Skip to main content
NCBI home page
Cytotechnology logoLink to Cytotechnology
. 1998 Nov;28(1-3):65–72. doi: 10.1023/A:1008017328061

Modulation of cell cycle progression and of antibody production in mouse hybridomas by a nucleotide analogue

František Franěk 1,✉, Antonín Holý 1, Ivan Votruba 1, Tomáš Eckschlager 2
PMCID: PMC3449840  PMID: 19003408

Abstract

The nucleotide analogue 9-[2-(phosphonomethoxy)ethyl]guanine (PMEG) has been identified as a powerful antiproliferative substance when acting on hybridoma cells. In the range of 10 nM to 100 nM concentrations this agent reduces cell growth rate, while its apoptosis-inducing activity is marginal. Marked induction of apoptosis can be observed at micromolar and higher order concentrations. In PMEG-supplemented media the cell cycle progression is perturbed, the flow-cytometric DNA profile shows a higher proportion of cells in the S and G2/M phases of the cell cycle. Concomitantly with the reduction of the growth rate, the specific monoclonal antibody production rate may rise by 20–27%. Addition of PMEG at the end of the exponential phase of a batch culture results in an enhancement of the final monoclonal antibody concentration.

Keywords: acyclic nucleoside phosphonate, cell cycle, hybridoma, specific MAb production rate

Full Text

The Full Text of this article is available as a PDF (97.1 KB).

References

  1. Fenge C, Fraune E, Freitag R, Scheper T, Schügerl K. On-line monitoring of monoclonal antibody formation in high-density perfusion culture using FIA. Cytotechnology. 1991;6:55–63. doi: 10.1007/BF00353703. [DOI] [PubMed] [Google Scholar]
  2. Franěk F. Starvation-induced programmed death of hybridoma cells: Prevention by amino acid mixtures. Biotech. Bioeng. 1995;45:86–90. doi: 10.1002/bit.260450112. [DOI] [PubMed] [Google Scholar]
  3. Franěk F, Šrámková K. Cell suicide in starving hybridoma culture: survival-signal effect of some amino acids. Cytotechnology. 1996;21:81–89. doi: 10.1007/BF00364839. [DOI] [PubMed] [Google Scholar]
  4. Franěk F, Šrámková K, et al. Unbalanced media for hybridoma cell culture - An alternative reality. In: Carrondo MJT, et al., editors. Animal Cell Technology. Dordrecht: Kluwer Academic Publishers; 1997. pp. 675–680. [Google Scholar]
  5. Franěk F, Šrámková K, et al. Increasing monoclonal antibody productivity by semicontinuous substitution of production medium for growth medium. In: Merten O-W, et al., editors. Animal Cell Technology. Dordrecht: Kluwer Academic Publishers; 1998. pp. 55–57. [Google Scholar]
  6. Franěk F, Vomastek T, Dolníková J. Fragmented DNA and apoptotic bodies document the programmed way of cell death in hybridoma cultures. Cytotechnology. 1992;9:117–123. doi: 10.1007/BF02521738. [DOI] [PubMed] [Google Scholar]
  7. Fussenegger M, Mazur X, Bailey JE. A novel cytostatic process enhances the productivity of Chinese hamster ovary cells. Biotech Bioeng. 1997;55:927–939. doi: 10.1002/(SICI)1097-0290(19970920)55:6<927::AID-BIT10>3.0.CO;2-4. [DOI] [PubMed] [Google Scholar]
  8. Holý A. Isopolar phosphorus-modified nucleotide analogues. In: De Clercq E, editor. Advances in Antiviral Drug Design. Greenwich, CT (U.S.A.): JAI Press, Inc.; 1994. pp. 179–232. [Google Scholar]
  9. Holý A, Rosenberg I, Dvořáková H. Synthesis of N-(2-phosphonylmethoxyethyl)derivatives of heterocyclic bases. Collect Czech Chem Commun. 1989;54:2190–2210. doi: 10.1135/cccc19892190. [DOI] [Google Scholar]
  10. Kirchhoff S, Kröger A, Cruz H, Tümmler M, Schaper F, Köster M, Hauser H. Regulation of cell growth by IRF-1 in BHK-21 cells. Cytotechnology. 1996;22:147–156. doi: 10.1007/BF00353934. [DOI] [PubMed] [Google Scholar]
  11. Kramata P, Votruba I, Otová B, Holý A. Different inhibitory potencies of acyclic phosphonomethoxyalkyl nucleotide analogs toward DNA polymerases α, δ and ε. Mol Pharmacol. 1996;49:1005–1011. [PubMed] [Google Scholar]
  12. Kruh J. Effects of sodium butyrate, a new pharmacological agent, on cells in culture. Mol Cell Biochem. 1982;42:65–82. doi: 10.1007/BF00222695. [DOI] [PubMed] [Google Scholar]
  13. Kung AL, Zetterberg A, Sherwood SW, Schimke RT. Cytotoxic effects of cell cycle phase specific agents: Result of cell cycle perturbation. Cancer Res. 1990;50:7307–7317. [PubMed] [Google Scholar]
  14. Orren DK, Petersen LN, Bohr VA. Persistent DNA damage inhibits S-phase and G2 progression, and results in apoptosis. Mol Biol Cell. 1997;8:1129–1142. doi: 10.1091/mbc.8.6.1129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pizzorno G, Cheng YC, Handshumacher RE. Pyrimidine and purine antimetabolites. In: Holland JF, Bast RC Jr, Morton DL, Frei E III, Kufe DW, Weichselbaum RR, editors. Cancer medicine. Baltimore: Williams and Wilkins; 1997. pp. 923–948. [Google Scholar]
  16. Pisarev VM, Lee SH, Connely MC, Fridland A. Intracellular metabolism and action of acyclic nucleoside phosphonates on DNA replication. Mol Pharmacol. 1997;52:63–68. doi: 10.1124/mol.52.1.63. [DOI] [PubMed] [Google Scholar]
  17. Rose WC, Crosswell AR, Bronson JJ, Martin JC. In vivoantitumor activity of 9-[(2-phosphonylmethoxy)ethyl]-guanine and related phosphonate nucleotide analogues. J Natl Cancer Inst. 1990;82:510–512. doi: 10.1093/jnci/82.6.510. [DOI] [PubMed] [Google Scholar]
  18. Štětina R, Votruba I, Holý A, Merta A. The effect of purine phosphonomethoxyalkyl derivatives on DNA synthesis in CHO Chinese hamster cells. Neoplasma. 1993;40:373–378. [PubMed] [Google Scholar]
  19. Suzuki E, Takahashi K, Ollis DF, et al. A simple structured model predicted positively-, negatively-, or non-growth associated antibody production rate depending on culture conditions. In: Murakami H, et al., editors. Animal Cell Technology: Basic and Applied Aspects. Dordrecht: Kluwer Academic Publishers; 1992. pp. 375–381. [Google Scholar]
  20. Terada S, Fukuoka K, Fujita T, Komatsu T, Takayama S., Reed JC, Suzuki E. Anti-apoptotic genes, bag-1 and bcl-2, enabled hybridoma cells to survive under treatment for arresting cell cycle. Cytotechnology. 1997;25:17–23. doi: 10.1023/A:1007954103572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Takahashi K, Terada S, Ueda H., Makishima F, Suzuki E. Growth rate suppression of cultured animal cells enhances protein productivity. Cytotechnology. 1994;15:54–64. doi: 10.1007/BF00762379. [DOI] [PubMed] [Google Scholar]
  22. Veselý J, Merta A, Votruba I, Holý A, Rosenberg I. The cytostatic effects and mechanism of action of antiviral acyclic nucleotide analogues in L1210mouse leukemia cells. Neoplasma. 1990;37:105–110. [PubMed] [Google Scholar]

Articles from Cytotechnology are provided here courtesy of Springer Science+Business Media B.V.

RESOURCES