Lower antioxidative capacity of multidrug-resistant cancer cells confers collateral sensitivity to protoflavone derivatives
Multidrug resistance (MDR) may develop due to a series of adaptive responses under a new stress conditions, such as chemotherapy. Novel strategies are urgently needed to fight MDR in cancer, and chemotherapeutics that are selective for MDR cancer cells offer a promising approach. Certain protoflavones were previously found to have potential in this regard.
Cytotoxicity of six protoflavones was assessed in different P-glycoprotein overexpressing MDR cancer cell lines and in their non-MDR counterparts. The impacts of compound 5, 6-methylprotoflavone previously published and a new derivative, 6-bromoprotoflavone (compound 6), on the cell cycle distribution were evaluated, and 6 was also studied for its potential to regulate the intracellular antioxidative capacity.
Protoflavones showed a significant cytotoxicity against all cancer cell lines and selectivity toward MDR cancer cells adapted to conventional chemotherapeutics. Inverse sensitivity versus MDR selectivity pattern was observed. Treatment with H2O2 showed that MDR cancer cells are more vulnerable to oxidative stress. Compounds 5 and 6 significantly decreased the portion of MDR cells in the G1 phase. The levels of reactive oxygen and nitrogen species (ROS/RNS) between MDR and non-MDR cells significantly differed upon exposure to 6, accompanied by changes in the glutathione (GSH) levels and in the expression of manganese superoxide dismutase (MnSOD), glutathione-S-transferase π (GST π) and hypoxia-inducible factor-1α (HIF-1α).
Our results suggest that MDR cancer cells can be more vulnerable to the pro-oxidative activity of protoflavones due to an impaired antioxidative defense that might arise during the adaptation processes provoked by chemotherapy.
KeywordsMultidrug resistance P-glycoprotein Protoflavones Pro-oxidative activity Reactive oxygen species Collateral sensitivity
This research was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia (Grant No III41031). The authors acknowledge the support from the National Science Council (NSC), Taiwan, the Szeged Foundation for Cancer Research and the Fundação para a Ciência e a Tecnologia (FCT), Portugal (PEsT-OE/SAU/UI0074/2011 and PEsT-OE/SAU/UI0074/2014). This work was performed within the framework of COST Actions CM1106 (Chemical Approaches to Targeting Drug Resistance in Cancer Stem Cells; providing an STSM grant to A. Martins) and CM1407 (Challenging organic syntheses inspired by nature—from natural products chemistry to drug discovery). A bilateral mobility grant provided by the Hungarian Academy of Sciences and the NSC, Taiwan (MOST 104-2911-I-037-501), is also acknowledged.
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Conflict of interest
- 1.Hall MD, Marshall TS, Kwit AD, Miller Jenkins LM, Dulcey AE, Madigan JP, Pluchino KM, Goldsborough AS, Brimacombe KR, Griffiths GL, Gottesman MM (2014) Inhibition of glutathione peroxidase mediates the collateral sensitivity of multidrug-resistant cells to tiopronin. J Biol Chem 289(31):21473–21489. doi: 10.1074/jbc.M114.581702 PubMedCentralCrossRefPubMedGoogle Scholar
- 6.Szakacs G, Hall MD, Gottesman MM, Boumendjel A, Kachadourian R, Day BJ, Baubichon-Cortay H, Di Pietro A (2014) Targeting the Achilles heel of multidrug-resistant cancer by exploiting the fitness cost of resistance. Chem Rev 114(11):5753–5774. doi: 10.1021/cr4006236 PubMedCentralCrossRefPubMedGoogle Scholar
- 9.Chen WY, Hsieh YA, Tsai CI, Kang YF, Chang FR, Wu YC, Wu CC (2011) Protoapigenone, a natural derivative of apigenin, induces mitogen-activated protein kinase-dependent apoptosis in human breast cancer cells associated with induction of oxidative stress and inhibition of glutathione S-transferase pi. Invest New Drugs 29(6):1347–1359. doi: 10.1007/s10637-010-9497-0 CrossRefPubMedGoogle Scholar
- 10.Wang HC, Lee AY, Chou WC, Wu CC, Tseng CN, Liu KY, Lin WL, Chang FR, Chuang DW, Hunyadi A, Wu YC (2012) Inhibition of ATR-dependent signaling by protoapigenone and its derivative sensitizes cancer cells to interstrand cross-link-generating agents in vitro and in vivo. Mol Cancer Ther 11(7):1443–1453. doi: 10.1158/1535-7163.MCT-11-0921 CrossRefPubMedGoogle Scholar
- 15.Hunyadi A, Chuang DW, Danko B, Chiang MY, Lee CL, Wang HC, Wu CC, Chang FR, Wu YC (2011) Direct semi-synthesis of the anticancer lead-drug protoapigenone from apigenin, and synthesis of further new cytotoxic protoflavone derivatives. PLoS ONE 6(8):e23922. doi: 10.1371/journal.pone.0023922 PubMedCentralCrossRefPubMedGoogle Scholar
- 20.NicAmhlaoibh R, Heenan M, Cleary I, Touhey S, O’Loughlin C, Daly C, Nunez G, Scanlon KJ, Clynes M (1999) Altered expression of mRNAs for apoptosis-modulating proteins in a low level multidrug resistant variant of a human lung carcinoma cell line that also expresses mdr1 mRNA. Int J Cancer 82(3):368–376CrossRefPubMedGoogle Scholar
- 24.Wang M, Kirk JS, Venkataraman S, Domann FE, Zhang HJ, Schafer FQ, Flanagan SW, Weydert CJ, Spitz DR, Buettner GR, Oberley LW (2005) Manganese superoxide dismutase suppresses hypoxic induction of hypoxia-inducible factor-1alpha and vascular endothelial growth factor. Oncogene 24(55):8154–8166. doi: 10.1038/sj.onc.1208986 PubMedGoogle Scholar
- 25.Podolski-Renic A, Jadranin M, Stankovic T, Bankovic J, Stojkovic S, Chiourea M, Aljancic I, Vajs V, Tesevic V, Ruzdijic S, Gagos S, Tanic N, Pesic M (2013) Molecular and cytogenetic changes in multi-drug resistant cancer cells and their influence on new compounds testing. Cancer Chemother Pharmacol 72(3):683–697. doi: 10.1007/s00280-013-2247-1 CrossRefPubMedGoogle Scholar
- 26.Chen HM, Chang FR, Hsieh YC, Cheng YJ, Hsieh KC, Tsai LM, Lin AS, Wu YC, Yuan SS (2011) A novel synthetic protoapigenone analogue, WYC02-9, induces DNA damage and apoptosis in DU145 prostate cancer cells through generation of reactive oxygen species. Free Radic Biol Med 50(9):1151–1162. doi: 10.1016/j.freeradbiomed.2011.01.015 CrossRefPubMedGoogle Scholar
- 27.Chiu CC, Chang HW, Chuang DW, Chang FR, Chang YC, Cheng YS, Tsai MT, Chen WY, Lee SS, Wang CK, Chen JY, Wang HM, Chen CC, Liu YC, Wu YC (2009) Fern plant-derived protoapigenone leads to DNA damage, apoptosis, and G(2)/m arrest in lung cancer cell line H1299. DNA Cell Biol 28(10):501–506. doi: 10.1089/dna.2009.0852 CrossRefPubMedGoogle Scholar
- 29.Liu Z, Yuan Q, Zhang X, Xiong C, Xue P, Ruan J (2012) RY10-4, a novel anti-tumor compound, exhibited its anti-angiogenesis activity by down-regulation of the HIF-1alpha and inhibition phosphorylation of AKT and mTOR. Cancer Chemother Pharmacol 69(6):1633–1640. doi: 10.1007/s00280-012-1873-3 CrossRefPubMedGoogle Scholar
- 32.Lorendeau D, Dury L, Genoux-Bastide E, Lecerf-Schmidt F, Simoes-Pires C, Carrupt PA, Terreux R, Magnard S, Di Pietro A, Boumendjel A, Baubichon-Cortay H (2014) Collateral sensitivity of resistant MRP1-overexpressing cells to flavonoids and derivatives through GSH efflux. Biochem Pharmacol 90(3):235–245. doi: 10.1016/j.bcp.2014.05.017 CrossRefPubMedGoogle Scholar
- 34.Chang HL, Wu YC, Su JH, Yeh YT, Yuan SS (2008) Protoapigenone, a novel flavonoid, induces apoptosis in human prostate cancer cells through activation of p38 mitogen-activated protein kinase and c-Jun NH2-terminal kinase 1/2. J Pharmacol Exp Ther 325(3):841–849. doi: 10.1124/jpet.107.135442 CrossRefPubMedGoogle Scholar
- 35.Chen YJ, Kay N, Yang JM, Lin CT, Chang HL, Wu YC, Fu CF, Chang Y, Lo S, Hou MF, Lee YC, Hsieh YC, Yuan SS (2013) Total synthetic protoapigenone WYC02 inhibits cervical cancer cell proliferation and tumour growth through PIK3 signalling pathway. Basic Clin Pharmacol Toxicol 113(1):8–18. doi: 10.1111/bcpt.12057 CrossRefPubMedGoogle Scholar
- 37.Chen J (2014) Reactive oxygen species and drug resistance in cancer chemotherapy. Austin J Clin Pathol 1(4):1017Google Scholar