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.
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.
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.
These results confirmed that BPTQ is a DNA intercalative anticancer molecule, which could aid in the development of future cancer therapeutic agents.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Sadikovic B, Al-Romaih K, Squire JA, Zielenska M (2008) Cause and consequences of genetic and epigenetic alterations in human cancer. Curr Genomics 9:394–408
Robison K (2010) Application of second-generation sequencing to cancer genomics. Brief Bioinform 11:524–534
Shahabuddin MS, Nambiar M, Choudhary B, Advirao GM, Raghavan SC (2010) A novel DNA intercalator, butylamino-pyrimido[4′,5′:4,5]selenolo(2,3-b)quinoline, induces cell cycle arrest and apoptosis in leukemic cells. Invest New Drugs 28:35–48
Kastan MB, Bartek J (2004) Cell-cycle checkpoints and cancer. Nature 432:316–323
Hurley LH (2002) DNA and its associated processes as targets for cancer therapy. Nat Rev Cancer 2:188–200
Martínez R, Chacón-García L (2005) The search of DNA-intercalators as antitumoral drugs: what it worked and what did not work. Curr Med Chem 12:127–151
Thurston DE (1999) Nucleic acid targeting: therapeutic strategies for the 21st century. Br J Cancer 80:65–85
Li S, Cooper VR, Thonhauser T, Lundqvist BI, Langreth DC (2009) Stacking interactions and DNA intercalation. J Phys Chem B 113:11166–11172
Nitiss JL (2009) Targeting DNA topoisomerase II in cancer chemotherapy. Nat Rev Cancer 9:338–350
Park E-J, Kwon H-K, Choi Y-M, Shin H-J, Choi S (2012) Doxorubicin induces cytotoxicity through upregulation of perk-dependent ATF3. PLoS One 7:e44990
Khan SN, Lal SK, Kumar P, Khan AU (2010) Effect of mitoxantrone on proliferation dynamics and cell-cycle progression. Biosci Rep 30:375–381
Kuo P-L, Hsu Y-L, Chang C-H, Lin C-C (2005) The mechanism of ellipticine-induced apoptosis and cell cycle arrest in human breast MCF-7 cancer cells. Cancer Lett 223:293–301
Lafayette EA, Vitalino de Almeida SM, Pitta MGDR, Carneiro Beltrão EI, da Silva TG, Olímpio de Moura R, Pitta IDR, de Carvalho LB, de Lima MDCA (2013) Synthesis, DNA binding and topoisomerase I inhibition activity of thiazacridine and imidazacridine derivatives. Molecules 18:15035–15050
Seo J, Lee HS, Lee M, Kim M, Shin C-G (2004) DA-125, a new antitumor agent, inhibits topoisomerase II as topoisomerase poison and DNA intercalator simultaneously. Arch Pharm Res 27:77–82
Prajapati SM, Patel KD, Vekariya RH, Panchal SN, Patel HD (2014) Recent advances in the synthesis of quinolines: a review. RSC Adv 4:24463–24476
Solomon VR, Lee H (2011) Quinoline as a privileged scaffold in cancer drug discovery. Curr Med Chem 18:1488–1508
Shenoy S, Vasania VS, Gopal M, Mehta A (2007) 8-Methyl-4-(3-diethylaminopropylamino) pyrimido [4′,5′;4,5] thieno (2,3-b) quinoline (MDPTQ), a quinoline derivate that causes ROS-mediated apoptosis in leukemia cell lines. Toxicol Appl Pharmacol 222:80–88
Shahabuddin MS, Gopal M, Raghavan SC (2009) Intercalating, cytotoxic, antitumour activity of 8-chloro and 4-morpholinopyrimido [4′,5′:4,5]thieno(2,3-b)quinolines. J Photochem Photobiol B 94:13–19
Gopal M, Shenoy S, Doddamani L (2003) Antitumor activity of 4-amino and 8-methyl-4-(3diethylamino propylamino)pyrimido[4′,5′:4,5]thieno (2,3-b) quinolines. J Photochem Photobiol B Biol 72:69–78
Ren J, Chaires JB (1999) Sequence and structural selectivity of nucleic acid binding ligands. Biochemistry 38:16067–16075
Raj TT, Ambekar SY (1988) Synthesis of 4-amino pyrimido [4′,5′:4,5] thieno (2,3-b) quinolines. J Chem Eng Data 33:530–531
Benesi HA, Hildebrand JH (1949) A spectrophotometric investigation of the interaction of iodine with aromatic hydrocarbons. J Am Chem Soc 71:2703–2707
Huo D, Yang L, Hou C, Fa H, Luo X, Lu Y, Zheng X, Yang J, Yang L (2009) Molecular interactions of monosulfonate tetraphenylporphyrin (TPPS1) and meso-tetra(4-sulfonatophenyl)porphyrin (TPPS) with dimethyl methylphosphonate (DMMP). Spectrochim Acta A Mol Biomol Spectrosc 74:336–343
Cao Y, He XW (1998) Studies of interaction between safranine T and double helix DNA by spectral methods. Spectrochim Acta A Mol Biomol Spectrosc 54A:883–892
Tu LC, Chen C-S, Hsiao I-C, Chern J-W, Lin C-H, Shen Y-C, Yeh SF (2005) The beta-carboline analog Mana-Hox causes mitotic aberration by interacting with DNA. Chem Biol 12:1317–1324
Canals A, Purciolas M, Aymamí J, Coll M (2005) The anticancer agent ellipticine unwinds DNA by intercalative binding in an orientation parallel to base pairs. Acta Crystallogr D Biol Crystallogr 61:1009–1012
Ritchie DW, Kemp GJ (2000) Protein docking using spherical polar Fourier correlations. Proteins 39:178–194
Hegde M, Karki SS, Thomas E, Kumar S, Panjamurthy K, Ranganatha SR, Rangappa KS, Choudhary B, Raghavan SC (2012) Novel levamisole derivative induces extrinsic pathway of apoptosis in cancer cells and inhibits tumor progression in mice. PLoS One 7:e43632
Kavitha CV, Nambiar M, Ananda Kumar CS, Choudhary B, Muniyappa K, Rangappa KS, Raghavan SC (2009) Novel derivatives of spirohydantoin induce growth inhibition followed by apoptosis in leukemia cells. Biochem Pharmacol 77:348–363
Zhou N, Xiao H, Li T-K, Nur-E-Kamal A, Liu LF (2003) DNA damage-mediated apoptosis induced by selenium compounds. J Biol Chem 278:29532–29537
Srivastava M, Nambiar M, Sharma S, Karki SS, Goldsmith G, Hegde M, Kumar S, Pandey M, Singh RK, Ray P, Natarajan R, Kelkar M, De A, Choudhary B, Raghavan SC (2012) An inhibitor of nonhomologous end-joining abrogates double-strand break repair and impedes cancer progression. Cell 151:1474–1487
Zhou C, Wu Y, Yang P (2010) Synthesis, characterization, and studies on DNA binding of the complex Fe(Sal2dienNO3, H2O). Biochem (Mosc) 75:505–512
Garrett RH, Grisham CM (2010) Biochemistry. Cengage Learning, Boston
Piantanida I, Palm BS, Zinić M, Schneider H-J (2001) A new 4,9-diazapyrenium intercalator for single- and double-stranded nucleic acids: distinct differences from related diazapyrenium compounds and ethidium bromide. J Chem Soc Perkin Trans 2:1808–1816
Amutha R, Subramanian V, Nair BU (2001) Interaction of benzidine with DNA: experimental and modelling studies. Chem Phys Lett 344:40–48
Penninger JM, Kroemer G (2003) Mitochondria, AIF and caspases—rivaling for cell death execution. Nat Cell Biol 5:97–99
Yip KW, Reed JC (2008) Bcl-2 family proteins and cancer. Oncogene 27:6398–6406
Kohn KW, Waring MJ, Glaubiger D, Friedman CA (1975) Intercalative binding of ellipticine to DNA. Cancer Res 35:71–76
Lo Y-S, Tseng W-H, Chuang C-Y, Hou M-H (2013) The structural basis of actinomycin D-binding induces nucleotide flipping out, a sharp bend and a left-handed twist in CGG triplet repeats. Nucleic Acids Res 41:4284–4294
Hengartner MO (2000) The biochemistry of apoptosis. Nature 407:770–776
Gavrilescu LC, Denkers EY (2003) Apoptosis and the balance of homeostatic and pathologic responses to protozoan infection. Infect Immun 71:6109–6115
Cheah YH, Nordin FJ, Sarip R, Tee TT, Azimahtol HLP, Sirat HM, Rashid BAA, Abdullah NR, Ismail Z (2009) Combined xanthorrhizol-curcumin exhibits synergistic growth inhibitory activity via apoptosis induction in human breast cancer cells MDA-MB-231. Cancer Cell Int 9:1
Fleischer A, Ghadiri A, Dessauge F, Duhamel M, Rebollo MP, Alvarez-Franco F, Rebollo A (2006) Modulating apoptosis as a target for effective therapy. Mol Immunol 43:1065–1079
Ralph RK, Marshall B, Darkin S (1982) Anti-cancer drugs which intercalate into DNA: how do they act? Trends Biochem Sci 8:212–214
Wu MH, Yung BY (1994) Cell cycle phase-dependent cytotoxicity of actinomycin D in HeLa cells. Eur J Pharmacol 270:203–212
Kroemer G, Galluzzi L, Brenner C (2007) Mitochondrial membrane permeabilization in cell death. Physiol Rev 87:99–163
Thornberry NA, Lazebnik Y (1998) Caspases: enemies within. Science 281:1312–1316
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.
Conflict of interest
The authors declare no conflicts of interest.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Supplementary material 1 (DOCX 343 kb)
About this article
Cite this article
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
- DNA intercalators