] [Google Scholar]
  14. Onodera K., Sheinin R. Macromolecular glucosamine-containing component of the surface of cultivated mouse cells. J Cell Sci. 1970 Sep;7(2):337–355. doi: 10.1242/jcs.7.2.337. [DOI] [PubMed] [Google Scholar]
  15. Otten J., Johnson G. S., Pastan I. Cyclic AMP levels in fibroblasts: relationship to growth rate and contact inhibition of growth. Biochem Biophys Res Commun. 1971 Sep;44(5):1192–1198. doi: 10.1016/s0006-291x(71)80212-7. [DOI] [PubMed] [Google Scholar]
  16. Otten J., Johnson G. S., Pastan I. Regulation of cell growth by cyclic adenosine 3',5'-monophosphate. Effect of cell density and agents which alter cell growth on cyclic adenosine 3',5'-monophosphate levels in fibroblasts. J Biol Chem. 1972 Nov 10;247(21):7082–7087. [PubMed] [Google Scholar]
  17. PUCK T. T., CIECIURA S. J., FISHER H. W. Clonal growth in vitro of human cells with fibroblastic morphology; comparison of growth and genetic characteristics of single epithelioid and fibroblast-like cells from a variety of human organs. J Exp Med. 1957 Jul 1;106(1):145–158. doi: 10.1084/jem.106.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Roth S., McGuire E. J., Roseman S. An assay for intercellular adhesive specificity. J Cell Biol. 1971 Nov;51(21):525–535. doi: 10.1083/jcb.51.2.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Roth S., McGuire E. J., Roseman S. Evidence for cell-surface glycosyltransferases. Their potential role in cellular recognition. J Cell Biol. 1971 Nov;51(21):536–547. doi: 10.1083/jcb.51.2.536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rozengurt E., Pardee A. B. Opposite effects of dibutyryl adenosine 3':5' cyclic monophosphate and serum on growth of Chinese hamster cells. J Cell Physiol. 1972 Oct;80(2):273–279. doi: 10.1002/jcp.1040800215. [DOI] [PubMed] [Google Scholar]
  21. Ryan W. L., Heidrick M. L. Inhibition of cell growth in vitro by adenosine 3',5'-monophosphate. Science. 1968 Dec 27;162(3861):1484–1485. doi: 10.1126/science.162.3861.1484. [DOI] [PubMed] [Google Scholar]
  22. SIEKEVITZ P. Uptake of radioactive alanine in vitro into the proteins of rat liver fractions. J Biol Chem. 1952 Apr;195(2):549–565. [PubMed] [Google Scholar]
  23. Sefton B. M., Rubin H. Release from density dependent growth inhibition by proteolytic enzymes. Nature. 1970 Aug 22;227(5260):843–845. doi: 10.1038/227843a0. [DOI] [PubMed] [Google Scholar]
  24. Sheppard J. R. Difference in the cyclic adenosine 3',5'-monophosphate levels in normal and transformed cells. Nat New Biol. 1972 Mar 1;236(61):14–16. doi: 10.1038/newbio236014a0. [DOI] [PubMed] [Google Scholar]
  25. Sheppard J. R. Restoration of contact-inhibited growth to transformed cells by dibutyryl adenosine 3':5'-cyclic monophosphate. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1316–1320. doi: 10.1073/pnas.68.6.1316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Temin H. M. Stimulation by serum of multiplication of stationary chicken cells. J Cell Physiol. 1971 Oct;78(2):161–170. doi: 10.1002/jcp.1040780202. [DOI] [PubMed] [Google Scholar]
  27. Wu H. C., Meezan E., Black P. H., Robbins P. W. Comparative studies on the carbohydrate-containing membrane components of normal and virus-transformed mouse fibroblasts. I. Glucosamine-labeling patterns in 3T3, spontaneously transformed 3T3, and SV-40-transformed 3T3 cells. Biochemistry. 1969 Jun;8(6):2509–2517. doi: 10.1021/bi00834a038. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES