The aim of this work was to study the antitumor effects and the mechanisms of toxic action of a series of 6-methoxyquinoline (6MQ) complexes in vitro. The Cu(II) and Zn(II) complexes (Cu6MQ and Zn6MQ) are formulated as M(6MQ)2Cl2; the Co(II) and Ag(I) compounds (Co6MQ and Ag6MQ) are ionic with formulae [Ag(6MQ)2]+NO3− and H(6MQ)+[Co(6MQ)Cl3]− (where H(6MQ)+ is the protonated ligand). We found that the copper complex, outperformed the Co(II), Zn(II) and Ag(I) complexes with a lower IC50 (57.9 µM) in A549 cells exposed for 24 h. Cu6MQ decreased cell proliferation and induced oxidative stress detected with H2DCFDA at 40 µM, which reduces GSH/GSSG ratio. This redox imbalance induced oxidative DNA damage revealed by the Micronucleus test and the Comet assay, which turned into a cell cycle arrest at G2/M phase and induced apoptosis. In multicellular spheroids, the IC50 values tripled the monolayer model (187.3 µM for 24 h). At this concentration, the proportion of live/dead cells diminished, and the spheroids could not proliferate or invade. Although Zn6MQ also decreased GSH/GSSG ratio from 200 µM and the cytotoxicity is related to oxidative stress, the induction of the hydrogen peroxide levels only doubled the control value. Zn6MQ induced S phase arrest, which relates with the increased micronucleus frequency and with the induction of necrosis. Finally, our results reveal a synergistic activity with a 1:1 ratio of both complexes in the monolayer and multicellular spheroids.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Liberti MV, Locasale JW (2016) The Warburg effect: how does it benefit cancer cells? Trends Biochem Sci 41:211–218. https://doi.org/10.1016/j.tibs.2015.12.001
Li P, Zhang D, Shen L et al (2016) Redox homeostasis protects mitochondria through accelerating ROS conversion to enhance hypoxia resistance in cancer cells. Sci Rep 6:1–13. https://doi.org/10.1038/srep22831
Laurent A, Nicco C, Chéreau C et al (2005) Controlling tumor growth by modulating endogenous production of reactive oxygen species. Cancer Res 65:948–956
Lin Y, Zhang H, Liang J et al (2014) Identification and characterization of alphavirus M1 as a selective oncolytic virus targeting ZAP-defective human cancers. Proc Natl Acad Sci USA. https://doi.org/10.1073/pnas.1408759111
Surova O, Zhivotovsky B (2013) Various modes of cell death induced by DNA damage. Oncogene 32:3789–3797. https://doi.org/10.1038/onc.2012.556
Ceccacci E, Minucci S (2016) Inhibition of histone deacetylases in cancer therapy: lessons from leukaemia. Br J Cancer 114:605–611. https://doi.org/10.1038/bjc.2016.36
Butler LM, Zhou X, Xu W-S et al (2002) The histone deacetylase inhibitor SAHA arrests cancer cell growth, up-regulates thioredoxin-binding protein-2, and down-regulates thioredoxin. Proc Natl Acad Sci 99:11700–11705. https://doi.org/10.1073/pnas.182372299
Dawson MA, Kouzarides T (2012) Cancer epigenetics: from mechanism to therapy. Cell 150:12–27. https://doi.org/10.1016/j.cell.2012.06.013
Eot-Houllier G, Fulcrand G, Magnaghi-Jaulin L, Jaulin C (2009) Histone deacetylase inhibitors and genomic instability. Cancer Lett 274:169–176. https://doi.org/10.1016/j.canlet.2008.06.005
Martirosyan AR, Rahim-Bata R, Freeman AB et al (2004) Differentiation-inducing quinolines as experimental breast cancer agents in the MCF-7 human breast cancer cell model. Biochem Pharmacol 68:1729–1738. https://doi.org/10.1016/J.BCP.2004.05.003
Lee H-Y, Nepali K, Huang F-I et al (2018) (N-Hydroxycarbonylbenylamino)quinolines as selective histone deacetylase 6 inhibitors suppress growth of multiple myeloma in vitro and in vivo. J Med Chem 61:905–917. https://doi.org/10.1021/acs.jmedchem.7b01404
Arafa RK, Hegazy GH, Piazza GA, Abadi AH (2013) Synthesis and in vitro antiproliferative effect of novel quinoline-based potential anticancer agents. Eur J Med Chem 63:826–832. https://doi.org/10.1016/j.ejmech.2013.03.008
Nunoshiba T, Demple B (1993) Potent intracellular oxidative stress exerted by the carcinogen 4-nitroquinoline-N-oxide. Cancer Res 53:3250–3252
Kwon S, Lee Y, Jung Y et al (2018) Mitochondria-targeting indolizino[3,2-c]quinolines as novel class of photosensitizers for photodynamic anticancer activity. Eur J Med Chem 148:116–127. https://doi.org/10.1016/J.EJMECH.2018.02.016
Allan JR, Dahyrnple J (1991) Thermal, spectral and magnetic studies of cobalt(II), copper(II) and zinc(II) complexes of 5,6-benzoquinoline and 6-methoxyquinoline. Thermochim Acta Elsevier Sci Publ BV 191:223–230
Villa-Pérez C, Oyarzabal I, Echeverría GA et al (2016) Single-ion magnets based on mononuclear cobalt(II) complexes with sulfadiazine. Eur J Inorg Chem 2016:4835–4841. https://doi.org/10.1002/ejic.201600777
Villa-Pérez C, Ortega IC, Vélez-Macías A et al (2018) Crystal structure, physicochemical properties, Hirshfeld surface analysis and antibacterial activity assays of transition metal complexes of 6-methoxyquinoline. New J Chem. https://doi.org/10.1039/c8nj00661j
Friedrich J, Ebner R, Kunz-Schughart LA (2007) Experimental anti-tumor therapy in 3-D: spheroids—old hat or new challenge? Int J Radiat Biol 83:849–871. https://doi.org/10.1080/09553000701727531
Friedrich J, Eder W, Castaneda J et al (2007) A reliable tool to determine cell viability in complex 3-D culture: the acid phosphatase assay. J Biomol Screen 12:925–937. https://doi.org/10.1177/1087057107306839
Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63
Franken NAP, Rodermond HM, Stap J et al (2006) Clonogenic assay of cells in vitro. Nat Protoc 1:2315–2319. https://doi.org/10.1038/nprot.2006.339
Hissin PJ, Hilf R (1976) A fluorometric method for determination of oxidized and reduced glutathione in tissues. Anal Biochem 74:214–226
Fenech M (2000) The in vitro micronucleus technique. Mutat Res Mol Mech Mutagen 455:81–95. https://doi.org/10.1016/S0027-5107(00)00065-8
Anoopkumar-Dukie S, Carey JB, Conere T et al (2005) Resazurin assay of radiation response in cultured cells. Br J Radiol 78:945–947. https://doi.org/10.1259/bjr/54004230
Munshi A, Hobbs M, Meyn RE (2005) Clonogenic cell survival assay. Methods Mol Med 110:21–28. https://doi.org/10.1385/1-59259-869-2:021
Jemal A, Bray F, Center MM et al (2011) Global cancer statistics. CA Cancer J Clin 61:69–90. https://doi.org/10.3322/caac.20107
Torre LA, Bray F, Siegel RL et al (2015) Global cancer statistics, 2012. CA Cancer J Clin 65:87–108. https://doi.org/10.3322/caac.21262
Hirsch FR, Scagliotti GV, Mulshine JL et al (2017) Lung cancer: current therapies and new targeted treatments. Lancet 389:299–311. https://doi.org/10.1016/S0140-6736(16)30958-8
Novello S, Barlesi F, Califano R et al (2016) Metastatic non-small-cell lung cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol 27:V1–V27. https://doi.org/10.1093/annonc/mdw326
Damaskos C, Tomos I, Garmpis N et al (2018) Histone deacetylase inhibitors as a novel targeted therapy against non-small cell lung cancer: where are we now and what should we expect? Anticancer Res 38:37–43. https://doi.org/10.21873/anticanres.12189
Yu W, Lu W, Chen G et al (2017) Inhibition of histone deacetylases sensitizes EGF receptor-TK inhibitor-resistant non-small-cell lung cancer cells to erlotinib in vitro and in vivo. Br J Pharmacol 174:3608–3622. https://doi.org/10.1111/bph.13961
Wang L, Li H, Ren Y et al (2016) Targeting HDAC with a novel inhibitor effectively reverses paclitaxel resistance in non-small cell lung cancer via multiple mechanisms. Cell Death Dis 7:e2063. https://doi.org/10.1038/cddis.2015.328
Rogolino D, Cavazzoni A, Gatti A et al (2017) Anti-proliferative effects of copper(II) complexes with hydroxyquinoline-thiosemicarbazone ligands. Eur J Med Chem 128:140–153. https://doi.org/10.1016/J.EJMECH.2017.01.031
Angel NR, Khatib RM, Jenkins J et al (2017) Copper (II) complexes possessing alkyl-substituted polypyridyl ligands: structural characterization and in vitro antitumor activity. J Inorg Biochem 166:12–25. https://doi.org/10.1016/J.JINORGBIO.2016.09.012
Stanojkovic TP, Kovala-Demertzi D, Primikyri A et al (2010) Zinc(II) complexes of 2-acetyl pyridine 1-(4-fluorophenyl)-piperazinyl thiosemicarbazone: synthesis, spectroscopic study and crystal structures—potential anticancer drugs. J Inorg Biochem 104:467–476. https://doi.org/10.1016/J.JINORGBIO.2009.12.021
Casas JS, Castellano EE, Couce MD et al (2006) Zinc(II), cadmium(II) and mercury(II) complexes of the vitamin B1 antagonist oxythiamine. J Inorg Biochem 100:124–132. https://doi.org/10.1016/J.JINORGBIO.2005.10.009
Cadavid-Vargas JFJ, León IE, Etcheverry SSB et al (2017) Copper(II) complexes with saccharinate and glutamine as antitumor agents: cytoand genotoxicity in human osteosarcoma cells. Anticancer Agents Med Chem 17:424–433. https://doi.org/10.2174/1871520616666160513130204
Karlsson H, Fryknäs M, Strese S et al (2017) Mechanistic characterization of a copper containing thiosemicarbazone with potent antitumor activity. Oncotarget 8:30217–30234. https://doi.org/10.18632/oncotarget.16324
Subastri A, Suyavaran A, Preedia Babu E et al (2018) Troxerutin with copper generates oxidative stress in cancer cells: its possible chemotherapeutic mechanism against hepatocellular carcinoma. J Cell Physiol 233:1775–1790. https://doi.org/10.1002/jcp.26061
Martínez VR, Aguirre MV, Todaro JS et al (2018) Azilsartan and its Zn(II) complex. Synthesis, anticancer mechanisms of action and binding to bovine serum albumin. Toxicol Vitr 48:205–220. https://doi.org/10.1016/J.TIV.2018.01.009
Tan YS, Ooi KK, Ang KP et al (2015) Molecular mechanisms of apoptosis and cell selectivity of zinc dithiocarbamates functionalized with hydroxyethyl substituents. J Inorg Biochem 150:48–62. https://doi.org/10.1016/J.JINORGBIO.2015.06.009
Mohammadizadeh F, Falahati-pour SK, Rezaei A et al (2018) The cytotoxicity effects of a novel Cu complex on MCF-7 human breast cancerous cells. Biometals 31:233–242. https://doi.org/10.1007/s10534-018-0079-5
Gouda AM, El-Ghamry HA, Bawazeer TM et al (2018) Antitumor activity of pyrrolizines and their Cu(II) complexes: design, synthesis and cytotoxic screening with potential apoptosis-inducing activity. Eur J Med Chem 145:350–359. https://doi.org/10.1016/J.EJMECH.2018.01.009
Portugal J, Mansilla S, Bataller M (2010) Mechanisms of drug-induced mitotic catastrophe in cancer cells. Curr Pharm Des 16:69–78. https://doi.org/10.2174/138161210789941801
Khabour OF, Saleh N, Alzoubi KH et al (2013) Genotoxicity of structurally related copper and zinc containing Schiff base complexes. Drug Chem Toxicol 36:435–442. https://doi.org/10.3109/01480545.2013.776577
Leon I, Cadavid-Vargas J, Di Virgilio A, Etcheverry S (2017) Vanadium, ruthenium and copper compounds: a new class of nonplatinum metallodrugs with anticancer activity. Curr Med Chem 24:112–148. https://doi.org/10.2174/0929867323666160824162546
Santini C, Pellei M, Gandin V et al (2014) Advances in copper complexes as anticancer agents. Chem Rev 114:815–862. https://doi.org/10.1021/cr400135x
Serment-Guerrero J, Bravo-Gomez ME, Lara-Rivera E, Ruiz-Azuara L (2017) Genotoxic assessment of the copper chelated compounds Casiopeinas: clues about their mechanisms of action. J Inorg Biochem 166:68–75. https://doi.org/10.1016/J.JINORGBIO.2016.11.007
Rhaese H-J, Freese E (1968) Chemical analysis of DNA alterations: I. Base liberation and backbone breakage of DNA and oligodeoxyadenylic acid induced by hydrogen peroxide and hydroxylamine. Biochim Biophys Acta Nucleic Acids Protein Synth 155:476–490. https://doi.org/10.1016/0005-2787(68)90193-7
Adhikari A, Kumari N, Adhikari M et al (2017) Zinc complex of tryptophan appended 1,4,7,10-tetraazacyclododecane as potential anticancer agent: synthesis and evaluation. Bioorg Med Chem 25:3483–3490. https://doi.org/10.1016/J.BMC.2017.04.035
Santra M, Das SK, Talukder G, Sharma A (2002) Induction of micronuclei by zinc in human leukocytes. Biol Trace Elem Res 88:139–144. https://doi.org/10.1385/BTER:88:2:139
Scicchitano DA, Pegg AE (1987) Inhibition of O6-alkylguanine-DNA-alkyltransferase by metals. Mutat Res Lett 192:207–210. https://doi.org/10.1016/0165-7992(87)90057-1
Yang SW, Becker FF, Chan JYH (1996) Inhibition of human DNA ligase I activity by zinc and cadmium and the fidelity of ligation. Environ Mol Mutagen 28:19–25. https://doi.org/10.1002/(SICI)1098-2280(1996)28:1%3c19:AID-EM5%3e3.0.CO;2-9
Xu B, Sun Z, Liu Z et al (2011) Replication stress induces micronuclei comprising of aggregated DNA double-strand breaks. PLoS One. https://doi.org/10.1371/journal.pone.0018618
Galateanu B, Hudita A, Negrei C et al (2016) Impact of multicellular tumor spheroids as an in vivo-like tumor model on anticancer drug response. Int J Oncol 48:2295–2302. https://doi.org/10.3892/ijo.2016.3467
Shi X, Chen Z, Wang Y et al (2018) Hypotoxic copper complexes with potent anti-metastatic and anti-angiogenic activities against cancer cells. Dalt Trans 47:5049–5054. https://doi.org/10.1039/C8DT00794B
Tallarida RJ (2001) Drug synergism: its detection and applications. J Pharmacol Exp Ther 298:865–872
Marcato-Romain CE, Pinelli E, Pourrut B et al (2009) Assessment of the genotoxicity of Cu and Zn in raw and anaerobically digested slurry with the Vicia faba micronucleus test. Mutat Res Genet Toxicol Environ Mutagen 672:113–118. https://doi.org/10.1016/j.mrgentox.2008.10.018
This work was supported by UNLP (11X/690, PPID 2018/X032), CONICET (PIP 0034) and ANPCyT (PICT 2014-2223 and PICT 2016-0508) from Argentina.
Conflict of interest
The authors confirm that they have no conflict of interest with the content of this article.
This article does not contain studies with human participants or animals performed by any of the authors.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
About this article
Cite this article
Cadavid-Vargas, J.F., Villa-Pérez, C., Ruiz, M.C. et al. 6-Methoxyquinoline complexes as lung carcinoma agents: induction of oxidative damage on A549 monolayer and multicellular spheroid model. J Biol Inorg Chem 24, 271–285 (2019). https://doi.org/10.1007/s00775-019-01644-7
- 6-Methoxyquinoline complexes
- Lung carcinoma
- A549 cells
- Multicellular spheroid model
- Oxidative damage