Enhancement of Doxorubicin Cytotoxicity by Verapamil in Human Breast Cancer Cells

Main Article Content

Sameer E. Al-harthy
Mashael S. Al-Motairi
Huda M. Al-Kreathy
Fatemah O. Kamel
Mohamed M. Sayed-Ahmed
Etimad Abbas Huwait
Hamdan S. Al-Malkey
Abdel-Moneim M. Osman

Abstract

Background: Worldwide, breast cancer is a main cause of morbidity and mortality in females. Doxorubicin (DOX) is an anthracycline anticancer drug and most commonly employed in polychemotherapy protocols in the treatment of solid and hematological tumors. Unfortunately, its optimal clinical benefit is limited secondary to the rapid development of DOX resistance and therapeutic failure.

Aim: Therefore, the current study has been initiated to investigate the possible mechanisms whereby the calcium channel blocker Verapamil (VER) could decrease DOX resistance and enhance the cytotoxic activity of DOX against the growth of human breast cancer cells.

Methodology: To achieve the ultimate goal of this study, we have examined DOX-induced cytotoxicity, apoptosis, alteration in the function of multidrug resistance proteins and cell cycle phase distribution against MCF-7 cell line in presence and absence of Verapamil. 

Results: Addition of VER enhanced the cytotoxic effect of DOX against the growth of MCF-7 cells which manifested as a significant decrease in the IC50 from 36 µg/ml for DOX alone to 13 µg/ml for DOX plus VER.  Moreover, combined treatment with VER and DOX significantly increased percentage of early apoptosis and cells arrested in G0/G1 phase when compared to DOX alone. In addition, VER significantly increased DOX cellular uptake through inhibition of the function of multidrug resistant proteins.  

Conclusion: VER treatment enhanced the cytotoxic activity of DOX against the growth of MCF-7 cells secondary to increase its cellular accumulation.  The observed increase in DOX uptake by VER was parallel to increased accumulation of Rho-123 dye which my point to the contribution of inhibition of multidrug resistant proteins by VER in the enhancement of DOX cytotoxicity.

Keywords:
Doxorubicin, verapamil, cytotoxicity, potentiation, breast cancer cells.

Article Details

How to Cite
Al-harthy, S., Al-Motairi, M., Al-Kreathy, H., Kamel, F., Sayed-Ahmed, M., Huwait, E., Al-Malkey, H., & Osman, A.-M. (2019). Enhancement of Doxorubicin Cytotoxicity by Verapamil in Human Breast Cancer Cells. Journal of Pharmaceutical Research International, 27(6), 1-10. https://doi.org/10.9734/jpri/2019/v27i630188
Section
Original Research Article

References

Al-Madouj AN, Al-Zahrani AS, Ao SF. Cancer incidence among nationals of the GCC states, 1998–2009. Riyadh, Saudi Arabia: King Faisal Specialist Hospital and Research Center; 2013.

Liu J, Mao W, Ding B, Liang CS. ERKs/p 53 signal transduction pathway is involved in doxorubicin-induced apoptosis in H9c2 cells and cardiomyocytes. American Journal of Physiology-Heart and Circulatory Physiology, 2008;295(5):H1956 -H1965.

Geisberg CA, Sawyer DB. Mechanisms of anthracycline cardiotoxicity and strategies to decrease cardiac damage. Current Hypertension Reports. 2010;12(6):404-410.
DOI:10.1007/s11906-010-0146

Brunton L, Knollmann B, Hilal-Dandan R. Goodman & Gilman's the pharmacological basis of therapeutics. 11th ed. McGraw-Hill's; 2005.

Wang X, Yang L, Chen Z, Shin DM. Application of nanotechnology in cancer therapy and imaging. CA: A cancer journal for clinicians. 2008;58(2):97-110.

Timcheva CV, Todorov DK. Does verapamil help overcome multidrug resistance in tumor cell lines and cancer patients? Journal of Chemotherapy. 1996;8(4):295-299.

Mason RP. Effects of calcium channel blockers on cellular apoptosis: implications for carcinogenic potential. Cancer, 1999;85(10):2093-2102.

Osman AMM, Mohamad MA, Abdel-Wahab AHA., Sayed-Ahmad MM. Modulation by verapamil of doxorubicin induced expression of multidrug resistance gene (mdr-1/P-glycoprotein) in murine tumour cells. J Egypt Natl Cancer Inst. 1995;12:221-227.

Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, Boyd MR. New colorimetric cytotoxicity assay for anticancer-drug screening. JNCI: Journal of the National Cancer Institute, 1990;82(13):1107-1112.

Pozarowski P, Darzynkiewicz Z. Analysis of cell cycle by flow cytometer. Methods Mol boil. 2004;281:301-311.

Bachur ANR, Cradock JC. Daunomycin metabolism in rat tissue slices. Journal of Pharmacology and Experimental Therapeutics. 1970;175(2):331-337.

Sayed-Ahmed MM. Multidrug resistance to cancer chemotherapy: Genes involved and blockers. Saudi Pharmaceutical Journal 2007;15(3-4):161-175.

Middleman E, Luce J, Frei Iii E. Clinical trials with adriamycin. Cancer, 1971;28(4): 844-850.

Von Hoff DD, Layard MW, Basa P, Davis HL, Von Hoff AL, Rozencweig M, Muggia FM. Risk factors for doxorubicin-lnduced congestive heart failure. Annals of Internal Medicine. 1979;91(5):710-717.

Singal PK, Deally CMR, Weinberg LE. Subcellular effects of adriamycin in the heart: a concise review. Journal of Molecular and Cellular Cardiology. 1987;19(8):817-828.

Fernandes G, Barone A, Dziak R. Effects of verapamil on bone cancer cells. Journal of Cell-Biology-&-Cell-Metabolism. 2016;3: 13.

Loe DW, Deeley RG, Cole SPC. Verapamil stimulates glutathione transport by the 190-kDa multidrug resistance protein 1 (MRP1). Journal of Pharmacology and Experimental Therapeutics, 2000;293(2): 530-538.

Perrotton T, Trompier D, Chang XB, Di Pietro A, Baubichon-Cortay H. (R)-and (S)-verapamil differentially modulate the multidrug-resistant protein MRP1. Journal of Biological Chemistry, 2007;282(43): 31542-31548.‏

Jensen RL, Lee YS, Guijrati M, Origitano TC, Wurster RD, Reichman OH. Inhibition of in vitro meningioma proliferation after growth factor stimulation by calcium channel antagonists: Part II-Additional growth factors, growth factor receptor immunohistochemistry, and intracellular calcium measurements. Neurosurgery, 1995;37(5):937-947.

Zheng W, Li M, Lin Y, Zhan X. Encapsulation of verapamil and doxorubicin by MPEG-PLA to reverse drug resistance in ovarian cancer. Biomedicine & Pharmacotherapy, 2018;108:565-573.

Huber KR, Schmidt WF, Thompson EA, Forsthoefel AM, Neuberg RW, Ettinger RS. Effect of verapamil on cell cycle transit and c-myc gene expression in normal and malignant murine cells. British Journal of Cancer. 1989;59(5):714-718.

Cao QZ, Niu G, Tan HR. In vitro growth inhibition of human colonic tumor cells by Verapamil. World J Gastroenterol, 2005;11(15):2255-2259. DOI:10.3748/wjg.v11.i15.2255.

Walworth NC. Cell-cycle checkpoint kinases: checking in on the cell cycle. Current Opinion in Cell Biology. 2000;12(6):697-704.

Zhou BBS, Elledge SJ. The DNA damage response: Putting checkpoints in perspective. Nature. 2000;408(6811):433.

Balakrishnan B, Dhandapani M, Pitchaikutti S, Sivamani G. Improving the efficacy of cyclophosphamide by using verapamil as a P-glycoprotein inhibitor in breast carcinoma. Journal of Drug Delivery and Therapeutics. 2018;8(5):349-353.

Giang I, Boland EL, Poon GM. Prodrug applications for targeted cancer therapy. Aaps j. 2014;16(5):899-913.
DOI:10.1208/s12248-014-9638

Goldie JH. Drug resistance in cancer: A perspective. Cancer and Metastasis Reviews. 2001;20(1-2):63-68.

Driscoll L, Clynes M. Biomarkers and multiple drug resistance in breast cancer. Current Cancer Drug Targets. 2006;6(5): 365-384.

Kibria G, Hatakeyama H, Harashima H. Cancer multidrug resistance: Mechanisms involved and strategies for circumvention using a drug delivery system. Archives of pharmacal research. 2014;37(1):4-15.

Minotti G, Menna P, Salvatorelli E, Cairo G, Gianni L. Anthracyclines: Molecular advances and pharmacologic developments in antitumor activity and cardiotoxicity. Pharmacological Reviews. 2004;56(2):185-229.

Sarkadi B, Homolya L, Szakács G, Váradi A. Human multidrug resistance ABCB and ABCG transporters: Participation in a chemoimmunity defense system. Physiological Reviews. 2006;86(4):1179-1236.

Solomon R, Gabizon AA. Clinical pharmacology of liposomal anthracyclines: Focus on pegylated liposomal doxorubicin. Clinical Lymphoma and Myeloma. 2008;8(1):21-32.