{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,31]],"date-time":"2025-12-31T12:18:58Z","timestamp":1767183538296},"reference-count":80,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2018,12,10]],"date-time":"2018-12-10T00:00:00Z","timestamp":1544400000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2018,12,10]],"date-time":"2018-12-10T00:00:00Z","timestamp":1544400000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>In the present study, we examined a hypothesis that dichloroacetate, a metabolic inhibitor, might efficiently potentiate the cytotoxic effect of salinomycin, an antibiotic ionophore, on two human colorectal cancer derived cell lines DLD-1 and HCT116. First, we performed a series of dose response experiments in the 2D cell culture by applying mono- and combination therapy and by using the Chou-Talalay method found that salinomycin in combination with dichloroacetate acted synergistically in both cell lines. Secondly, in order to recapitulate the <jats:italic>in vivo<\/jats:italic> tumor architecture, we tested various doses of these compounds, alone and in combination, in the 3D multicellular spheroid culture. The effect of combination of dichloracetate and salinomycin on multicellular spheroid size was stronger than the sum of both monotherapies, particularly in HCT116 cells. Further, we demonstrate that the synergistic effect of compounds may be related to the inhibitory effect of dichloroacetate on multidrug resistance proteins, and in contrast, it is not related to dichloroacetate-induced reduction of intracellular pH. Our findings indicate that the combination therapy of salinomycin and dichloroacetate could be an effective option for colorectal cancer treatment and provide the first mechanistic explanation of the synergistic action of these compounds.<\/jats:p>","DOI":"10.1038\/s41598-018-35815-4","type":"journal-article","created":{"date-parts":[[2018,12,4]],"date-time":"2018-12-04T13:26:46Z","timestamp":1543930006000},"update-policy":"http:\/\/dx.doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Dichloroacetate and Salinomycin Exert a Synergistic Cytotoxic Effect in Colorectal Cancer Cell Lines"],"prefix":"10.1038","volume":"8","author":[{"given":"Aist\u0117","family":"Skeberdyt\u0117","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ieva","family":"Sarapinien\u0117","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jan","family":"Aleksander-Krasko","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vaidotas","family":"Stankevi\u010dius","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"K\u0119stutis","family":"Su\u017eied\u0117lis","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sonata","family":"Jarmalait\u0117","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2018,12,10]]},"reference":[{"key":"35815_CR1","first-page":"105","volume":"7","author":"AR Marley","year":"2016","unstructured":"Marley, A. R. & Nan, H. Epidemiology of colorectal cancer. Int J Mol Epidemiol Genet 7, 105\u2013114 (2016).","journal-title":"Int J Mol Epidemiol Genet"},{"key":"35815_CR2","first-page":"146","volume":"1","author":"JL Marshall","year":"2007","unstructured":"Marshall, J. L. et al. Adjuvant Therapy for Stage II and III Colon Cancer: Consensus Report of the International Society of Gastrointestinal Oncology. Gastrointest Cancer Res 1, 146\u2013154 (2007).","journal-title":"Gastrointest Cancer Res"},{"key":"35815_CR3","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1186\/1878-5085-4-3","volume":"4","author":"S Hagan","year":"2016","unstructured":"Hagan, S., Orr, M. C. & Doyle, B. Targeted therapies in colorectal cancer-an integrative view by PPPM. Epma J 4, 3 (2016).","journal-title":"Epma J"},{"key":"35815_CR4","doi-asserted-by":"publisher","first-page":"229","DOI":"10.1200\/JCO.2004.05.113","volume":"22","author":"C Tournigand","year":"2004","unstructured":"Tournigand, C. et al. FOLFIRI followed by FOLFOX6 or the reverse sequence in advanced colorectal cancer: a randomized GERCOR study. J Clin Oncol 22, 229\u2013237 (2004).","journal-title":"J Clin Oncol"},{"key":"35815_CR5","doi-asserted-by":"publisher","first-page":"3399","DOI":"10.1038\/onc.2015.398","volume":"35","author":"M Oliver Metzig","year":"2016","unstructured":"Oliver Metzig, M. et al. Inhibition of caspases primes colon cancer cells for 5-fluorouracil-induced TNF-alpha-dependent necroptosis driven by RIP1 kinase and NF-kappaB. Oncogene 35, 3399\u20133409 (2016).","journal-title":"Oncogene"},{"key":"35815_CR6","doi-asserted-by":"publisher","first-page":"330","DOI":"10.1038\/nrc1074","volume":"3","author":"DB Longley","year":"2003","unstructured":"Longley, D. B., Harkin, D. P. & Johnston, P. G. 5-fluorouracil: mechanisms of action and clinical strategies. Nat Rev Cancer 3, 330\u2013338 (2003).","journal-title":"Nat Rev Cancer"},{"key":"35815_CR7","doi-asserted-by":"crossref","first-page":"38022","DOI":"10.18632\/oncotarget.16723","volume":"8","author":"R Bayat Mokhtari","year":"2017","unstructured":"Bayat Mokhtari, R. et al. Combination therapy in combating cancer. Oncotarget 8, 38022\u201338043 (2017).","journal-title":"Oncotarget"},{"key":"35815_CR8","doi-asserted-by":"publisher","first-page":"645","DOI":"10.1016\/j.cell.2009.06.034","volume":"138","author":"PB Gupta","year":"2009","unstructured":"Gupta, P. B. et al. Identification of selective inhibitors of cancer stem cells by high-throughput screening. Cell 138, 645\u2013659 (2009).","journal-title":"Cell"},{"key":"35815_CR9","doi-asserted-by":"publisher","first-page":"1898","DOI":"10.3892\/mmr.2015.3633","volume":"12","author":"KY Kim","year":"2015","unstructured":"Kim, K. Y. et al. Salinomycin enhances doxorubicin-induced cytotoxicity in multidrug resistant MCF-7\/MDR human breast cancer cells via decreased efflux of doxorubicin. Mol Med Rep 12, 1898\u20131904 (2015).","journal-title":"Mol Med Rep"},{"key":"35815_CR10","doi-asserted-by":"publisher","first-page":"696","DOI":"10.1016\/j.bbrc.2015.09.108","volume":"466","author":"H An","year":"2015","unstructured":"An, H. et al. Salinomycin possesses anti-tumor activity and inhibits breast cancer stem-like cells via an apoptosis-independent pathway. Biochem Biophys Res Commun 466, 696\u2013703 (2015).","journal-title":"Biochem Biophys Res Commun"},{"key":"35815_CR11","doi-asserted-by":"publisher","DOI":"10.1038\/cddis.2013.223","volume":"4","author":"KH Koo","year":"2013","unstructured":"Koo, K. H. et al. Salinomycin induces cell death via inactivation of Stat3 and downregulation of Skp2. Cell Death Dis 4, e693 (2013).","journal-title":"Cell Death Dis"},{"key":"35815_CR12","first-page":"1020","volume":"2","author":"S Zhou","year":"2014","unstructured":"Zhou, S. et al. Salinomycin Suppresses PDGFRbeta, MYC, and Notch Signaling in Human Medulloblastoma. Austin J Pharmacol Ther 2, 1020 (2014).","journal-title":"Austin J Pharmacol Ther"},{"key":"35815_CR13","first-page":"4101653","volume":"2017","author":"Y Zhang","year":"2017","unstructured":"Zhang, Y. et al. Salinomycin Exerts Anticancer Effects on PC-3 Cells and PC-3-Derived Cancer Stem Cells In Vitro and In Vivo. Biomed Res Int 2017, 4101653 (2017).","journal-title":"Biomed Res Int"},{"key":"35815_CR14","doi-asserted-by":"publisher","first-page":"e14","DOI":"10.3802\/jgo.2017.28.e14","volume":"28","author":"HG Lee","year":"2017","unstructured":"Lee, H. G. et al. Salinomycin reduces stemness and induces apoptosis on human ovarian cancer stem cell. J Gynecol Oncol 28, e14 (2017).","journal-title":"J Gynecol Oncol"},{"issue":"12","key":"35815_CR15","doi-asserted-by":"publisher","first-page":"2808","DOI":"10.1002\/ijc.27595","volume":"131","author":"Prajakta S. Oak","year":"2012","unstructured":"Oak, P. S. et al. Combinatorial treatment of mammospheres with trastuzumab and salinomycin efficiently targets HER2-positive cancer cells and cancer stem cells. Int J Cancer 131, 2808\u20132819.","journal-title":"International Journal of Cancer"},{"key":"35815_CR16","doi-asserted-by":"publisher","first-page":"1098","DOI":"10.1016\/j.bbrc.2010.03.138","volume":"394","author":"D Fuchs","year":"2010","unstructured":"Fuchs, D., Daniel, V., Sadeghi, M., Opelz, G. & Naujokat, C. Salinomycin overcomes ABC transporter-mediated multidrug and apoptosis resistance in human leukemia stem cell-like KG-1a cells. Biochem Biophys Res Commun 394, 1098\u20131104 (2010).","journal-title":"Biochem Biophys Res Commun"},{"key":"35815_CR17","doi-asserted-by":"publisher","first-page":"13253","DOI":"10.1073\/pnas.1110431108","volume":"108","author":"D Lu","year":"2011","unstructured":"Lu, D. et al. Salinomycin inhibits Wnt signaling and selectively induces apoptosis in chronic lymphocytic leukemia cells. Proc Natl Acad Sci USA 108, 13253\u201313257 (2011).","journal-title":"Proc Natl Acad Sci USA"},{"key":"35815_CR18","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1016\/j.bcmd.2010.03.008","volume":"45","author":"R Riccioni","year":"2010","unstructured":"Riccioni, R. et al. The cancer stem cell selective inhibitor salinomycin is a p-glycoprotein inhibitor. Blood Cells Mol Dis 45, 86\u201392 (2010).","journal-title":"Blood Cells Mol Dis"},{"key":"35815_CR19","doi-asserted-by":"publisher","first-page":"655","DOI":"10.1128\/AAC.9.4.655","volume":"9","author":"M Mitani","year":"1976","unstructured":"Mitani, M., Yamanishi, T., Miyazaki, Y. & Otake, N. Salinomycin effects on mitochondrial ion translocation and respiration. Antimicrob Agents Chemother 9, 655\u2013660 (1976).","journal-title":"Antimicrob Agents Chemother"},{"key":"35815_CR20","doi-asserted-by":"publisher","first-page":"1658","DOI":"10.1158\/0008-5472.CAN-12-4188","volume":"73","author":"SF Pedersen","year":"2013","unstructured":"Pedersen, S. F. & Stock, C. Ion channels and transporters in cancer: pathophysiology, regulation, and clinical potential. Cancer Res 73, 1658\u20131661 (2013).","journal-title":"Cancer Res"},{"key":"35815_CR21","doi-asserted-by":"publisher","first-page":"80","DOI":"10.1016\/j.bbrc.2011.08.054","volume":"413","author":"KY Kim","year":"2011","unstructured":"Kim, K. Y. et al. Salinomycin-induced apoptosis of human prostate cancer cells due to accumulated reactive oxygen species and mitochondrial membrane depolarization. Biochem Biophys Res Commun 413, 80\u201386 (2011).","journal-title":"Biochem Biophys Res Commun"},{"key":"35815_CR22","doi-asserted-by":"publisher","first-page":"99","DOI":"10.1038\/bjc.2011.530","volume":"106","author":"K Ketola","year":"2012","unstructured":"Ketola, K. et al. Salinomycin inhibits prostate cancer growth and migration via induction of oxidative stress. Br J Cancer 106, 99\u2013106 (2012).","journal-title":"Br J Cancer"},{"key":"35815_CR23","doi-asserted-by":"crossref","unstructured":"Naujokat, C. & Steinhart, R. Salinomycin as a drug for targeting human cancer stem cells. J Biomed Biotechnol, 950658 (2012).","DOI":"10.1155\/2012\/950658"},{"key":"35815_CR24","doi-asserted-by":"publisher","first-page":"394","DOI":"10.1016\/j.ygyno.2008.01.038","volume":"109","author":"JY Wong","year":"2008","unstructured":"Wong, J. Y., Huggins, G. S., Debidda, M., Munshi, N. C. & De Vivo, I. Dichloroacetate induces apoptosis in endometrial cancer cells. Gynecol Oncol 109, 394\u2013402 (2008).","journal-title":"Gynecol Oncol"},{"key":"35815_CR25","doi-asserted-by":"publisher","first-page":"1223","DOI":"10.1002\/pros.20788","volume":"68","author":"W Cao","year":"2008","unstructured":"Cao, W. et al. Dichloroacetate (DCA) sensitizes both wild-type and over expressing Bcl-2 prostate cancer cells in vitro to radiation. Prostate 68, 1223\u20131231 (2008).","journal-title":"Prostate"},{"key":"35815_CR26","doi-asserted-by":"publisher","first-page":"1001","DOI":"10.1002\/ijc.25728","volume":"128","author":"I Papandreou","year":"2011","unstructured":"Papandreou, I., Goliasova, T. & Denko, N. C. Anticancer drugs that target metabolism: Is dichloroacetate the new paradigm? Int J Cancer 128, 1001\u20131008 (2011).","journal-title":"Int J Cancer"},{"key":"35815_CR27","doi-asserted-by":"publisher","first-page":"989","DOI":"10.1038\/sj.bjc.6604554","volume":"99","author":"ED Michelakis","year":"2008","unstructured":"Michelakis, E. D., Webster, L. & Mackey, J. R. Dichloroacetate (DCA) as a potential metabolic-targeting therapy for cancer. Br J Cancer 99, 989\u2013994 (2008).","journal-title":"Br J Cancer"},{"key":"35815_CR28","doi-asserted-by":"publisher","first-page":"2510","DOI":"10.1002\/ijc.25499","volume":"127","author":"LH Stockwin","year":"2010","unstructured":"Stockwin, L. H. et al. Sodium dichloroacetate selectively targets cells with defects in the mitochondrial ETC. Int J Cancer 127, 2510\u20132519 (2010).","journal-title":"Int J Cancer"},{"key":"35815_CR29","doi-asserted-by":"publisher","DOI":"10.1186\/1476-4598-10-142","volume":"10","author":"RC Sun","year":"2011","unstructured":"Sun, R. C., Board, P. G. & Blackburn, A. C. Targeting metabolism with arsenic trioxide and dichloroacetate in breast cancer cells. Mol Cancer 10, 142 (2011).","journal-title":"Mol Cancer"},{"key":"35815_CR30","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/j.ccr.2006.10.020","volume":"11","author":"S Bonnet","year":"2007","unstructured":"Bonnet, S. et al. A mitochondria-K+ channel axis is suppressed in cancer and its normalization promotes apoptosis and inhibits cancer growth. Cancer Cell 11, 37\u201351 (2007).","journal-title":"Cancer Cell"},{"key":"35815_CR31","doi-asserted-by":"publisher","first-page":"1124","DOI":"10.1016\/0026-0495(89)90051-6","volume":"38","author":"PW Stacpoole","year":"1989","unstructured":"Stacpoole, P. W. The pharmacology of dichloroacetate. Metabolism 38, 1124\u20131144 (1989).","journal-title":"Metabolism"},{"key":"35815_CR32","doi-asserted-by":"publisher","first-page":"1519","DOI":"10.1542\/peds.2005-1226","volume":"117","author":"PW Stacpoole","year":"2006","unstructured":"Stacpoole, P. W. et al. Controlled clinical trial of dichloroacetate for treatment of congenital lactic acidosis in children. Pediatrics 117, 1519\u20131531 (2006).","journal-title":"Pediatrics"},{"key":"35815_CR33","doi-asserted-by":"publisher","first-page":"253","DOI":"10.1007\/s10549-009-0435-9","volume":"120","author":"RC Sun","year":"2010","unstructured":"Sun, R. C. et al. Reversal of the glycolytic phenotype by dichloroacetate inhibits metastatic breast cancer cell growth in vitro and in vivo. Breast Cancer Res Treat 120, 253\u2013260 (2010).","journal-title":"Breast Cancer Res Treat"},{"key":"35815_CR34","doi-asserted-by":"publisher","first-page":"452","DOI":"10.1007\/s10637-013-0047-4","volume":"32","author":"EM Dunbar","year":"2014","unstructured":"Dunbar, E. M. et al. Phase 1 trial of dichloroacetate (DCA) in adults with recurrent malignant brain tumors. Invest New Drugs 32, 452\u2013464 (2014).","journal-title":"Invest New Drugs"},{"key":"35815_CR35","doi-asserted-by":"publisher","first-page":"603","DOI":"10.1007\/s10637-015-0221-y","volume":"33","author":"QS Chu","year":"2015","unstructured":"Chu, Q. S. et al. A phase I open-labeled, single-arm, dose-escalation, study of dichloroacetate (DCA) in patients with advanced solid tumors. Invest New Drugs 33, 603\u2013610 (2015).","journal-title":"Invest New Drugs"},{"key":"35815_CR36","doi-asserted-by":"publisher","first-page":"31ra34","DOI":"10.1126\/scitranslmed.3000677","volume":"2","author":"ED Michelakis","year":"2010","unstructured":"Michelakis, E. D. et al. Metabolic modulation of glioblastoma with dichloroacetate. Sci Transl Med 2, 31ra34 (2010).","journal-title":"Sci Transl Med"},{"key":"35815_CR37","doi-asserted-by":"publisher","first-page":"255","DOI":"10.1007\/s11060-017-2664-9","volume":"136","author":"M Albatany","year":"2018","unstructured":"Albatany, M., Li, A., Meakin, S. & Bartha, R. Dichloroacetate induced intracellular acidification in glioblastoma: in vivo detection using AACID-CEST MRI at 9.4 Tesla. J Neurooncol 136, 255\u2013262 (2018).","journal-title":"J Neurooncol"},{"key":"35815_CR38","doi-asserted-by":"publisher","first-page":"541","DOI":"10.1016\/S0305-7372(03)00106-3","volume":"29","author":"H Izumi","year":"2003","unstructured":"Izumi, H. et al. Cellular pH regulators: potentially promising molecular targets for cancer chemotherapy. Cancer Treat Rev 29, 541\u2013549 (2003).","journal-title":"Cancer Treat Rev"},{"key":"35815_CR39","doi-asserted-by":"publisher","first-page":"767","DOI":"10.1038\/nrd3554","volume":"10","author":"D Neri","year":"2011","unstructured":"Neri, D. & Supuran, C. T. Interfering with pH regulation in tumours as a therapeutic strategy. Nat Rev Drug Discov 10, 767\u2013777 (2011).","journal-title":"Nat Rev Drug Discov"},{"key":"35815_CR40","doi-asserted-by":"publisher","first-page":"621","DOI":"10.1124\/pr.58.3.10","volume":"58","author":"TC Chou","year":"2006","unstructured":"Chou, T. C. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev 58, 621\u2013681 (2006).","journal-title":"Pharmacol Rev"},{"key":"35815_CR41","doi-asserted-by":"publisher","first-page":"2071","DOI":"10.1021\/mp500002y","volume":"11","author":"TM Achilli","year":"2014","unstructured":"Achilli, T. M., McCalla, S., Meyer, J., Tripathi, A. & Morgan, J. R. Multilayer spheroids to quantify drug uptake and diffusion in 3D. Mol Pharm 11, 2071\u20132081 (2014).","journal-title":"Mol Pharm"},{"issue":"2","key":"35815_CR42","doi-asserted-by":"publisher","first-page":"215","DOI":"10.1016\/j.bbamem.2005.01.014","volume":"1711","author":"Juan C. S\u00e1ez","year":"2005","unstructured":"Saez, J. C., Retamal, M. A., Basilio, D., Bukauskas, F. F. & Bennett, M. V. Connexin-based gap junction hemichannels: gating mechanisms. Biochim Biophys Acta. 1711, 215\u2013224. Epub 2005 Mar 2002. (2005).","journal-title":"Biochimica et Biophysica Acta (BBA) - Biomembranes"},{"issue":"5","key":"35815_CR43","doi-asserted-by":"publisher","first-page":"1088","DOI":"10.3390\/ijms18051088","volume":"18","author":"Kwang-Youn Kim","year":"2017","unstructured":"Kim, K. Y. et al. Inhibition of Autophagy Promotes Salinomycin-Induced Apoptosis via Reactive Oxygen Species-Mediated PI3K\/AKT\/mTOR and ERK\/p38 MAPK-Dependent Signaling in Human Prostate Cancer Cells. Int J Mol Sci 18 (2017).","journal-title":"International Journal of Molecular Sciences"},{"key":"35815_CR44","first-page":"6020","volume":"61","author":"D Fukumura","year":"2001","unstructured":"Fukumura, D. et al. Hypoxia and acidosis independently up-regulate vascular endothelial growth factor transcription in brain tumors in vivo. Cancer Res 61, 6020\u20136024 (2001).","journal-title":"Cancer Res"},{"key":"35815_CR45","doi-asserted-by":"publisher","first-page":"828","DOI":"10.1096\/fj.05-5168hyp","volume":"20","author":"M Lopez-Lazaro","year":"2006","unstructured":"Lopez-Lazaro, M. HIF-1: hypoxia-inducible factor or dysoxia-inducible factor? Faseb J 20, 828\u2013832 (2006).","journal-title":"Faseb J"},{"key":"35815_CR46","doi-asserted-by":"publisher","first-page":"335","DOI":"10.1016\/j.ccr.2007.02.006","volume":"11","author":"JD Gordan","year":"2007","unstructured":"Gordan, J. D., Bertout, J. A., Hu, C. J., Diehl, J. A. & Simon, M. C. HIF-2alpha promotes hypoxic cell proliferation by enhancing c-myc transcriptional activity. Cancer Cell 11, 335\u2013347 (2007).","journal-title":"Cancer Cell"},{"key":"35815_CR47","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1186\/2049-3002-2-3","volume":"2","author":"N Masson","year":"2014","unstructured":"Masson, N. & Ratcliffe, P. J. Hypoxia signaling pathways in cancer metabolism: the importance of co-selecting interconnected physiological pathways. Cancer Metab 2, 3 (2014).","journal-title":"Cancer Metab"},{"key":"35815_CR48","first-page":"4610","volume":"60","author":"L Xu","year":"2000","unstructured":"Xu, L. & Fidler, I. J. Acidic pH-induced elevation in interleukin 8 expression by human ovarian carcinoma cells. Cancer Res 60, 4610\u20134616 (2000).","journal-title":"Cancer Res"},{"key":"35815_CR49","doi-asserted-by":"publisher","first-page":"6699","DOI":"10.1158\/0008-5472.CAN-06-0983","volume":"66","author":"EK Rofstad","year":"2006","unstructured":"Rofstad, E. K., Mathiesen, B., Kindem, K. & Galappathi, K. Acidic extracellular pH promotes experimental metastasis of human melanoma cells in athymic nude mice. Cancer Res 66, 6699\u20136707 (2006).","journal-title":"Cancer Res"},{"key":"35815_CR50","doi-asserted-by":"publisher","first-page":"11368","DOI":"10.1074\/jbc.M108347200","volume":"277","author":"L Xu","year":"2002","unstructured":"Xu, L., Fukumura, D. & Jain, R. K. Acidic extracellular pH induces vascular endothelial growth factor (VEGF) in human glioblastoma cells via ERK1\/2 MAPK signaling pathway: mechanism of low pH-induced VEGF. J Biol Chem 277, 11368\u201311374 (2002).","journal-title":"J Biol Chem"},{"key":"35815_CR51","doi-asserted-by":"publisher","first-page":"2032","DOI":"10.1021\/mp200292c","volume":"8","author":"JW Wojtkowiak","year":"2011","unstructured":"Wojtkowiak, J. W., Verduzco, D., Schramm, K. J. & Gillies, R. J. Drug resistance and cellular adaptation to tumor acidic pH microenvironment. Mol Pharm 8, 2032\u20132038 (2011).","journal-title":"Mol Pharm"},{"key":"35815_CR52","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1016\/j.semcancer.2017.01.007","volume":"43","author":"SK Parks","year":"2017","unstructured":"Parks, S. K. & Pouyssegur, J. Targeting pH regulating proteins for cancer therapy-Progress and limitations. Semin Cancer Biol 43, 66\u201373 (2017).","journal-title":"Semin Cancer Biol"},{"key":"35815_CR53","doi-asserted-by":"publisher","first-page":"3495","DOI":"10.1113\/jphysiol.2011.209072","volume":"589","author":"VA Skeberdis","year":"2011","unstructured":"Skeberdis, V. A., Rimkut\u0117, L., Skeberdyt\u0117, A., Paulauskas, N. & Bukauskas, F. F. pH-dependent modulation of connexin-based gap junctional uncouplers. J. Physiol. 589, 3495\u20133506 (2011).","journal-title":"J. Physiol."},{"key":"35815_CR54","doi-asserted-by":"crossref","first-page":"10350","DOI":"10.18632\/oncotarget.3585","volume":"6","author":"Y Zhou","year":"2015","unstructured":"Zhou, Y. et al. Salinomycin decreases doxorubicin resistance in hepatocellular carcinoma cells by inhibiting the beta-catenin\/TCF complex association via FOXO3a activation. Oncotarget 6, 10350\u201310365 (2015).","journal-title":"Oncotarget"},{"key":"35815_CR55","doi-asserted-by":"publisher","first-page":"62240","DOI":"10.18632\/oncotarget.11404","volume":"7","author":"N Mirkheshti","year":"2016","unstructured":"Mirkheshti, N. et al. Dual targeting of androgen receptor and mTORC1 by salinomycin in prostate cancer. Oncotarget 7, 62240\u201362254 (2016).","journal-title":"Oncotarget"},{"key":"35815_CR56","doi-asserted-by":"publisher","first-page":"305","DOI":"10.4161\/cbt.27309","volume":"15","author":"L Booth","year":"2014","unstructured":"Booth, L. et al. HDAC inhibitors enhance the lethality of low dose salinomycin in parental and stem-like GBM cells. Cancer Biol Ther 15, 305\u2013316 (2014).","journal-title":"Cancer Biol Ther"},{"key":"35815_CR57","doi-asserted-by":"publisher","first-page":"12877","DOI":"10.18632\/oncotarget.2657","volume":"5","author":"Z Xiao","year":"2014","unstructured":"Xiao, Z., Sperl, B., Ullrich, A. & Knyazev, P. Metformin and salinomycin as the best combination for the eradication of NSCLC monolayer cells and their alveospheres (cancer stem cells) irrespective of EGFR, KRAS, EML4\/ALK and LKB1 status. Oncotarget 5, 12877\u201312890 (2014).","journal-title":"Oncotarget"},{"issue":"Suppl 4","key":"35815_CR58","doi-asserted-by":"publisher","first-page":"989","DOI":"10.1289\/ehp.98106s4989","volume":"106","author":"PW Stacpoole","year":"1998","unstructured":"Stacpoole, P. W., Henderson, G. N., Yan, Z. & James, M. O. Clinical pharmacology and toxicology of dichloroacetate. Environ Health Perspect 106(Suppl 4), 989\u2013994 (1998).","journal-title":"Environ Health Perspect"},{"key":"35815_CR59","doi-asserted-by":"publisher","first-page":"1638","DOI":"10.1038\/onc.2012.198","volume":"32","author":"G Sutendra","year":"2013","unstructured":"Sutendra, G. et al. Mitochondrial activation by inhibition of PDKII suppresses HIF1a signaling and angiogenesis in cancer. Oncogene 32, 1638\u20131650 (2013).","journal-title":"Oncogene"},{"key":"35815_CR60","doi-asserted-by":"publisher","first-page":"4026","DOI":"10.1038\/onc.2011.113","volume":"30","author":"E Babu","year":"2011","unstructured":"Babu, E. et al. Role of SLC5A8, a plasma membrane transporter and a tumor suppressor, in the antitumor activity of dichloroacetate. Oncogene 30, 4026\u20134037 (2011).","journal-title":"Oncogene"},{"issue":"10","key":"35815_CR61","doi-asserted-by":"publisher","first-page":"1773","DOI":"10.1007\/s11605-008-0573-0","volume":"12","author":"Muthusamy Thangaraju","year":"2008","unstructured":"Thangaraju, M. et al. Sodium-coupled transport of the short chain fatty acid butyrate by SLC5A8 and its relevance to colon cancer. J Gastrointest Surg 12, 1773\u20131781; discussion 1781\u20131772 (2008).","journal-title":"Journal of Gastrointestinal Surgery"},{"key":"35815_CR62","doi-asserted-by":"crossref","first-page":"22414","DOI":"10.18632\/oncotarget.5628","volume":"8","author":"ZZ Zou","year":"2017","unstructured":"Zou, Z. Z. et al. Synergistic induction of apoptosis by salinomycin and gefitinib through lysosomal and mitochondrial dependent pathway overcomes gefitinib resistance in colorectal cancer. Oncotarget 8, 22414\u201322432 (2017).","journal-title":"Oncotarget"},{"key":"35815_CR63","doi-asserted-by":"publisher","first-page":"1831","DOI":"10.2217\/nnm-2016-0125","volume":"11","author":"F Xie","year":"2016","unstructured":"Xie, F. et al. Codelivery of salinomycin and chloroquine by liposomes enables synergistic antitumor activity in vitro. Nanomedicine (Lond) 11, 1831\u20131846 (2016).","journal-title":"Nanomedicine (Lond)"},{"key":"35815_CR64","doi-asserted-by":"publisher","first-page":"e97414","DOI":"10.1371\/journal.pone.0097414","volume":"9","author":"F Wang","year":"2014","unstructured":"Wang, F. et al. The synergistic in vitro and in vivo antitumor effect of combination therapy with salinomycin and 5-fluorouracil against hepatocellular carcinoma. PloS one 9, e97414 (2014).","journal-title":"PloS one"},{"key":"35815_CR65","doi-asserted-by":"publisher","first-page":"740564","DOI":"10.1155\/2011\/740564","volume":"2011","author":"J Tong","year":"2011","unstructured":"Tong, J. et al. Synergistic antitumor effect of dichloroacetate in combination with 5-fluorouracil in colorectal cancer. J Biomed Biotechnol 2011, 740564 (2011).","journal-title":"J Biomed Biotechnol"},{"key":"35815_CR66","doi-asserted-by":"publisher","first-page":"1246","DOI":"10.1007\/s10495-017-1394-y","volume":"22","author":"J Dewangan","year":"2017","unstructured":"Dewangan, J. et al. Novel combination of salinomycin and resveratrol synergistically enhances the anti-proliferative and pro-apoptotic effects on human breast cancer cells. Apoptosis 22, 1246\u20131259 (2017).","journal-title":"Apoptosis"},{"key":"35815_CR67","doi-asserted-by":"crossref","first-page":"59458","DOI":"10.18632\/oncotarget.10694","volume":"7","author":"B Li","year":"2016","unstructured":"Li, B. et al. Dichloroacetate and metformin synergistically suppress the growth of ovarian cancer cells. Oncotarget 7, 59458\u201359470 (2016).","journal-title":"Oncotarget"},{"key":"35815_CR68","doi-asserted-by":"publisher","first-page":"726","DOI":"10.3892\/ol.2012.552","volume":"3","author":"T Ishiguro","year":"2012","unstructured":"Ishiguro, T., Ishiguro, M., Ishiguro, R. & Iwai, S. Cotreatment with dichloroacetate and omeprazole exhibits a synergistic antiproliferative effect on malignant tumors. Oncol Lett 3, 726\u2013728 (2012).","journal-title":"Oncol Lett"},{"key":"35815_CR69","doi-asserted-by":"publisher","first-page":"2935","DOI":"10.3892\/mmr.2014.3080","volume":"11","author":"Q Xie","year":"2015","unstructured":"Xie, Q. et al. Combination of Taxol(R) and dichloroacetate results in synergistically inhibitory effects on Taxol-resistant oral cancer cells under hypoxia. Mol Med Rep 11, 2935\u20132940 (2015).","journal-title":"Mol Med Rep"},{"key":"35815_CR70","first-page":"11044","volume":"9","author":"S Zeng","year":"2016","unstructured":"Zeng, S., Liang, H. & Guan, G. Dichloroacetate enhances the cytotoxic effect of Cisplatin via decreasing the level of FOXM1 in prostate cancer. Int J Clin Med 9, 11044\u201311050 (2016).","journal-title":"Int J Clin Med"},{"key":"35815_CR71","doi-asserted-by":"publisher","first-page":"1499","DOI":"10.1159\/000369710","volume":"35","author":"XX Stander","year":"2015","unstructured":"Stander, X. X., Stander, B. A. & Joubert, A. M. Synergistic anticancer potential of dichloroacetate and estradiol analogue exerting their effect via ROS-JNK-Bcl-2-mediated signalling pathways. Cell Physiol Biochem 35, 1499\u20131526 (2015).","journal-title":"Cell Physiol Biochem"},{"key":"35815_CR72","doi-asserted-by":"publisher","first-page":"980","DOI":"10.1177\/1087057109341766","volume":"14","author":"F Hirschhaeuser","year":"2009","unstructured":"Hirschhaeuser, F., Leidig, T., Rodday, B., Lindemann, C. & Mueller-Klieser, W. Test system for trifunctional antibodies in 3D MCTS culture. J Biomol Screen 14, 980\u2013990 (2009).","journal-title":"J Biomol Screen"},{"key":"35815_CR73","doi-asserted-by":"publisher","first-page":"25418","DOI":"10.1074\/jbc.M114.547844","volume":"289","author":"A Hulikova","year":"2014","unstructured":"Hulikova, A., Aveyard, N., Harris, A. L., Vaughan-Jones, R. D. & Swietach, P. Intracellular carbonic anhydrase activity sensitizes cancer cell pH signaling to dynamic changes in CO2 partial pressure. J Biol Chem 289, 25418\u201325430 (2014).","journal-title":"J Biol Chem"},{"key":"35815_CR74","doi-asserted-by":"publisher","first-page":"1045","DOI":"10.1161\/01.RES.0000264071.11619.47","volume":"100","author":"P Swietach","year":"2007","unstructured":"Swietach, P., Rossini, A., Spitzer, K. W. & Vaughan-Jones, R. D. H+ ion activation and inactivation of the ventricular gap junction: a basis for spatial regulation of intracellular pH. Circ Res 100, 1045\u20131054 (2007).","journal-title":"Circ Res"},{"key":"35815_CR75","doi-asserted-by":"publisher","first-page":"3744","DOI":"10.1158\/0008-5472.CAN-15-1862","volume":"76","author":"A McIntyre","year":"2016","unstructured":"McIntyre, A. et al. Disrupting Hypoxia-Induced Bicarbonate Transport Acidifies Tumor Cells and Suppresses Tumor Growth. Cancer Res 76, 3744\u20133755 (2016).","journal-title":"Cancer Res"},{"key":"35815_CR76","doi-asserted-by":"publisher","first-page":"309","DOI":"10.1038\/nprot.2008.226","volume":"4","author":"J Friedrich","year":"2009","unstructured":"Friedrich, J., Seidel, C., Ebner, R. & Kunz-Schughart, L. A. Spheroid-based drug screen: considerations and practical approach. Nat Protoc 4, 309\u2013324 (2009).","journal-title":"Nat Protoc"},{"key":"35815_CR77","doi-asserted-by":"crossref","unstructured":"Chen, W. et al. High-throughput image analysis of tumor spheroids: a user-friendly software application to measure the size of spheroids automatically and accurately. J Vis Exp (2014).","DOI":"10.3791\/51639"},{"key":"35815_CR78","doi-asserted-by":"publisher","first-page":"27","DOI":"10.2174\/1567201043480036","volume":"1","author":"H Glavinas","year":"2004","unstructured":"Glavinas, H., Krajcsi, P., Cserepes, J. & Sarkadi, B. The role of ABC transporters in drug resistance, metabolism and toxicity. Curr Drug Deliv 1, 27\u201342 (2004).","journal-title":"Curr Drug Deliv"},{"key":"35815_CR79","doi-asserted-by":"publisher","first-page":"402","DOI":"10.1006\/meth.2001.1262","volume":"25","author":"KJ Livak","year":"2001","unstructured":"Livak, K. J. & Schmittgen, T. D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25, 402\u2013408 (2001).","journal-title":"Methods"},{"key":"35815_CR80","doi-asserted-by":"publisher","first-page":"765","DOI":"10.1093\/bioinformatics\/btp053","volume":"25","author":"DJ McCarthy","year":"2009","unstructured":"McCarthy, D. J. & Smyth, G. K. Testing significance relative to a fold-change threshold is a TREAT. Bioinformatics 25, 765\u2013771 (2009).","journal-title":"Bioinformatics"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-35815-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-35815-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-35815-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T12:24:55Z","timestamp":1671625495000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-35815-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,12,10]]},"references-count":80,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2018,12]]}},"alternative-id":["35815"],"URL":"https:\/\/doi.org\/10.1038\/s41598-018-35815-4","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,12,10]]},"assertion":[{"value":"11 May 2018","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"7 November 2018","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 December 2018","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"17744"}}