Skip to main content
SpringerLink
Log in

Dimeric bisbenzimidazoles: Cytotoxicity and effects on DNA methylation in normal and cancer human cells

  • Cell Molecular Biology
  • Published:
Molecular Biology Aims and scope Submit manuscript

Abstract

Cancer cells are characterized by hypermethylation of the promoter regions of tumor suppressor genes. DNA methyltransferase inhibitors reactivate the genes, pointing to DNA methyltransferases as potential targets for anticancer therapy. Dimeric bisbenzimidazoles varying in the length of an oligomeric linker between two bisbenzimidazole residues (DB(n), where n is the number of methylene groups in the linker) were earlier shown to efficiently inhibit methylation of DNA duplexes by murine DNA methyltransferase Dnmt3a. Here, some of the compounds were tested for cytotoxicity, cell penetration, and effect on genomic DNA methylation in F-977 fetal lung fibroblasts and HeLa cervical cancer cells. Within the 0–60 μM concentration range, only DB(11) exerted a significant toxic effect on normal cells, whereas the effects of DB(n) on cancer cells were not significant. DB(1) and DB(3) slightly stimulated proliferation of HeLa and F-977 cells, respectively. DB(1) and DB(3) penetrated into the nuclei of HeLa and F-977 cells and accumulated predominantly in or near the nucleolus, while DB(11) was incapable of nuclear penetration. HeLa cells incubated with 26 μM DB(1) or DB(3) displayed a decrease in methylation of the 18S rRNA gene, which was in the regions of predominant accumulation of DB(1) and DB(3). The same DB(3) concentration exerted a similar effect on F-977 cells. However, the overall genomic DNA methylation level remained unchanged in both of the cell lines. The results indicated that DB(n)-type compounds can be used to demethylate certain genes and are thereby promising as potential anticancer agents.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

DB(n):

dimeric bisbenzimidazole

m5dC:

5-methyl-2′-deoxycytidine

MTase:

C5-cytosine DNA methyl-transferase

MTT:

3-[4,5-dimethylthyazol-2-yl]-2,5-diphe-nyltetrazolium bromide

DMSO:

dimethyl sulfoxide

PCR:

polymerase chain reaction

rDNA:

ribosomal repeat

References

  1. Bird A. 2002. DNA methylation patterns and epigenetic memory. Genes Dev. 16, 6–21.

    Article  PubMed  CAS  Google Scholar 

  2. Hermann A., Gowher H., Jeltsch A. 2004. Biochemistry and biology of mammalian DNA methyltransferases. Cell Mol. Life Sci. 61, 2571–2587.

    Article  PubMed  CAS  Google Scholar 

  3. Eden A., Gaudet F., Waghmare A., Jaenisch R. 2003. Chromosomal instability and tumors promoted by DNA hypomethylation. Science. 300, 455.

    Article  PubMed  CAS  Google Scholar 

  4. Mani S., Herceg Z. 2010. DNA demethylating agents and epigenetic therapy of cancer. Adv. Genetics. 70, 327–340.

    Article  CAS  Google Scholar 

  5. Kirsanova O.V., Cherepanova N.A., Gromova E.S. 2009. Inhibition of C5-cytosine-DNA methyltransferases. Biochemistry (Moscow). 74, 1175–1186.

    Article  CAS  Google Scholar 

  6. Garcia-Manero G. 2008. Demethylating agents in myeloid malignancies. Curr. Opin. Oncol. 20, 705–710.

    Article  PubMed  CAS  Google Scholar 

  7. Streltsov S.A., Gromyko A.V., Oleinikov V.A., Zhuze A.L. 2006. The Hoechst 33258 covalent dimer covers a total turn of the double-stranded DNA. J. Biomol. Struct. Dynamics. 24, 285–302.

    Article  CAS  Google Scholar 

  8. Cherepanova N.A., Ivanov A.A., Maltseva D.V., Minero A.S., Gromyko A.V., Streltsov S.A., Zhuze A.L., Gromova E.S. 2011. Dimeric bisbenzimidazoles inhibit the DNA methylation catalyzed by the murine Dnmt3a catalytic domain. J. Enzyme Inhibit. Med. Chem. 26, 295–300.

    Article  CAS  Google Scholar 

  9. Veiko N.N., Liapunova N.A., Kosyakova N.V., Spitkovskii D.M. 2001. Elements of structural organization of the transcribed area of the ribosomal repeat (rDNA) in human peripheral blood lymphocytes. Mol. Biol. (Moscow). 35, 45–55.

    Article  CAS  Google Scholar 

  10. Lyapunova N.A., Veiko N.N. 2010. Ribosomal genes in the human genome: Identification of four fractions, their organization in the nucleolus and metaphase chromosomes. Russ. J. Genet. 46, 1070–1073.

    Article  CAS  Google Scholar 

  11. Costello J.F., Fruhwald M.C., Smiraglia D.J., Rush L.J., Robertson G.P., Gao X., Wright F.A., Fera-misco J.D., Peltomaki P., Lang J.C., Schuller D.E., Yu L., Bloomfield C.D., Caligiuri M.A., Yates A., Nishikawa R., Su Huang H., Petrelli N.J., Zhang X., O’Dorisio M.S., Held W.A., Cavenee W.K., Plass C. 2000. Aberrant CpG-island methylation has non-random and tumour-type-specific patterns. Nature Genet. 24, 132–138.

    Article  PubMed  CAS  Google Scholar 

  12. Esteller M. 2007. Epigenetic gene silencing in cancer: The DNA hypermethylome. Hum. Mol. Genet. 16(1), R50–R59.

    Article  PubMed  CAS  Google Scholar 

  13. Ren J., Singh B.N., Huang Q., Li Z., Gao Y., Mishra P., Hwa Y.L., Li J., Dowdy S.C., Jiang S.W. 2011. DNA hypermethylation as a chemotherapy target. Cell Signal. 23, 1082–1093.

    Article  PubMed  CAS  Google Scholar 

  14. Fang M.Z., Wang Y., Ai N., Hou Z., Sun Y., Lu H., Welsh W., Yang C.S. 2003. Tea polyphenol (-)-epigallocatechin-3-gallate inhibits DNA methyltransferase and reactivates methylation-silenced genes in cancer cell lines. Cancer Res. 63, 7563–7570.

    PubMed  CAS  Google Scholar 

  15. Villar-Garea A., Fraga M.F., Espada J., Esteller M. 2003. Procaine is a DNA-demethylating agent with growth-inhibitory effects in human cancer cells. Cancer Res. 63, 4984–4989.

    PubMed  CAS  Google Scholar 

  16. Lee B.H., Yegnasubramanian S., Lin X., Nelson W.G. 2005. Procainamide is a specific inhibitor of DNA methyltransferase 1. J. Biol. Chem. 280, 40749–40756.

    Article  PubMed  CAS  Google Scholar 

  17. Segura-Pacheco B., Trejo-Becerril C., Perez-Cardenas E., Taja-Chayeb L., Mariscal I., Chavez A., Acuna C., Salazar A.M., Lizano M., Duenas-Gonzalez A. 2003. Reactivation of tumor suppressor genes by the cardiovascular drugs hydralazine and procainamide and their potential use in cancer therapy. Clin. Cancer Res.: Official J. Am. Assoc. Cancer Res. 9, 1596–1603.

    CAS  Google Scholar 

  18. Datta J., Ghoshal K., Denny W.A., Gamage S.A., Brooke D.G., Phiasivongsa P., Redkar S., Jacob S.T. 2009. A new class of quinoline-based DNA hypomethylating agents reactivates tumor suppressor genes by blocking DNA methyltransferase 1 activity and inducing its degradation. Cancer Res. 69, 4277–4285.

    Article  PubMed  CAS  Google Scholar 

  19. Ivanov A.A., Salyanov V.I., Streltsov S.A., Cherepanova N.A., Gromova E.S., Zhuze A.L. 2011. DNA sequence-specific ligands: XIV. Synthesis of fluorescent biologicaly active dimeric bisbenzimidazoles, DB(3, 4, 5, 7, 11). Russ. J. Bioorg. Chem. 37, 472–482.

    Google Scholar 

  20. Petrova N.V., Iarovaia O.V., Verbovoy V.A., Razin S.V. 2005. Specific radial positions of centromeres of human chromosomes X, 1, and 19 remain unchanged in chromatin-depleted nuclei of primary human fibroblasts: Evidence for the organizing role of the nuclear matrix. J. Cell. Biochemistry. 96, 850–857.

    Article  CAS  Google Scholar 

Download references

Author information

Author notes

  1. A. A. Ivanov

    Present address: Institute of Carcinogenesis, Blokhin Cancer Research Center, Russian Academy of Medical Sciences, Moscow, 115478, Russia

Authors and Affiliations

  1. Faculty of Chemistry, Moscow State University, Moscow, 119991, Russia

    M. V. Darii, A. R. Rakhimova, V. N. Tashlitsky & E. S. Gromova

  2. Medical Genetic Research Center, Russian Academy of Medical Sciences, Moscow, 115478, Russia

    S. V. Kostyuk & N. N. Veiko

  3. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991, Russia

    A. A. Ivanov & A. L. Zhuze

Authors
  1. M. V. Darii
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. R. Rakhimova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. V. N. Tashlitsky
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S. V. Kostyuk
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. N. N. Veiko
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A. A. Ivanov
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A. L. Zhuze
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. E. S. Gromova
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to E. S. Gromova.

Additional information

Original Russian Text © M.V. Darii, A.R. Rakhimova, V.N. Tashlitsky, S.V. Kostyuk, N.N. Veiko, A.A. Ivanov, A.L. Zhuze, E.S. Gromova, 2013, published in Molekulyarnaya Biologiya, 2013, Vol. 47, No. 2, pp. 292–301.

These authors contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Darii, M.V., Rakhimova, A.R., Tashlitsky, V.N. et al. Dimeric bisbenzimidazoles: Cytotoxicity and effects on DNA methylation in normal and cancer human cells. Mol Biol 47, 259–266 (2013). https://doi.org/10.1134/S0026893313020040

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0026893313020040

Keywords

Search

Navigation