Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1971 Sep 1;50(3):746–761. doi: 10.1083/jcb.50.3.746

ON THE DIFFERENTIAL CYTOTOXICITY OF ACTINOMYCIN D

Stanley G Sawicki 1, Gabriel C Godman 1
PMCID: PMC2108313  PMID: 4398631

Abstract

Actinomycin D (AMD) at concentrations that inhibit cellular RNA synthesis by 85% or more causes an acute phase of lethal cell degeneration in HeLa cultures beginning as early as 3 hr after drug exposure, resulting in the nearly complete loss of viable cells by 12 hr. The loss of cells during this acute phase of lethality is closely dose dependent. Vero, WI38, or L cells are not susceptible to this early acute cyto-intoxication by AMD, and may begin to die only after 1–2 days. Differential susceptibility to acute cyto-intoxication by AMD, or other inhibitors of RNA synthesis (daunomycin or nogalamycin), among different types of cultured cells is analogous to that observed in vivo in certain tissues and tumors, and cannot be accounted for by differences in the effect of AMD on RNA, DNA, or protein syntheses, or by the over-all loss of preformed RNA. Actinomycin D in a dose that inhibits RNA synthesis causes an equivalent loss of the prelabeled RNA in all the cell types studied. Inhibition of protein synthesis with streptovitacin A or of DNA synthesis with hydroxyurea does not cause acute lethal injury in HeLa cells as does inhibition of RNA synthesis. Furthermore, since Vero or L cells divide at about the same rate as HeLa cells, no correlation can be drawn between the rate of cell proliferation and susceptibility to the cytotoxicity of AMD. Susceptibile cells are most vulnerable to intoxication by AMD in the G1-S interphase or early S phase. Inhibition of protein synthesis (which protects cells against damage by other agents affecting DNA) does not protect against AMD-induced injury. Although HeLa cells bind more AMD at a given dose than Vero or L cells, the latter cell types, given higher doses, can be made to bind proportionally more AMD without succumbing to acute cyto-intoxication. It is suggested that the differential susceptibility of these cell types to acute poisoning by AMD may reflect differences among various cells in the function or stability of certain RNA species not directly involved in translation whose presence is vital to cells. In HeLa cells, these critical species of RNA are presumed to have a short half-life.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Attardi G., Parnas H., Hwang M. I., Attardi B. Giant-size rapidly labeled nuclear ribonucleic acid and cytoplasmic messenger ribonucleic acid in immature duck erythrocytes. J Mol Biol. 1966 Sep;20(1):145–182. doi: 10.1016/0022-2836(66)90123-9. [DOI] [PubMed] [Google Scholar]
  2. Bacchetti S., Whitmore G. F. Actinomycin D: effects on mouse L-cells. Biophys J. 1969 Dec;9(12):1427–1445. doi: 10.1016/S0006-3495(69)86463-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Darzynkiewicz Z., Bolund L., Ringertz N. R. Actinomycin binding of normal and phytohaemagglutinin stimulated lymphocytes. Exp Cell Res. 1969 Apr;55(1):120–122. doi: 10.1016/0014-4827(69)90465-0. [DOI] [PubMed] [Google Scholar]
  4. Darzynkiewicz Z., Gledhill B. L., Ringertz N. R. Changes in deoxyribonucleo-protein during spermiogenesis in the bull. 3 H-Actinomycin D binding capacity. Exp Cell Res. 1969 Dec;58(2):435–438. doi: 10.1016/0014-4827(69)90527-8. [DOI] [PubMed] [Google Scholar]
  5. Dingman C. W., Peacock A. C. Analytical studies on nuclear ribonucleic acid using polyacrylamide gel electrophoresis. Biochemistry. 1968 Feb;7(2):659–668. doi: 10.1021/bi00842a022. [DOI] [PubMed] [Google Scholar]
  6. Djordjevic B., Kim J. H. Different lethal effects of mitomycin C and actinomycin D during the division cycle of HeLa cells. J Cell Biol. 1968 Sep;38(3):477–482. doi: 10.1083/jcb.38.3.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Elkind M. M., Kano E., Sutton-Gilbert H. Cell killing by actinomycin D in relation to the growth cycle of chinese hamster cells. J Cell Biol. 1969 Aug;42(2):366–377. doi: 10.1083/jcb.42.2.366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Farber E., Baserga R. Differential effects of hydroxyurea on survival of proliferating cells in vivo. Cancer Res. 1969 Jan;29(1):136–139. [PubMed] [Google Scholar]
  9. GEORGIEV G. P., SAMARINA O. P., LERMAN M. I., SMIRNOV M. N., SEVERTZOV A. N. BIOSYNTHESIS OF MESSENGER AND RIBOSOMAL RIBONUCLEIC ACIDS IN THE NUCLEOLOCHROMOSOMAL APPARATUS OF ANIMAL CELLS. Nature. 1963 Dec 28;200:1291–1294. doi: 10.1038/2001291a0. [DOI] [PubMed] [Google Scholar]
  10. Goldstein R., Okada S. Interphase death of cultured mammalian cells (L5178Y). Radiat Res. 1969 Aug;39(2):361–373. [PubMed] [Google Scholar]
  11. HARRIS H. FUNCTION OF THE SHORT-LIVED RIBONUCLEIC ACID IN THE CELL NUCLEUS. Nature. 1964 Feb 29;201:863–867. doi: 10.1038/201863a0. [DOI] [PubMed] [Google Scholar]
  12. Harris H. The reactivation of the red cell nucleus. J Cell Sci. 1967 Mar;2(1):23–32. doi: 10.1242/jcs.2.1.23. [DOI] [PubMed] [Google Scholar]
  13. Houssais J. F., Attardi G. High molecular weight nonribosomal-type nuclear RNA and cytoplasmic messenger RNA in HeLa cells. Proc Natl Acad Sci U S A. 1966 Aug;56(2):616–623. doi: 10.1073/pnas.56.2.616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hyman R. W., Davidson N. Kinetics of the in vitro inhibition of transcription by actinomycin. J Mol Biol. 1970 Jun 14;50(2):421–438. doi: 10.1016/0022-2836(70)90202-0. [DOI] [PubMed] [Google Scholar]
  15. JHEE H. T., HAN S. S. EFFECTS OF ACTINOMYCIN D ON THE SALIVARY GLANDS OF THE RAT. Life Sci. 1964 Nov;3:1239–1247. doi: 10.1016/0024-3205(64)90042-6. [DOI] [PubMed] [Google Scholar]
  16. Kim J. H., Perez A. G., Djordjevic B. Studies on unbalanced growth in synchronized HeLa cells. Cancer Res. 1968 Dec;28(12):2443–2447. [PubMed] [Google Scholar]
  17. Lorch I. J., Jeon K. W. Reversible effect of actinomycin D on nucleoli of Amoeba discoides: nuclear transplantation study. Nature. 1969 Mar 15;221(5185):1073–1074. doi: 10.1038/2211073a0. [DOI] [PubMed] [Google Scholar]
  18. Magee W. E., Miller O. V. Initiation of vaccinia virus infection in actinomycin D-pretreated cells. J Virol. 1968 Jul;2(7):678–685. doi: 10.1128/jvi.2.7.678-685.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mauro F., Madoc-Jones H. Age responses of cultured mammalian cells to cytotoxic drugs. Cancer Res. 1970 May;30(5):1397–1408. [PubMed] [Google Scholar]
  20. Nakamura T., Prestayko A. W., Busch H. Studies on nucleolar 4 to 6 S ribonucleic acid of Novikoff hepatoma cells. J Biol Chem. 1968 Apr 10;243(7):1368–1375. [PubMed] [Google Scholar]
  21. Penman S., Vesco C., Penman M. Localization and kinetics of formation of nuclear heterodisperse RNA, cytoplasmic heterodisperse RNA and polyribosome-associated messenger RNA in HeLa cells. J Mol Biol. 1968 May 28;34(1):49–60. doi: 10.1016/0022-2836(68)90234-9. [DOI] [PubMed] [Google Scholar]
  22. Perry R. P. THE CELLULAR SITES OF SYNTHESIS OF RIBOSOMAL AND 4S RNA. Proc Natl Acad Sci U S A. 1962 Dec;48(12):2179–2186. doi: 10.1073/pnas.48.12.2179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pfeiffer S. E., Tolmach L. J. Inhibition of DNA synthesis in HeLa cells by hydroxyurea. Cancer Res. 1967 Jan;27(1):124–129. [PubMed] [Google Scholar]
  24. Philips F. S., Sternberg S. S., Schwartz H. S., Cronin A. P., Sodergren J. E., Vidal P. M. Hydroxyurea. I. Acute cell death in proliferating tissues in rats. Cancer Res. 1967 Jan;27(1):61–75. [PubMed] [Google Scholar]
  25. Phillips R. A., Tolmach L. J. Repair of potentially lethal damage in x-irradiated HeLa cells. Radiat Res. 1966 Nov;29(3):413–432. [PubMed] [Google Scholar]
  26. Prescott D. M., Goldstein L. Nuclear-cytoplasmic interaction in DNA synthesis. Science. 1967 Jan 27;155(3761):469–470. doi: 10.1126/science.155.3761.469. [DOI] [PubMed] [Google Scholar]
  27. Prestayko A. W., Tonato M., Busch H. Low molecular weight RNA associated with 28 s nucleolar RNA. J Mol Biol. 1970 Feb 14;47(3):505–515. doi: 10.1016/0022-2836(70)90318-9. [DOI] [PubMed] [Google Scholar]
  28. Reich E., Goldberg I. H. Actinomycin and nucleic acid function. Prog Nucleic Acid Res Mol Biol. 1964;3:183–234. doi: 10.1016/s0079-6603(08)60742-4. [DOI] [PubMed] [Google Scholar]
  29. Roberts W. K., Newman J. F. Use of low concentrations of actinomycin D in the study of RNA synthesis in Ehrlich ascites cells. J Mol Biol. 1966 Sep;20(1):63–73. doi: 10.1016/0022-2836(66)90117-3. [DOI] [PubMed] [Google Scholar]
  30. Rovera G., Berman S., Baserga R. Pulse labeling of RNA of mammalian cells. Proc Natl Acad Sci U S A. 1970 Apr;65(4):876–883. doi: 10.1073/pnas.65.4.876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. SCHERRER K., LATHAM H., DARNELL J. E. Demonstration of an unstable RNA and of a precursor to ribosomal RNA in HeLa cells. Proc Natl Acad Sci U S A. 1963 Feb 15;49:240–248. doi: 10.1073/pnas.49.2.240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Scherrer K., Marcaud L., Zajdela F., London I. M., Gros F. Patterns of RNA metabolism in a differentiated cell: a rapidly labeled, unstable 60S RNA with messenger properties in duck erythroblasts. Proc Natl Acad Sci U S A. 1966 Nov;56(5):1571–1578. doi: 10.1073/pnas.56.5.1571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schwartz H. S., Sodergren J. E. Actinomycin D effects on nucleic acids during tumor regression. Cancer Res. 1968 Mar;28(3):445–451. [PubMed] [Google Scholar]
  34. Schwartz H. S., Sodergren J. E., Ambaye R. Y. Actinomycin D: drug concentrations and actions in mouse tissues and tumors. Cancer Res. 1968 Feb;28(2):192–197. [PubMed] [Google Scholar]
  35. Shearer R. W., McCarthy B. J. Evidence for ribonucleic acid molecules restricted to the cell nucleus. Biochemistry. 1967 Jan;6(1):283–289. doi: 10.1021/bi00853a044. [DOI] [PubMed] [Google Scholar]
  36. Shipman C., Jr Evaluation of 4-(2-hydroxyethyl)-1-piperazineëthanesulfonic acid (HEPES) as a tissue culture buffer. Proc Soc Exp Biol Med. 1969 Jan;130(1):305–310. doi: 10.3181/00379727-130-33543. [DOI] [PubMed] [Google Scholar]
  37. Sinclair W. K. Hydroxyurea: differential lethal effects on cultured mammalian cells during the cell cycle. Science. 1965 Dec 24;150(3704):1729–1731. doi: 10.1126/science.150.3704.1729. [DOI] [PubMed] [Google Scholar]
  38. Soeiro R., Birnboim H. C., Darnell J. E. Rapidly labeled HeLa cell nuclear RNA. II. Base composition and cellular localization of a heterogeneous RNA fraction. J Mol Biol. 1966 Aug;19(2):362–372. doi: 10.1016/s0022-2836(66)80010-4. [DOI] [PubMed] [Google Scholar]
  39. Soeiro R., Vaughan M. H., Warner J. R., Darnell J. E., Jr The turnover of nuclear DNA-like RNA in HeLa cells. J Cell Biol. 1968 Oct;39(1):112–118. doi: 10.1083/jcb.39.1.112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Stewart G. A., Farber E. The rapid acceleration of hepatic nuclear ribonucleic acid breakdown by actinomycin but not by ethionine. J Biol Chem. 1968 Sep 10;243(17):4479–4485. [PubMed] [Google Scholar]
  41. Thompson L. R., McCarthy B. J. Stimulation of nuclear DNA and RNA synthesis by cytoplasmic extracts in vitro. Biochem Biophys Res Commun. 1968 Jan 25;30(2):166–172. doi: 10.1016/0006-291x(68)90465-8. [DOI] [PubMed] [Google Scholar]
  42. Tomkins G. M., Gelehrter T. D., Granner D., Martin D., Jr, Samuels H. H., Thompson E. B. Control of specific gene expression in higher organisms. Expression of mammalian genes may be controlled by repressors acting on the translation of messenger RNA. Science. 1969 Dec 19;166(3912):1474–1480. doi: 10.1126/science.166.3912.1474. [DOI] [PubMed] [Google Scholar]
  43. Warner J. R., Soeiro R., Birnboim H. C., Girard M., Darnell J. E. Rapidly labeled HeLa cell nuclear RNA. I. Identification by zone sedimentation of a heterogeneous fraction separate from ribosomal precursor RNA. J Mol Biol. 1966 Aug;19(2):349–361. doi: 10.1016/s0022-2836(66)80009-8. [DOI] [PubMed] [Google Scholar]
  44. Weinberg R. A., Penman S. Small molecular weight monodisperse nuclear RNA. J Mol Biol. 1968 Dec;38(3):289–304. doi: 10.1016/0022-2836(68)90387-2. [DOI] [PubMed] [Google Scholar]
  45. Zischka R., Langlois A. J., Bonar R. A., Rao P. R., Beard J. W. Effects of actinomycin D on avian myeloblast and BAI strain A virus RNA synthesis in vitro. Cancer Res. 1966 Sep;26(9):1839–1846. [PubMed] [Google Scholar]

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

RESOURCES