Evaluation of tumor-suppressive properties and apoptotic functions of Mad Honey and Vincristine applications in a rat model of breast cancer

Abstract

In this study, the suppressive effects of vincristine and Turkish mad honey alone and in co-applications were biochemically, hematologically, and histopathologically investigated in a mammary tumor model induced with 7,12-dimethylbenz[a]anthracene (DMBA) in rats. A total of 72 rats, 43-49 days old, were divided into 6 groups of 12 rats each. The control group (CG) consisted of healthy rats. The vehicle group (VG) received only vehicle substance and the cancer control group (CCG) was given only DMBA. DMBA and the honey group (HG) given group. DMBA and the vincristine (VinG) given group, and DMBA, the vincristine-honey group (VHG) received both Turkish mad honey and vincristine. Turkish mad honey and/or vincristine was given in the last 4 weeks of the 13-week trial period. White blood cell and lymphocyte counts differed significantly in the CCG and VG groups. Alanine transaminase and total protein levels were higher in the CCG and VinG groups. Aspartate transaminase was higher in the CCG, HG and VG groups. Caspase-3 and Bax protein levels were in the HG and VHG groups significantly higher than CCG. In caspase-8 protein level VHG significantly higher than other groups. Caspase -9 protein level was in CG and VG groups significantly lower than other groups. Bcl-xL increased more in the CCG group. Anaplasia was reduced in the HG and VinG groups, although apoptosis and other cellular damages increased. It was concluded that mad honey and vincristine could be considered together as effective therapeutic agents in this model of DMBA-induced breast cancer.

Keywords

• Apoptosis
• Breast Cancer
• Rat
• Turkish Mad Honey

References

1. Alzahrani HA, Boukraa L, Bellik Y, et al (2012): Evaluation of the Antioxidant Activity of Three Varieties of Honey from Different Botanical and Geographical Origins. Glob J Health Sci, 4, 192-196.
2. Bargou RC, Wagener C, Bommert K, et al (1996): Overexpression of the death-promoting gene bax-alpha which is downregulated in breast cancer restores sensitivity to different apoptotic stimuli and reduces tumor growth in SCID mice. J Clin Invest, 97, 2651-2659.
3. Beaudu-Lange C, Larrat S, Lange E, et al (2021): Prevalence of Reproductive Disorders including Mammary Tumors and Associated Mortality in Female Dogs. Vet Sci, 8, 184.
4. Blanc C, Deveraux QL, Krajewski S, et al (2000): Caspase-3 is essential for procaspase-9 processing and cisplatin-induced apoptosis of MCF-7 breast cancer cells. Cancer Res, 60, 4386-4390.
5. Blazquez S, Sirvent JJ, Olona M, et al (2006): Caspase-3 and caspase-6 in ductal breast carcinoma: a descriptive study. Histol Histopathol, 21, 1321-1329.
6. Buratti S, Benedetti S, Cosio MS (2007): Evaluation of the antioxidant power of honey, propolis and royal jelly by amperometric flow injection analysis. Talanta. 71, 1387-1392.
7. Cavalcanti C, Costa-Lotufo LV, Moraes MO, et al (2006): Genotoxicity evaluation of kaurenoic acid, a bioactive diterpenoid present in Copaiba oil. Food Chem Toxicol, 44, 338-392.
8. Chen J, Li S, Shen Q, et al (2011): Enhanced cellular uptake of folic acid-conjugated PLGA-PEG nanoparticles loaded with vincristine sulfate in human breast cancer. Drug Dev Ind Pharm, 37, 1339–1346.

9. Chipuk JE, Moldoveanu T, Llambi F, et al (2010): The BCL-2 family reunion. Mol Cell, 37, 299-310.
10. Christou M, Savas U, Schroeder S, et al (1995) Cytochromes CYP1a1 and CYP1B1 in the rat mammary gland: cell specific expression and regulation by polycyclic aromatic hydrocarbons and hormones. Mol Cell Endocrinol, 115, 41-50.
11. Danial NN, Korsmeyer SJ (2004): Cell death: critical control points. Cell, 116, 205-219.
12. Farombi EO, Shrotriya S, Surh YJ (2009): Kolaviron inhibits dimethyl nitrosamine-induced liver injury by suppressing COX-2 and iNOS expression via NF-ÍB and AP-1. Life Sci, 84,149-155.
13. Goldschmidt M, Pena L, Rasotto R, et al (2011): Classification and grading of canine mammary tumors. Vet Pathol, 48, 117-131.
14. Guillen KP, Fujita M, Butterfield AJ, et al (2022): A human breast cancer-derived xenograft and organoid platform for drug discovery and precision oncology. Nat Cancer, 3, 232–250.
15. Gulbahce ME, Baltaci S, Oztekin M, et al (2021): The Effect of Zinc and Melatonin Administration on Lipid Peroxidation, IL-6 Levels, and Element Metabolism in DMBA-Induced Breast Cancer in Rats. Biol Trace Elem Res, 199.
16. Holen I, Speirs V, Morrissey B, et al (2017): In vivo models in breast cancer research: progress, challenges and future directions. Dis Models Mech, 10, 359-371.
17. Jansen SA, Kleerekooper I, Hofman ZL, et al (2012): Grayanotoxin poisoning: ‘mad honey disease’ and beyond. Cardiovasc Toxicol, 12, 208-215.
18. Kabel AM (2017): Tumor markers of breast cancer: New prospectives. J Oncol Sci, 3, 5-11.
19. Kayaselcuk F, Nursal TZ, Polat A, et al (2004): Expression of survivin, bcl-2, P53 and bax in breast carcinoma and ductal intraepithelial neoplasia (DIN 1a). J Exp Clin Cancer Res, 23, 105-112.
20. Kennedy SM, O’Driscoll L, Purcell R, et al (2003) Prognostic importance of survivin in breast cancer. Br J Cancer, 88, 1077-1083.
21. LaCasse EC, Baird S, Korneluk RG, et al (1998): The inhibitors of apoptosis (IAPs) and their emerging role in cancer. Oncogene, 17, 3247-3259.
22. Luna LG (1968): Manual of Histologic Staining Methods; Armed Forces Institute of Pathology (U.S.); Editor, Lee G. Luna; Edition, 3; Publisher, McGraw-Hill, 1968.
23. Mohamed AM, Abdalla MS, Rizk MZ, et al (2014): Alleviation of Dimethylnitrosamine-induced liver injury and fibrosis by supplementation of Anabasis articulata extract in rats. Indian J Clin Biochem, 29, 418-429.
24. Moulder S, Hortobagyi GN (2008): Advances in the treatment of breast cancer. Clin Pharmacol Ther, 83, 26-36.
25. Nandakumar N, Balasubramanian MP (2011): Hesperidin protects renal and hepatic tissues against free radical-mediated oxidative stress during DMBA-induced experimental breast cancer. J Environ Pathol Toxicol Oncol, 30, 283-300.
26. Narimane S, Demircan E, Akkal S, et al (2017): Correlation between antioxidant activity and phenolic acids profile and content of Algerian propolis: Influence of solvent. Pak J Pharm Sci, 30, 1417-1423.
27. Oh JW, Yang WI, Lee MJ, et al (2009): The prognostic significance of survivin expression in breast cancer. J Breast Cancer, 12, 285-294.
28. Ohkawa H, Ohishi N, Yagi K (1979): Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem, 195, 351-358.
29. Okada E, Murai Y, Matsui K, et al (2001): Survivin expression in tumor cell nuclei is predictive of a favorable prognosis in gastric cancer patients. Cancer Lett, 163, 109-116.
30. Ozdemir H, Oto G, Ekin S, et al (2017): The protective effects of Lepidium sativum l. In 7,12-dimethylbenz(a)anthracene applied rats. Fresenius Environ Bull, 26, 2446-2453.
31. Pakkiri P, Lakhani SR, Smart CE (2009): Current and future approach to the pathologist’s assessment for targeted therapy in breast cancer. Pathology, 4189-4199.
32. Pu X, Storr SJ, Zhang Y, et al (2017): Caspase-3 and caspase-8 expression in breast cancer: caspase-3 is associated with survival. Apoptosis, 22, 357-368.
33. Russo J, Russo IH (2000): Atlas and histologic classification of tumors of the rat mammary gland. J Mammary Gland Biol Neoplasia, 5, 187-200.
34. Ryan BM, Konecny GE, Kahlert S, et al (2006): Survivin expression in breast cancer predicts clinical outcome and is associated with HER2, VEGF, urokinase plasminogen activator and PAI-1. Ann Oncol, 17, 597-604.
35. Sahin A, Türkmen S, Güzel N, et al (2018): A Comparison of the Effects of GrayanotoxinContaining Honey (Mad Honey), Normal Honey, and Propolis on Fracture Healing. Med Princ Pract, 27, 99-106.
36. Sharifi S, Barar J, Hejazi MS, et al (2014): Roles of the Bcl-2/Bax ratio, caspase-8 and 9 in resistance of breast cancer cells to paclitaxel. Asian Pac J Cancer Prev, 15, 8617-8622.
37. Sheikh A, Hussain SA, Ghori Q, et al (2015): The spectrum of genetic mutations in breast cancer. Asian Pac J Cancer Prev, 16, 2177-2185.
38. Shi Y (2002): Mechanisms of caspase activation and inhibition during apoptosis. Mol Cell, 9, 459-470.
39. Shokouh TZ, Ezatollah A, Barand P (2015): Interrelationships Between Ki67, HER2/neu, p53, ER, and PR Status and Their Associations With Tumor Grade and Lymph Node Involvement in Breast Carcinoma Subtypes: Retrospective-Observational Analytical Study. Medicine (Baltimore), 94, 1359.
40. Sun Y, Oberley LW, Li Y (1988): A simple method for clinical assay of superoxide dismutase. Clin Chem, 34, 497-500.
41. Tanaka K, Iwamoto S, Gon G, et al (2000): Expression of survivin and its relationship to loss of apoptosis in breast carcinomas. Clin Cancer Res, 6, 127-134.
42. Testa U, Castelli G, Pelosi E (2020): Breast Cancer: A Molecularly Heterogenous Disease Needing Subtype-Specific Treatments. Med Sci (Basel), 8, 18.
43. Thomas E, Holmes FA, Smith TL, et al (2004): The use of alternate, non-cross-resistant adjuvant chemotherapy on the basis of pathologic response to a neoadjuvant doxorubicin-based regimen in women with operable breast cancer: long-term results from a prospective randomized trial. J Clin Oncol, 22, 2294-2302.
44. Wolf EJ, Harrington KM, Clark SL, et al (2013): Sample Size Requirements for Structural Equation Models: An Evaluation of Power, Bias, and Solution Propriety. Educational and Psychological Measurement, 6, 913–934.