Abstract
Although a wide variety of cytotoxic plant extracts and phytochemicals are known to act synergistically with anticancer drug doxorubicin (D), their clinical application is hindered by safety concerns of such combination therapy. Our earlier studies showed that red beetroot (Beta vulgaris L.) extract (B), approved by Food and Drug Administration and European Union as red food color E162, reduced multi-organ tumor formations in various animal models when administered in drinking water. This led us to postulate that a long-term daily exposure to low doses of B through diet might be safe and sufficient to produce cancer chemopreventive effect in humans. Further, our recent comparative cytotoxic investigation with B and D in several human cancer cell lines indicated their potential for synergistic activity. Since B is considered safe for human use with no known toxicity, we conducted the present study to evaluate its synergistic antiproliferative activity with D against pancreatic (PaCa), breast (MCF-7) and prostate (PC-3) tumor cells of human origin. Different concentrations of B and D (0.29–290 μg/ml) and in various combinations (B:D ratio = 1:0, 1:1, 5:1, 1:5 and 0:1) were tested for cytotoxic effects against the three cancer cells. The viability of cells was assessed after 72 h incubation with various combinations of B and D using the trypan-blue staining method. The cytotoxic data were analyzed by the combination index method of Chou and Talalay to establish synergy between B and D. The results indicated that an overall positive reduction in drug concentration was achieved by D when combined with B in its cytotoxicity profile in the three human cancer cells tested. The synergistic cytotoxicity was best when the B:D ratio of 1:5 was used in PaCa cells at IC50, IC75 and IC90 dose levels and in MCF-7 cells at IC90 dose level. These results warrant further studies on the potential of red beetroot extract–doxorubicin combination in treating human cancers.
References
1. Government of India, Ministry of Health and Family Welfare. The Ayurvedic Pharmacopoeia of India, Parts I and II. New Delhi: Government of India Press, 1978 and 2007.Search in Google Scholar
2. Bensky D, Gamble A. Chinese herbal medicine: materia medica, rev. ed. Seattle, WA: Eastland Press, 1993.Search in Google Scholar
3. Patwardhan B, Warude D, Pushpnagadan P, Bhat N. Ayurveda and traditional Chinese medicine: a comparative overview. Evid Based Complement Alternat Med 2005;2:465–73.10.1093/ecam/neh140Search in Google Scholar
4. Chou TC. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism of drug combination studies. Pharmacol Rev 2006;58:621–81.10.1124/pr.58.3.10Search in Google Scholar
5. Chou TC. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res 2010;70:440–6.10.1158/0008-5472.CAN-09-1947Search in Google Scholar
6. Ulrich-Merzenich G, Panek D, Zeitler H, Vetter H, Wagner H. Drug development from natural products: exploiting synergistic effects. Indian J Exp Biol 2010;48:208–19.Search in Google Scholar
7. Tanos V, Brzezinski A, Drize O, Strauss N, Peretz T. Synergistic inhibitory effects of genistein and tamoxifen on human dysplastic and malignant epithelial breast cells in vitro. Eur J Obstet Gynecol Reprod Biol 2002;102:188–94.10.1016/S0301-2115(01)00582-6Search in Google Scholar
8. Wang MY, West BJ, Jensen CJ, Nowicki D, Chen SU, Palu AK, et al. Morinda citrifolia (Noni): A literature review and recent advances in Noni research. Acta Pharmacol Sin 2002:12:1127–47.Search in Google Scholar
9. Sharma G, Tyagi AK, Singh RP, Chan DC, Agarwal R. Synergistic anticancer effects of grape seed extract and conventional cytotoxic agent doxorubicin against human breast carcinoma cells. Breast Cancer Res Treat 2004;85:1–12.10.1023/B:BREA.0000020991.55659.59Search in Google Scholar
10. Tyagi AK, Agarwal C, Chan DC, Agarwal R. Synergistic anticancer effects of silibinin with conventional cytotoxic agents doxorubicin, cisplatin and carboplatin against human breast carcinoma MCF-7 and MDA-MB468 cells. Oncol Res 2004;11:493–9.Search in Google Scholar
11. Hemaiswarya S, Doble M. Potential synergism of natural products in the treatment of cancer. Phytother Res 2006;20:239–49.10.1002/ptr.1841Search in Google Scholar PubMed
12. Wang CZ, Luo X, Zhang B, Song WX, Ni M, Mehendale S, et al. Notoginseng enhances anti-cancer effects of 5-fluorouracil on human colorectal cancer cells. Cancer Chemother Pharmacol 2007;60:69–79.10.1007/s00280-006-0350-2Search in Google Scholar PubMed PubMed Central
13. Wei J, Liu B, Wang L, Qian X, Ding Y, Yu L. Synergistic interaction between tetrandrine and chemotherapeutic agents and influence of tetrandrine on therapeutic agent-associated genes in human gastric cancer cell lines. Cancer Chemother Pharmacol 2007;60:703–11.10.1007/s00280-007-0416-9Search in Google Scholar PubMed
14. Akao Y, Nakagawa Y, Inuma M, Nozawa Y. Anti-cancer effects of xanthones from pericarps of mangosteen: Int J Mol Sci 2008;9:355–70.10.3390/ijms9030355Search in Google Scholar PubMed PubMed Central
15. Banerjee S, Kaseb AO, Wang Z, Kong D, Mohammad M, Padhye S, et al. Antitumor activity of gemcitabine and oxaliplatin is augmented by thymoquinone in pancreatic cells. Cancer Res 2009;69:5575–83.10.1158/0008-5472.CAN-08-4235Search in Google Scholar PubMed
16. Griggs JJ. Reducing the toxicity of anticancer therapy: new strategies. Leuk Res 1998;22(Suppl 1):S27–33.10.1016/S0145-2126(98)00036-8Search in Google Scholar
17. Borek C. Dietary antioxidants and human cancer. Integr Cancer Ther 2004;3:333–41.10.1177/1534735404270578Search in Google Scholar
18. Liu HR. Potential synergy of phytochemicals in cancer prevention: mechanisms of action. J Nutr 2004;134:3479S–85S.10.1093/jn/134.12.3479SSearch in Google Scholar
19. Huang XZ, Wang J, Huang C, Chen YY, Shi GY, Hu QS, Yi J. Emodin enhances cytotoxicity of chemotherapeutic drugs in prostate cancer cells: the mechanisms involve ROS-mediated suppression of multidrug resistance and hypoxia inducible factor-1. Cancer Biol Ther 2008;7:468–75.10.4161/cbt.7.3.5457Search in Google Scholar
20. Chai S, To KKW, Lin G. Circumvention of multi-drug resistance of cancer cells by Chinese herb medicine. Chin Med 2010;5:26–35.10.1186/1749-8546-5-26Search in Google Scholar
21. Tan W, Lu J, Huang M, Li Y, Chen M, Wu G, et al. Review: anti-cancer natural products isolated from Chinese medicinal herbs. Chin Med 2011;6:27–42.10.1186/1749-8546-6-27Search in Google Scholar
22. Lown JW. Anthracycline and anthraquinone anticancer agents. Current status and recent developments. Pharmacol Ther 1993;60:185–214.10.1016/0163-7258(93)90006-YSearch in Google Scholar
23. Carvalho C, Santos RX, Cardoso S, Correia S, Oliveira PJ, Santos MS, Moreira PI. Doxorubicin: the good, the bad and the ugly effect. Curr Med Chem 2009;16:3267–85.10.2174/092986709788803312Search in Google Scholar
24. Abdel-Wahab MH, El-Mahdy MA, Abd-Ellah MF, Helat GK, Khalifa F, Hamada FMA. Influence of p-coumaric acid on doxorubicin-induced oxidative stress in rat’s heart. Pharmacol Res 2003;48:461–5.10.1016/S1043-6618(03)00214-7Search in Google Scholar
25. Psotova J, Chlopcikova S, Miketova P, Hrbac J, Simanek V. Chemoprotective effect of plant phenolics against anthracycline-induced toxicity on rat cardiomyocytes. Part III. Apigenin, baicalenin, kaempherol, luteolin and quercetin. Phytother Res 2004;18:516–21.Search in Google Scholar
26. Teiten MH, Eifes S, Dicato M, Diedrich M. Review: Curcumin – the paradigm of a multi-target natural compound with applications in cancer prevention and treatment. Toxins 2010;2:128–62.10.3390/toxins2010128Search in Google Scholar PubMed PubMed Central
27. Ravizza R, Gariboldi MB, Molteni R, Monti E. Linalool, a plant-derived monoterpene alcohol, reverses doxorubicin resistance in human breast adenocarcinoma cells. Oncol Rep 2008;20:625–30.Search in Google Scholar
28. Effenberger K, Schobert R. Combinational effects of thymoquinone on the anti-cancer activity of doxorubicin. Cancer Chemother Pharmacol 2010;67:867–74.10.1007/s00280-010-1386-xSearch in Google Scholar PubMed
29. Kaneez FS, Al-Salam S, Alaaeldin AH. Sesame oil as a protective agent against doxorubicin induced cardiotoxicity in rat. Am J Pharmacol Toxicol 2007;2:159–63.10.3844/ajptsp.2007.159.163Search in Google Scholar
30. Alkreathy H, Damanhouri ZA, Ahmed N, Slevin M, Ali SS and Osman AMM. Aged garlic extract protects against doxorubicin-induced cardiotoxicity in rats. Food Chem Toxicol 2010;48:951–6.10.1016/j.fct.2010.01.005Search in Google Scholar PubMed
31. Liu TJ, Yeh YC, Ting CT, Lee WL, Wang LC, Lee HW, et al. Ginkgo biloba extract 761 reduces doxorubicin-induced apoptoic damage in rat hearts and neonatal cardiomyocytes. Cardiovasc Res 2008;80:227–35.10.1093/cvr/cvn192Search in Google Scholar PubMed
32. Wattanapitayakul SK, Chularojmontri L, Herunsalee A, Charuchongkolwongse S, Niumsakul S, Bauer JA. Screening of antioxidants from medicinal plants for cardioprotective effect against doxorubicin toxicity. Basic Clin Pharmacol Toxicol 2005;96:80–7.10.1111/j.1742-7843.2005.pto960112.xSearch in Google Scholar PubMed
33. Breibart E. Effects of water-soluble antioxidants from spinach, NAO, on doxorubicin-induced heart injury. Hum Exp Toxicol 2001;20:337–45.10.1191/096032701680350604Search in Google Scholar PubMed
34. Liang G, Tang A, Lin X, Li L, Zhang S, Huang Z, et al. Green tea catechins augment the antitumor activity of doxorubicin in an in vivo mouse model for chemoresistant liver cancer. Int J Oncol 2010;37:111–23.Search in Google Scholar
35. Sarkar A, Sreenivasan Y, Ramesh GT, Manna SK. D-Glucoside suppresses tumor necrosis factor-induced activation of nuclear transcription factor kB but potentiates apoptosis. J Biol Chem 2004;279:33768–81.10.1074/jbc.M403424200Search in Google Scholar PubMed
36. Kolarovic J, Popovic M, Zlinska J, Trivic S, Vojnovic M. Antioxidant activities of celery and parsley juices in rats treated with doxorubicin. Molecules 2010;15:6193–204.10.3390/molecules15096193Search in Google Scholar PubMed PubMed Central
37. Nair PKR, Melnick S, Wnuk SF, Rapp M, Escalon E, Ramachandran C. Isolation and Characterization of an anticancer catechol compound from Semecarpus anacradium. J Ethnopharmacol 2009;122:450–6.10.1016/j.jep.2009.02.001Search in Google Scholar PubMed
38. Colombo V, Lupi M, Falcetta F, Forestieri D, D’Incalci M, Ubezio P. Chemotherapeutic activity of silymarin combined with doxorubicin or paclitaxel in sensitive and multidrug-resistant colon cancer cells. Cancer Chemother Pharmacol 2011;67:369–79.10.1007/s00280-010-1335-8Search in Google Scholar PubMed
39. Chen HM, Wu YC, Chia YC, Chang FR, Hsu HK, Hseih YC, et al. Gallic acid, a major component of Toona sinensis leaf extracts, contains ROS-mediated anti-cancer activity in human prostate cancer cells. Cancer Lett 2009;286:161–71.10.1016/j.canlet.2009.05.040Search in Google Scholar
40. Pandey MK, Sung B, Ahn KS, Kunnumakkara AB, Chaturvedi MM, Aggarwal BB. Gambogic acid, a new ligand for transferring receptor, potentiates TNF-induced apoptosis through modulation of the nuclear factor-kB signaling pathway. Blood 2007;110:3517–25.10.1182/blood-2007-03-079616Search in Google Scholar
41. Scambia G, Ranelletti FO, Panici PB, De Vencenzo R, Bonanno G, Piantelli M, et al. Quercetin potentiates the effect of adriamycin in a multi-drug resistant MCF-7 human breast cancer line: P- glycoprotein as a possible target. Cancer Chemother Pharmacol 1994:34:459–64. Comment by Sarkar MA. Quercetin not only inhibits P-glycoprotein efflux but also inhibits CYP3A isoenzyme. Cancer Chemother Pharmacol 1995:36:448–50.10.1007/BF00685655Search in Google Scholar
42. Sun AX, Ye XG, Li CY, Xue BY, Li LF, Cao XF, et al. Synergistic anticancer effects of tetrandrine combined with doxorubicin or vincristine in vitro. Acta Pharmacol Sin 1999;20:69–73.Search in Google Scholar
43. Chanamahasathein W, Ampasavate C, Greger H, Limtrakul P. Stemona alkaloids, from traditional Thai medicine, increase chemosensitivity via P-glycoprotein-mediated multidrug resistance. Phytomedicine 2011;18:199–204.10.1016/j.phymed.2010.07.014Search in Google Scholar
44. Fenig E, Nordenberg J, Beery E, Sulkes J, Wasserman L. Combined effect of aloe-emodin and chemotherapeutic agents on the proliferation of an adherent variant cell line of Merkel cell carcinoma. Oncol Rep 2004;11:213–7.10.3892/or.11.1.213Search in Google Scholar
45. Gao RL, Lin XJ, Qian XD, Chen XH, Nu YP. Effect of ginsenosides and panaxatriol extracted from ginseng on inhibition of proliferation, inducing apoptosis and cytotoxic drug sensitivity in leukemic cells. J Chin Med Res 2004;4:97–9.Search in Google Scholar
46. Zheng Y, Zhou F, Wu X, Wen X, Li Y, Yan B, et al. 23-Hydroxybetulinic acid from Pulsatilla chinensis (Bunge) Regel synergizes the antitumor activities of doxorubicin in vitro and in vivo. J Ethnopharmacol 2010;128:615–22.10.1016/j.jep.2010.02.004Search in Google Scholar
47. Pinmai K, Chunlaratthanabhorn S, Ngamkitidechakul C, Soonthornchareon N, Hahnvajanawong C. Synergistic growth inhibitory effects of Phyllanthus emblica and Terminalia bellerica extracts with conventional cytotoxic agents: Doxorubicin and cisplatin against human hepatocelluar carcinoma and lung cancer cells. World J Gastroenterol 2008;14:1491–710.3748/wjg.14.1491Search in Google Scholar
48. Francis FJ. Safety of food colorants. In: Hendry GAF, Hougton JD, editors. Natural food colorants, 2nd ed. Glasgow: Blackie Academic Professional, 1996:112–30.Search in Google Scholar
49. Kapadia GJ, Azuine MA, Rao GS, Arai T, Iida A, Tokuda H. Cytotoxic effect of the red beetroot (Beta vulgaris L.) extract compared to doxorubicin (Adriamycin) in the human prostate (PC-3) and breast (MCF-7) cancer cell lines. Anti-Cancer Agent Med Chem 2011;11:280–4.10.2174/187152011795347504Search in Google Scholar
50. Kapadia GJ, Rao GS, Elinor-Reid T, Iida A, Suzuki N, Tokuda H. Synergistic cytotoxic effect of red beetroot (Beta vulgaris L.) food colorant E162 with anticancer drug doxorubicin (Adriamycin) against human prostate, breast and pancreatic tumor cells. In: Abstracts of Papers, American Society of Pharmacognosy 52nd Annual Meeting, San Diego, CA, 2011:P241.Search in Google Scholar
51. Kapadia GJ, Tokuda H, Konoshima T, Nishino H. Chemoprevention of lung and skin cancer by Beta vulgaris (beet) root extract. Cancer Lett 1996;100:211–4.10.1016/0304-3835(95)04087-0Search in Google Scholar
52. Kapadia GJ, Azuine MA, Sridhar R, Okuda Y, Tsuruta A, Ichishi E, et al. Chemoprevention of DMBA-induced UV-B promoted, NOR-1-induced TPA promoted skin carcinogenesis and DEN-induced phenobarbital promoted liver tumors in mice by extract of beetroot. Pharmacol Res 2003;47:141–8.10.1016/S1043-6618(02)00285-2Search in Google Scholar
53. Kapadia GJ, Rao GS. Anticancer effects of red beet pigments. In: Neelwarne B, editor. Red beet biotechnology: metabolites for food and pharmaceutical applications. New York, NY: Springer 2013:125–54.10.1007/978-1-4614-3458-0_7Search in Google Scholar
54. Chou TC, Talalay P. Analysis of combined drug effects: a new look at a very old problem. Trend Pharmacol Sci 1983;4:450–4.10.1016/0165-6147(83)90490-XSearch in Google Scholar
55. Reznikoff CA, Bertram JS, Brankow DW, Heidelberger C. Quantitative and qualitative studies of chemical transformation of cloned C3H mouse embryo cells sensitive to post confluence inhibition of cell division. Cancer Res 1973;33:3239–49.Search in Google Scholar
56. Kakunga TA. A quantitative system for assay of malignant trans- formation by chemical carcinogens using a clone of BALB-373. Int J Cancer 1973;12:463–73.10.1002/ijc.2910120217Search in Google Scholar PubMed
57. Tolnai S. A method for viable cell count. Method Cell Sci 1975;1:37–8.10.1007/BF00914435Search in Google Scholar
58. Azeredo HMC. Betalains: properties, sources, applications, and stability – a review. Int J Food Sci Technol 2009;44:2365–76.10.1111/j.1365-2621.2007.01668.xSearch in Google Scholar
59. Ferreira ALA, Yeum KJ, Matsubara LS, Matsubara BB, Correa CR, Periera EJ, et al. Doxorubicin as an antioxidant: maintenance of myocardial levels of lycopene under doxorubicin treatment. Free Radic Biol Med 2007;43:740–51.10.1016/j.freeradbiomed.2007.05.002Search in Google Scholar PubMed
©2013 by Walter de Gruyter Berlin / Boston
