Evaluation of the effect of Melittin on liver and renal biochemical markers and KI67 expression in mice with experimentally-induced breast cancer

Subject Areas : Journal of Comparative Pathobiology

F.  Dabbagh Moghaddam, ¹ (.Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran)
, S.  Hamedi ² (Department of Basic Sciences, Karaj Branch, Islamic Azad University, Karaj, Iran)
, P  Mortazavi ³ (Department of Pathology, Faculty of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.)
, M.  Nabiuni ⁴ (Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran)
N.  Hayati Roodbari, ⁵ (Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran)

Received: 2020-11-07 Accepted : 2020-11-07 Published : 2020-05-21

Keywords: Breast cancer, Melittin, KI67, 4T1 Cell line, mice,

Abstract :

Despite extensive progress in the production of anti-cancer drugs, attention to some natural products, such as Melittin from honey bee venom, is growing. Some studies have shown that this product has anti-cancer properties. To evaluate the effects of Melittin on function breast cancer indicators in inbred Balb/c mice as well as its effects on liver and renal enzymes, 4T1 cell lines were purchased from Pasteur Institute and were cultured and the induction of breast cancer was performed in the breast area of ​​mice. The 20-day treatment period was performed in 9 groups (including 6,3,1mg/kg body weight of Melittin, doxorubicin, cisplatin, PBS, DMSO, healthy control, negative control). At the end of the course, animal blood samples were taken then liver and renal biomarkers were measured after serum isolation. From the induced tumor tissue sample, after fixation in formalin, the immunohistochemical tissue slide was prepared and studied for KI67 expression. Results showed that renal (BUN) and liver biochemical markers (AST, ALT as well as ALP) had no significant changes in melittin treated group at the dose of 6 mg/kg compared to control group. The results of histopathology showed a decrease in tumor cell invasion in Melittin therapeutic groups, which was more evident at a dose of 6 mg/kg. In immunohistochemistry, KI67 expression did not show a significant change in all groups. The results of this study show the beneficial effect of melittin in preventing the growth of tumor cells without injuring the liver and kidneys compared to other common drugs.

References:

1. Nouri Daloui M. Tabarestani S. Molecular Genetics, Diagnosis and Treatment of Breast Cancer: A Review Article. Sabzevar University of Medical Sciences 2010;17(2):87-747


2. Yeo SW. Seo JC. Choi YH. Jang KJ. Induction of growth inhibition and apoptosis by bee venom in human breast carcinoma MCF-7 cells. Kor Acup Mox Soc 2003;20(3):45-62


3. Moon DO. Park SY. Lee KJ. Heo MS. Kim KC. Kim MO. Bee venom and melittin reduce proinflammatory mediators in lipopolysaccharide-stimulated BV2 microglia. Int Immunopharmacol 2007;7(8):1092-101


4. Kaiser T. Brennecke SP. Moses EK. Methylen etetrahydrofolate reductase polymorphism are not a risk factor for preeclampsia/eclampsia in Australian women. Gynecol Obstet 2000;50(2):100-102


5. Bazzo R. Tappin MJ. Pastore A. Harvey TS. Carver JA. Campebell ID. The structure of melittin. Eur J Biochem 1988;173:139e46


6. Malekzadeh R. Derakhshan MH. Malekzadeh Z. Gastric cancer in Iran: epidemiology and risk factors. Arch Iran Med 2009;12:576e83


7. D'Elia AV. Driul L. Giacomello R. Frequency of factor V, prothrombin and methylene tetrahydro folate reductase gene variants in preeclampsia. Gynecol Obstet Invest 2002;53(2):84-87


8. Bramwell VW. Somavarapu S. Outschoorn I. Alpar HO. Adjuvant action of melittin following intranasal immunisation with tetanus and diphtheria toxoids. J Drug Target 2003;11(8-10):525-30


9. Son DJ. Ha SJ. Song HS. Lim Y. Melittin inhibits vascular smooth muscle cell proliferation through induction of apoptosis via suppression of nuclear factor-κB and Akt activation and enhancement of apoptotic protein expression. Pharmacology and

Experimental Therapeutics 2006; 317(2):627-34

10. Jang MH. Shin MC. Lim S. Han SM. Park HJ. Shin I. Bee venom induces apoptosis and inhibits expression of cyclooxygenase-2 mRNA in human lung cancer cell line NCI-H1299. J Pharmacol Sci 2003;91(2):95-10411. Wang C. Chen T. Zhang N. Yang M. Li B. Lü X. Melittin, a major component of bee venom , sensitizes human hepatocellular carcinoma cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis by activating CaMKII-TAK1-JNK/p38 and inhibiting IκBα kinase-NFκB. J. Biolog Chem 2009;284(6):3804-13


11. Ip SW. Liao SS. Lin SY. Lin JP. Yang JS. Lin ML. The role of mitochondria in bee venom-induced apoptosis in human breast cancer MCF7 cells. In Vivo 2008;22(2):237-45


12. Huh J.E. Baek Y.H. Lee M.H. Choi D.Y. Park, D.S. Lee J.D. Bee venom inhibitstumor angiogenesis and metastasis by inhibiting tyrosine phosphorylation ofVEGFR-2 in LLC-tumor-bearing mice. Cancer Lett 2010;292:98–110


13. Shin S.H. Kim Y.H. Kim J.K. Park K.K. Anti-allergic effect of bee venom inan allergic rhinitis mouse model. Biol. Pharm. Bull 2014;37:1295–1300


14. Kwon Y.B. Lee J.D. Lee H.J. Han H.J. Mar W.C. Kang S.K. Beitz A.J. Lee J.H. Bee venom injection into an acupuncture point reduces arthritis associ-ated edema and nociceptive responses. Pain 2001;90:271–280


15. Ling C. Li B. Zhang C. Gu W. Li S. Huang X. (Anti-hepatocarcinoma effect of recombinant adenovirus carrying melittin gene). Chinese journal of hepatology 2004;12(12):741-4


16. Ciu S. Yu M. He Y, Xiao L. Wang F. Song C. Melittin prevents liver cancer cell metastasis through inhibition of the Rac1-dependent pathway. Hepatology 2008;47(6):1964-73


17. Uawonggul N. Thammasirira KS. Chaveerach A. Chuachan C. Daduang S. Plant extract activities against the fibroblast cell lysis by honey bee venom J Med Plants Res 2011;5(10):1978-86


18. Winder D. Günzburgb WH. Erfle V. Salmons B. Expression of antimicrobial peptides has an antitumour effect in human cells. Biochem Biophy ResCommun 1998;242(3):608-12


19. Putz T. Ramoner R. Gander H. Rahm A. Bartsch G. Thurnher M. Antitumor action and immune activation through cooperation of bee venom secretory phospholipase A2 and phosphatidylinositol-(3, 4)- bisphosphate. Cancer Immunol Immunother 2006;55(11):1374-83


20. Nosrati N. Bakovic M. Paliyath G. Molecular Mechanisms and Pathways as Targets for Cancer Prevention and Progression with Dietary Compounds. Molecular Sciences 2017;18:2050


21. Dabbagh Moghaddam F. Hamedi S. Dezfulian M. Anti-tumor effect of C-phycocyanin from Anabaena sp.ISC55 in inbred BALB/c mice injected with 4T1 breast cancer cell. Comparative clinical Pathology 2016;25(5):947-952


22. Moon DO. Park SY. Choi YH. Kim ND. Lee C. Kim GY. Melittin induces Bcl-2 and caspase-3-dependent apoptosis through downregulation of Akt phosphorylation in human leukemic U937 cells. Toxicon. 2008;51:112–120


23. Wang C. Chen T. Zhang N. Yang M. Li B. Lu X. Melittin a major component of bee venom, sensitizes human hepatocellular carcinoma cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis by activating CaMKII-TAK1-JNK/p38 and inhibiting IkappaBalpha kinase-NFkappaB. J. Biol. Chem. 2009;284:3804–3813


24. Wong RS. Apoptosis in cancer: from pathogenesis to treatment. J. Exp. Clin. Cancer Res. 2011;30;87


25. Jeong YJ. Choi Y. Shin JM. Cho HJ. Kang JH. Park KK. Melittin suppresses EGF-induced cell motility and invasion by inhibiting PI3K/Akt/mTOR signaling pathway in breast cancer cells. Food Chem. Toxicol 2014;68:218–225


26. Wang X. Xiong L. Yu G. Li D. Peng T. Luo D. Cathepsin S silencing induces apoptosis of human hepatocellular carcinoma cells. Am. J. Transl. Res 2015;7:100–110


27. Jo M. Park MH. Kollipara PS. An BJ. Song HS. Han SB. 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 2012;258:72–81


28. Park MH. Choi MS. Kwak DH. Oh KW. Yoon DY. Han SB. Anti-cancer effect of bee venom in prostate cancer cells through activation of caspase pathway via inactivation of NF-kappaB. Prostate 2011;71:801–812


29. Tu WC. Wu CC. Hsieh HL. Chen CY. Hsu SL. Honeybee venom induces calcium-dependent but caspase-independent apoptotic cell death in human melanoma A2058 cells. Toxicon. 2008;5:318–329


30. Choi KE. Hwang CJ. Gu SM. Park MH. Kim JH. Park JH. 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) 2014;6:2210–2228


31. Moon DO. Park SY. Heo MS. Kim KC. Park C. Ko WS. 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 2006;6:1796–1807


32. Orsolic N. Potentiation of bleomycin lethality in HeLa and V79 cells by bee venom. Arh. Hig. Rada Toksikol 2009;60:317–326.


33. Alonezi S. Tusiimire J. Wallace J. Dufton M. Parkinson J. Young L. Clements C. Park J. Jeon J. Metabolomic Profiling of the Effects of Melittin on Cisplatin Resistant and Cisplatin Sensitive Ovarian Cancer Cells Using Mass Spectrometry and Biolog Microarray Technology. Published online 2016 Oct 13. Metabolites 2016;6(4):35


34. Jang MH. Shin MC. Lim S. Han SM. Park HJ. Shin I. Bee venom induces apoptosis and inhibits expression of cyclooxygenase-2 mRNA in human lung cancer cell line NCI-H1299. J. Pharmacol. Sci 2003;91:95–104


35. Lee G. Bae H. Anti-Inflammatory Applications of Melittin, a Major Component of Bee Venom: Detailed Mechanism of Action and Adverse Effects. Molecules 2016;21:616


36. .Rady I. Siddiqui I. Rady M. Mukhtar H. Melittin, a major peptide component of bee venom, and its conjugates in cancer therapy. Cancer Lett 2017;28(402):16–23.