DNA intercalative 4-butylaminopyrimido[4′,5′:4,5]thieno(2,3-b)quinoline induces cell cycle arrest and apoptosis in leukemia cells

Abstract

Purpose

DNA intercalators are one of the interesting groups in cancer chemotherapy. The development of novel anticancer small molecule has gained remarkable interest over the last decade. In this study, we synthesized and investigated the ability of a tetracyclic-condensed quinoline compound, 4-butylaminopyrimido[4′,5′:4,5]thieno(2,3-b)quinoline (BPTQ), to interact with double-stranded DNA and inhibit cancer cell proliferation.

Methods

Circular dichroism, topological studies, molecular docking, absorbance, and fluorescence spectral titrations were employed to study the interaction of BPTQ with DNA. Cytotoxicity was studied by performing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assay. Further, cell cycle analysis by flow cytometry, annexin V staining, mitochondrial membrane potential assay, DNA fragmentation, and western blot analysis were used to elucidate the mechanism of action of BPTQ at the cellular level.

Results

Spectral, topological, and docking studies confirmed that BPTQ is a typical intercalator of DNA. BPTQ induces dose-dependent inhibitory effect on the proliferation of cancer cells by arresting cells at S and G2/M phase. Further, BPTQ activates the mitochondria-mediated apoptosis pathway, as explicated by a decrease in mitochondrial membrane potential, increase in the Bax:Bcl-2 ratio, and activation of caspases.

Conclusion

These results confirmed that BPTQ is a DNA intercalative anticancer molecule, which could aid in the development of future cancer therapeutic agents.

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Acknowledgments

This work was supported by grant BT/PR10513/BRB/10/618/2008 to G. M. A. from Department of Biotechnology (DBT), Ministry of Science and Technology, Government of India (New Delhi). We are grateful to S. R. Inamdar for providing critical reagents; K. S. Rachana and M. S. Manu for critical reading; and P. Kondaiah, H. N. Kiran Kumar, and members of S. C. R. laboratory for help. FACS and confocal facilities of IISc, and FACS facility of C-CAMP, NCBS are also acknowledged. H. G. R was supported by DBT (India). We also thank Chethan Kumar for his assistance in docking studies.

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The authors declare no conflicts of interest.

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Correspondence to Gopal M. Advi Rao.

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RohitKumar, H.G., Asha, K.R., Raghavan, S.C. et al. DNA intercalative 4-butylaminopyrimido[4′,5′:4,5]thieno(2,3-b)quinoline induces cell cycle arrest and apoptosis in leukemia cells. Cancer Chemother Pharmacol 75, 1121–1133 (2015). https://doi.org/10.1007/s00280-015-2735-6

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Keywords

  • DNA intercalators
  • Chemotherapy
  • Apoptosis
  • Leukemia