Investigation of The Synergistic Effects of Trastuzumab And Gambogic Acid in Her-2 Positive Breast Cancer Cell Line

Abstract

HER2 positive breast cancer is one of the biggest health problems in the world, causing millions of deaths every year. Drug combination modeling studies are extensively evaluated in treating many diseases. Pharmacological studies over the last half-century have shown that gambogic acid has potent anti-tumor activity against many types of cancer, including breast cancer. In this study, we examined the synergistic anticancer effect of gambogic acid and trastuzumab in HER2 positive breast cancer cell line (MDA-MB-453). In-vitro synergistic and antiproliferative effects of trastuzumab plus gambogic acid studies were determined with XTT method and the combination index (CI) values of the trastuzumab and gambogic acid combination were calculated by CompuSyn software. To determine molecular mechanisms of the trastuzumab and gambogic acid combination, we analyzed HER2, caspase-9 and Bax gene and protein expression levels quantitative reverse transcription-PCR (qRT-PCR) and ELISA techniques. The combination of 50 µg/ml trastuzumab and 5 µM gambogic acid showed the best synergistic effect at 24 h incubation in MDA-MB-453 cells according to the in-vitro cell proliferation, RT-qPCR and ELISA test. Gambogic acid effects on HER2 positive breast cancer cell line shows its potential as natural compound to inhibit breast cancer cell proliferation in combination with trastuzumab.

Keywords

• Trastuzumab,gambogic acid,breast cancer

References

1. 1. F. Bray, J. Ferlay, I. Soerjomataram, R.L. Siegel, L.A. Torre, A. Jemal, Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin., 68 (2018) 394-424.
2. 2. H.M. Asif, S. Sultana, S. Ahmed, N. Akhtar, M. Tariq, HER-2 Positive Breast Cancer - a Mini-Review. Asian Pac J Cancer Prev., 17(4) (2016) 1609-1615.
3. 3. S. Pernas, S.M. Tolaney, HER2-positive breast cancer: new therapeutic frontiers and overcoming resistance. Ther Adv Med Oncol., (2019) 11.
4. 4. W. Dean-Colomb, F.J. Esteva, Her2-positive breast cancer: herceptin and beyond. Eur J Cancer., 44 (2008) 2806-2812.
5. 5. S. Shak, Overview of the trastuzumab (Herceptin) anti-HER2 monoclonal antibody clinical program in HER2-overexpressing metastatic breast cancer. Herceptin Multinational Investigator Study Group. Semin Oncol., 26 (1999) 71-77.
6. 6. Y. Okawa, K. Sugiyama, K. Aiba, A. Hirano, S. Uno, T. Hagino, K. Kawase, H. Shioya, K. Yoshida, N. Usui, M. Kobayashi, T. Kobayashi, Successful combination therapy with trastuzumab and Paclitaxel for adriamycin- and docetaxel-resistant inflammatory breast cancer. Breast Cancer., 11 (2004) 309-312.
7. 7. E.A. Perez, Carboplatin in combination therapy for metastatic breast cancer. Oncologist., 9 (2004) 518-527.
8. 8. M.S. van Ramshorst, E. van Werkhoven, I.A.M. Mandjes, M. Schot, J. Wesseling, M.T.F.D. Vrancken Peeters, J.M. Meerum Terwogt, M.E.M. Bos, H.M. Oosterkamp, S. Rodenhuis, S.C. Linn, G.S. Sonke, Trastuzumab in combination with weekly paclitaxel and carboplatin as neo-adjuvant treatment for HER2-positive breast cancer: The TRAIN-study. Eur J Cancer., 74 (2017) 47-54.

9. 9. N. Hayashi, N. Niikura, H. Yamauchi, S. Nakamura, N.T. Ueno, Adding hormonal therapy to chemotherapy and trastuzumab improves prognosis in patients with hormone receptor-positive and human epidermal growth factor receptor 2-positive primary breast cancer. Breast Cancer Res Treat., 137 (2013) 523-531.
10. 10. D. Kashyap, R. Mondal, H.S. Tuli, G. Kumar, A.K. Sharma, Molecular targets of gambogic acid in cancer: recent trends and advancements. Tumour Biol., 37 (2016) 12915-12925.
11. 11. K. Banik, C. Harsha, D. Bordoloi, B. Lalduhsaki Sailo, G. Sethi, H.C. Leong, F. Arfuso, S. Mishra, L. Wang, A.P. Kumar, A.B. Kunnumakkara, Therapeutic potential of gambogic acid, a caged xanthone, to target cancer. Cancer Lett., 416 (2018) 75-86.
12. 12. G.M. Huang, Y. Sun, X. Ge, X. Wan, C.B. Li, Gambogic acid induces apoptosis and inhibits colorectal tumor growth via mitochondrial pathways. World J Gastroenterol., 21 (2015) 6194-6205.
13. 13. X. Wang, W. Chen, Gambogic acid is a novel anti-cancer agent that inhibits cell proliferation, angiogenesis and metastasis. Anticancer Agents Med Chem., 12 (2012) 994-1000.
14. 14. Y.I. Chi, X.K. Zhan, H. Yu, G.R. Xie, Z.Z. Wang, W. Xiao, Y.G. Wang, F.X. Xiong, J.F. Hu, L. Yang, C.X. Cui, J.W. Wang, An open-labeled, randomized, multicenter phase IIa study of gambogic acid injection for advanced malignant tumors. Chin Med J (Engl)., 126 (2013) 1642-1646.
15. 15. M. Gümus, A. Ozgur, L. Tutar, A. Disli, I. Koca, Y.Tutar, Design, Synthesis, and Evaluation of Heat Shock Protein 90 Inhibitors in Human Breast Cancer and Its Metastasis. Curr Pharm Biotechnol., 17 (2016) 1231-1245.
16. 16. İ. Koca, A. Özgür, M. Er, M. Gümüş, K. Açikalin Coşkun, Y. Tutar, Design and synthesis of pyrimidinyl acyl thioureas as novel Hsp90 inhibitors in invasive ductal breast cancer and its bone metastasis. Eur J Med Chem., 122 (2016) 280-290.
17. 17. IV Bijnsdorp, E. Giovannetti, G.J. Peters, Analysis of drug interactions. Methods Mol Biol., 731 (2011) 421-434.
18. 18. M. Luque-Cabal, P. García-Teijido, Y. Fernández-Pérez, L. Sánchez-Lorenzo, I Palacio-Vázquez, Mechanisms Behind the Resistance to Trastuzumab in HER2-Amplified Breast Cancer and Strategies to Overcome It. Clin Med Insights Oncol., 10 (2016) 21-30.
19. 19. G. Li, J. Guo, B.Q. Shen, D.B. Yadav, M.X. Sliwkowski, L.M. Crocker, J.A. Lacap, G.D.L. Phillips, Mechanisms of Acquired Resistance to Trastuzumab Emtansine in Breast Cancer Cells. Mol Cancer Ther., 17 (2018) 1441-1453.
20. 20. S.J. Riedl, W. Li, Y. Chao, R. Schwarzenbacher, Y. Shi, Structure of the apoptotic protease-activating factor 1 bound to ADP. Nature., 434 (2005) 926-933.
21. 21. S.J. Riedl, G.S. Salvesen, The apoptosome: signalling platform of cell death, Nat. Rev. Mol. Cell Biol., 8 (2007) 405–413.
22. 22. H. Zou, W.J. Henzel, X. Liu, A. Lutschg, X. Wang, Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3, Cell 90., 5 (1997) 405–413.
23. 23. E.A. Perez, J. Cortés, A.M. Gonzalez-Angulo, J.M. Bartlett, HER2 testing: current status and future directions. Cancer Treat Rev., 40 (2014) 276–284.