Abstract
Cisplatin (cDDP) is one of the most widely used anticancer-drugs in both therapy and research. However, cDDP-resistance is the greatest obstacle for the successful treatment of cancer patients. In the present study, the possible joint anticancer effect of bee venom (BV), as a natural toxin, and cDDP towards human glioblastoma A1235 cells was evaluated. Treatment with BV alone in concentrations of 2.5–30 μg/ml displayed dose-dependent cytotoxicity towards A1235 cells, as evaluated with different cytotoxicity assays (MTT, Cristal violet and Trypan blue exclusion assay), with an IC50 value of 22.57 μg/ml based on the MTT results. Furthermore, BV treatment induced necrosis, which was confirmed by typical morphological features and fast staining with ethidium-bromide dye. Pre-treatment with BV induced cell sensitization to cDDP, indicating that BV could improve the killing effect of selected cells when combined with cDDP. The isobologram method used to determine the extent of synergism in combining two agents to examine their possible therapeutic effect showed that combined treatment induced an additive and/or synergistic effect towards selected cells depending on the concentration of both. Hence, a greater anticancer effect could be triggered if BV was used in the course of chemotherapy. The obtained results indicate that joint treatment with BV could be useful from the point of minimizing the cDDP concentration during chemotherapy, thus reducing and/or postponing the development of drug resistance. Our data, in accordance with previously reported results, suggests that BV could be used in the development of a new strategy for cancer treatment.
References
Alizadehnohi M, Nabiuni M, Nazari Z, Safaeinejad Z, Irian S (2012) The synergistic cytotoxic effect of cisplatin and honey bee venom on human ovarian cancer cell line A2780cp. J Venom Res 3:22–27
Boukraâ L, Sulaiman SA (2009) Rediscovering the antibiotics of the hive. Recent Pat Antiinfect Drug Discov 4:206–213. doi:10.2174/157489109789318505
Brdar B, Matulić M (1988) Induction of plasminogen activator by N-methyl-N’-nitro-N-nitrosoguanidine in mer+ and mer− human tumour cell strains. Carcinogenesis 9:2191–2195. doi:10.1093/carcin/9.12.2191
Brozović A, Ambriović-Ristov A, Osmak M (2010) The relationship between cisplatin-induced reactive oxygen species, glutathione, and BCL-2 and resistance to cisplatin. Crit Rev Toxicol 40:347–359. doi:10.3109/10408441003601836
Carmona-Ribeiro AM, de Melo Carrasco LD (2014) Novel formulations for antimicrobial peptides. Int J Mol Sci 15:18040–18083. doi:10.3390/ijms151018040
Chen J, Lariviere WR (2010) The nociceptive and anti-nociceptive effects of bee venom injection and therapy: a double-edged sword. Prog Neurobiol 92:151–183. doi:10.1016/j.pneurobio.2010.06.006
Choi KE, Hwang CJ, Gu SM, Park MH, Kim JH, Park JH, Ahn YJ, Kim JY, Song MJ, Song HS, Han SB, Hong JT (2014) Cancer cell growth inhibitory effect of bee venom via increase of death receptor 3 expression and inactivation of NF-kappa B in NSCLC cells. Toxins (Basel) 6:2210–2228. doi:10.3390/toxins6082210
Dasari S, Tchounwou PB (2014) Cisplatin in cancer therapy: molecular mechanisms of action. Eur J Pharmacol 740:364–378. doi:10.1016/j.ejphar.2014.07.025
Dempsey CE (1990) The actions of melittin on membranes. Biochim Biophys Acta 1031:143–161. doi:10.1016/0304-4157(90)90006-X
Deorukhkar A, Krishnan S, Sethi G, Aggarwal BB (2007) Back to basics: how natural products can provide the basis for new therapeutics. Expert Opin Investig Drugs 16:1753–1773. doi:10.1517/13543784.16.11.1753
Edinger AL, Thompson CB (2004) Death by design: apoptosis, necrosis and autophagy. Curr Opin Cell Biol 16:663–669. doi:10.1016/j.ceb.2004.09.011
Fink D, Howell SB (2000) How does cisplatin kill cells? In: Kelland LR, Farrell N (eds) Platinum-based drugs in cancer therapy 7. Humana Press Inc, Totowa, New Jersey, pp 149–167
Fuertes MA, Alonso C, Perez JM (2003) Biochemical modulation of cisplatin mechanisms of action: enhancement of antitumor activity and circumvention of drug resistance. Chem Rev 103:645–662. doi:10.1021/cr020010d
Gajski G, Garaj-Vrhovac V (2011) Bee venom induced cytogenetic damage and decreased cell viability in human white blood cells after treatment in vitro: a multi-biomarker approach. Environ Toxicol Pharmacol 32:201–211. doi:10.1016/j.etap.2011.05.004
Gajski G, Garaj-Vrhovac V (2013) Melittin: a lytic peptide with anticancer properties. Environ Toxicol Pharmacol 36:697–705. doi:10.1016/j.etap.2013.06.009
Gajski G, Čimbora-Zovko T, Osmak M, Garaj-Vrhovac V (2011) Bee venom and melittin are cytotoxic against different types of tumor and non-tumor cell lines in vitro. Cancer Res J 4:159–174
Gajski G, Čimbora-Zovko T, Rak S, Rožman M, Osmak M, Garaj-Vrhovac V (2014) Combined antitumor effects of bee venom and cisplatin on human cervical and laryngeal carcinoma cells and their drug resistant sublines. J Appl Toxicol 34:1332–1341. doi:10.1002/jat.2959
Garaj-Vrhovac V, Gajski G (2009) Evaluation of the cytogenetic status of human lymphocytes after exposure to a high concentration of bee venom in vitro. Arh Hig Rada Toksikol 60:27–34. doi:10.2478/10004-1254-60-2009-1896
Gaspar D, Veiga AS, Castanho MA (2013) From antimicrobial to anticancer peptides: a review. Front Microbiol 4:294. doi:10.3389/fmicb.2013.00294
Golstein P, Kroemer G (2007) Cell death by necrosis: towards a molecular definition. Trends Biochem Sci 32:37–43. doi:10.1016/j.tibs.2006.11.001
Gotay CC (2010) Cancer prevention: major initiatives and looking into the future. Expert Rev Pharmacoecon Outcomes Res 10:143–154. doi:10.1586/erp.10.9
Hong SJ, Rim GS, Yang HI, Yin CS, Koh HG, Jang MH, Kim CJ, Choe BK, Chung JH (2005) Bee venom induces apoptosis through caspase-3 activation in synovial fibroblasts of patients with rheumatoid arthritis. Toxicon 46:39–45. doi:10.1016/j.toxicon.2005.03.015
Hu H, Chen D, Li Y, Zhang X (2006) Effect of polypeptides in bee venom on growth inhibition and apoptosis induction of the human hepatoma cell line SMMC-7721 in vitro and Balb/c nude mice in-vivo. J Pharm Pharmacol 58:83–89. doi:10.1211/jpp.58.1.0010
Ip SW, Liao SS, Lin SY, Lin JP, Yang JS, Lin ML, Chen GW, Lu HF, Lin MW, Han SM, Chung JG (2008a) The role of mitochondria in bee venom-induced apoptosis in human breast cancer MCF7 cells. In Vivo 22:237–245
Ip SW, Wei HC, Lin JP, Kuo HM, Liu KC, Hsu SC, Yang JS, Mei-Dueyang Chiu TH, Han SM, Chung JG (2008b) Bee venom induced cell cycle arrest and apoptosis in human cervical epidermoid carcinoma Ca Ski cells. Anticancer Res 28:833–842
Ip SW, Chu YL, Yu CS, Chen PY, Ho HC, Yang JS, Huang HY, Chueh FS, Lai TY, Chung JG (2012) Bee venom induces apoptosis through intracellular Ca(2+) -modulated intrinsic death pathway in human bladder cancer cells. Int J Urol 19:61–70. doi:10.1111/j.1442-2042.2011.02876.x
Jang MH, Shin MC, Lim S, Han SM, Park HJ, Shin I (2003) Bee venom induces apoptosis and inhibits expression of cyclooxygenase-2 mRNA in human lung cancer cell line NCI-H1299. J Pharmacol Sci 91:95–104. doi:10.1254/jphs.91.95
Jin ZJ (1980) Addition in drug combination. Acta Pharmacol Sin 1:70–76
Jo M, Park MH, Kollipara PS, An BJ, Song HS, Han SB, Kim JH, Song MJ, Hong JT (2012) Anti-cancer effect of bee venom toxin and melittin in ovarian cancer cells through induction of death receptors and inhibition of JAK2/STAT3 pathway. Toxicol Appl Pharmacol 258:72–81. doi:10.1016/j.taap.2011.10.009
Kouros N (2013) New research finds HIV can be killed with bee venom. Monash Bioeth Rev 31:4
Kueng W, Silber E, Eppenberger U (1989) Quantification of cells cultured on 96-well plates. Anal Biochem 182:16–19. doi:10.1016/0003-2697(89)90710-0
Lee YJ, Kang SJ, Kim BM, Kim YJ, Woo HD, Chung HW (2007) Cytotoxicity of honeybee (Apis mellifera) venom in normal human lymphocytes and HL-60 cells. Chem Biol Interact 169:189–197. doi:10.1016/j.cbi.2007.06.036
Liu X, Chen D, Xie L, Zhang R (2002) Effect of honey bee venom on proliferation of K1735M2 mouse melanoma cells in vitro and growth of murine B16 melanomas in-vivo. J Pharm Pharmacol 54:1083–1089. doi:10.1211/002235702320266235
Liu CC, Yang H, Zhang LL, Zhang Q, Chen B, Wang Y (2014) Biotoxins for cancer therapy. Asian Pac J Cancer Prev 15:4753–4758. doi:10.7314/APJCP.2014.15.12.4753
Macciò A, Madeddu C (2013) Cisplatin: an old drug with a newfound efficacy— from mechanisms of action to cytotoxicity. Expert Opin Pharmacother 14:1839–1857. doi:10.1517/14656566.2013.813934
Mickisch G, Fajta S, Keilhauer G, Schlick E, Tschada R, Alken P (1990) Chemosensitivity testing of primary human renal cell carcinoma by a tetrazolium based microculture assay (MTT). Urol Res 18:131–136. doi:10.1007/BF00302474
Moon DO, Park SY, Heo MS, Kim KC, Park C, Ko WS (2006) Key regulators in bee venom-induced apoptosis are Bcl-2 and caspase-3 in human leukemic U937 cells through downregulation of ERK and Akt. Int Immunopharmacol 6:1796–1807. doi:10.1016/j.intimp.2006.07.027
Oršolić N (2009) Potentiation of bleomycin lethality in HeLa and V79 cells by bee venom. Arh Hig Rada Toksikol 60:317–326. doi:10.2478/10004-1254-60-2009-1936
Oršolić N (2012) Bee venom in cancer therapy. Cancer Metastasis Rev 31:173–194. doi:10.1007/s10555-011-9339-3
Oršolić N, Šver L, Verstovšek S, Terzić S, Bašić I (2003) Inhibition of mammary carcinoma cell proliferation in vitro and tumor growth in vivo by bee venom. Toxicon 41:861–870. doi:10.1016/S0041-0101(03)00045-X
Park MH, Choi MS, Kwak DH, Oh KW, do Yoon Y, Han SB, Song HS, Song MJ, Hong JT (2011) Anti-cancer effect of bee venom in prostate cancer cells through activation of caspase pathway via inactivation of NF-κB. Prostate 71:801–812. doi:10.1002/pros.21296
Premratanachai P, Chanchao C (2014) Review of the anticancer activities of bee products. Asian Pac J Trop Biomed 4:337–344. doi:10.12980/APJTB.4.2014C1262
Putz T, Ramoner R, Gander H, Rahm A, Bartsch G, Thurnher M (2006) Antitumor action and immune activation through cooperation of bee venom secretory phospholipase A2 and phosphatidylinositol-(3,4)-bisphosphate. Cancer Immunol Immunother 55:1374–1383. doi:10.1007/s00262-006-0143-9
Raghuraman H, Chattopadhyay A (2007) Melittin: a membrane-active peptide with diverse functions. Biosci Rep 27:189–223. doi:10.1007/s10540-006-9030-z
Safaeinejad Z, Nabiuni M, Nazari Z (2013) Potentiation of a novel palladium (II) complex lethality with bee venom on the human T-cell acute lymphoblastic leukemia cell line (MOLT-4). J Venom Anim Toxins Incl Trop Dis 19:25. doi:10.1186/1678-9199-19-25
Schweizer F (2009) Cationic amphiphilic peptides with cancer-selective toxicity. Eur J Pharmacol 625:190–194. doi:10.1016/j.ejphar.2009.08.043
Shen DW, Pouliot LM, Hall MD, Gottesman MM (2012) Cisplatin resistance: a cellular self-defense mechanism resulting from multiple epigenetic and genetic changes. Pharmacol Rev 64:706–721. doi:10.1124/pr.111.005637
Six DA, Dennis EA (2000) The expanding superfamily of phospholipase A(2) enzymes: classification and characterization. Biochim Biophys Acta 1488:1–19. doi:10.1016/S1388-1981(00)00105-0
Son DJ, Lee JW, Lee YH, Song HS, Lee CK, Hong JT (2007) Therapeutic application of anti-arthritis, pain-releasing, and anti-cancer effects of bee venom and its constituent compounds. Pharmacol Ther 115:246–270. doi:10.1016/j.pharmthera.2007.04.004
Stathopoulos GP (2013) Cisplatin: process and future. J BUON 18:564–569
Stewart DJ (2007) Mechanisms of resistance to cisplatin and carboplatin. Crit Rev Oncol Hematol 63:12–31. doi:10.1016/j.critrevonc.2007.02.001
Stewart BW, Kleihues P (2003) World cancer report, 2nd edn. IARC Press, Lyon
Strober W (2001) Trypan blue exclusion test of cell viability. Curr Protoc Immunol A 3:A3B. doi:10.1002/0471142735.ima03bs21
Stuhlmeier KM (2007) Apis mellifera venom and melittin block neither NF-kappa B-p50-DNA interactions nor the activation of NF-kappa B, instead they activate the transcription of proinflammatory genes and the release of reactive oxygen intermediates. J Immunol 179:655–664. doi:10.4049/jimmunol.179.1.655
Tu WC, Wu CC, Hsieh HL, Chen CY, Hsu SL (2008) Honeybee venom induces calcium-dependent but caspase-independent apoptotic cell death in human melanoma A2058 cells. Toxicon 52:318–329. doi:10.1016/j.toxicon.2008.06.007
Vanden Berghe T, Linkermann A, Jouan-Lanhouet S, Walczak H, Vandenabeele P (2014) Regulated necrosis: the expanding network of non-apoptotic cell death pathways. Nat Rev Mol Cell Biol 15:135–147. doi:10.1038/nrm3737
Varanda EA, Tavares DC (1998) Radioprotection: mechanism and radioprotective agents including honey bee venom. J Venom Anim Toxins 4:5–21
Varanda EA, Monti R, Tavares DC (1999) Inhibitory effect of propolis and bee venom on the mutagenicity of some direct- and indirect-acting mutagens. Teratog Carcinog Mutagen 19:403–413. doi:10.1002/(SICI)1520-6866(1999)19:6<403:AID-TCM4>3.0.CO;2-2
Wang X, Feng Y, Wang N, Cheung F, Tan HY, Zhong S, Li C, Kobayashi S (2014) Chinese medicines induce cell death: the molecular and cellular mechanisms for cancer therapy. Biomed Res Int 2014:530342. doi:10.1155/2014/530342
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This work was supported by the Ministry of Science, Education and Sport of the Republic of Croatia (Grant Nos. 022-0222148-2125 and 098-0982913-2748).
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Gajski, G., Čimbora-Zovko, T., Rak, S. et al. Antitumour action on human glioblastoma A1235 cells through cooperation of bee venom and cisplatin. Cytotechnology 68, 1197–1205 (2016). https://doi.org/10.1007/s10616-015-9879-4
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DOI: https://doi.org/10.1007/s10616-015-9879-4