Naringenin Enhances the Anti-Cancer Effect of Cyclophosphamide against MDA-MB-231 Breast Cancer Cells Via Targeting the STAT3 Signaling Pathway

Document Type : Research article

Authors

1 Department of Biochemistry, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

2 Proteomics Research Center, School of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

3 Department of Pharmaceutical Chemistry, School of Pharmacy, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Abstract

Naringenin is a natural compound with potential anti-cancer effects against several cancer types.  Also, its precise molecular mechanisms regarding tumor growth suppression has not been completely elucidated. In the current study the apoptosis-inducing and anti-proliferative effects of Naringenin together with cyclophosphamide were studied in breast cancer cells and the participation of JAK2/STAT3 pathway was investigated. In this regard, MDA-MB-231 breast cancer cells were cultured and hence, treated with different concentrations of Naringenin. Apoptosis was measured via flowcytometric analysis of annexin V binding and cell viability was assessed via MTT assay. Protein and gene expression were investigated via Western blotting and real-time PCR, respectively. The function of caspase enzymes were also assessed. The results exhibited that Naringenin triggered apoptosis and markedly decreased cell viability. Furthermore its coadministration with cyclophosphamide improved its anti-tumor properties. Moreover, Naringenin up-regulated the expression of BAX while decreased the expression of Bcl-2. Caspases 3 and 9 were activated by Naringenin, an influence, which was augmented via cyclophosphamide. Docking studies revealed an interaction between Naringenin and STAT3 that was confirmed via attenuation of STAT3 phosphorylation subsequent to treating the cells with Naringenin. Furthermore, Naringenin exhibited the capacity to suppress the function of IL-6 in modulating apoptosis-associated genes expression. Overall, these results indicated that a Naringenin- cyclophosphamide combination impairs proliferation signaling and induces apoptosis to a greater extent than either compound alone and can serve as a potent chemotherapeutic regimen for breast cancer treatment.

Graphical Abstract

Naringenin Enhances the Anti-Cancer Effect of Cyclophosphamide against MDA-MB-231 Breast Cancer Cells Via Targeting the STAT3 Signaling Pathway

Keywords

References

(1)          Wang R, Wang J, Dong T, Shen J, Gao X, and Zhou J. Naringenin has a chemoprotective effect in MDA-MB-231 breast cancer cells via inhibition of caspase-3 and -9 activities. Oncology letters. (2019) 17: 1217-22.
(2)          Koh SY, Moon JY, Unno T, and Cho SK. Baicalein Suppresses Stem Cell-Like Characteristics in Radio- and Chemoresistant MDA-MB-231 Human Breast Cancer Cells through Up-Regulation of IFIT2. Nutrients. (2019) 11: 624.
(3)          Cella D and Fallowfield LJ. Recognition and management of treatment-related side effects for breast cancer patients receiving adjuvant endocrine therapy. Breast Cancer Res Treat. (2008) 107: 167-80.
(4)          Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ, and He J. Cancer statistics in China, 2015. CA Cancer J Clin. (2016) 66: 115-32.
(5)          Taylor CW and Kirby AM. Cardiac Side-effects From Breast Cancer Radiotherapy. Clin Oncol (R Coll Radiol). (2015) 27: 621-9.
(6)          Park SE, Shin WT, Park C, Hong SH, Kim GY, Kim SO, Ryu CH, Hong SH, and Choi YH. Induction of apoptosis in MDA-MB-231 human breast carcinoma cells with an ethanol extract of Cyperus rotundus L. by activating caspases. Oncol Rep. (2014) 32: 2461-70.
(7)          Ravishankar D, Rajora AK, Greco F, and Osborn HM. Flavonoids as prospective compounds for anti-cancer therapy. Int J Biochem Cell Biol. (2013) 45: 2821-31.
(8)          Song HM, Park GH, Eo HJ, Lee JW, Kim MK, Lee JR, Lee MH, Koo JS, and Jeong JB. Anti-Proliferative Effect of Naringenin through p38-Dependent Downregulation of Cyclin D1 in Human Colorectal Cancer Cells. Biomolecules & therapeutics. (2015) 23: 339-44.
(9)          Amaro M, Rocha J, Vila-Real H, Figueira M, Mota-Filipe H, Sepodes B, and Ribeiro M. Anti-inflammatory activity of naringin and the biosynthesised naringenin by naringinase immobilized in microstructured materials in a model of DSS-induced colitis in mice. Food Research International. (2009) 42:
(10)        Renugadevi J and Prabu SM. Naringenin protects against cadmium-induced oxidative renal dysfunction in rats. Toxicology. (2008) 256: 128-34.
(11)        Ahamad MS, Siddiqui S, Jafri A, Ahmad S, Afzal M, and Arshad M. Induction of apoptosis and antiproliferative activity of naringenin in human epidermoid carcinoma cell through ROS generation and cell cycle arrest. PLoS One. (2014) 9: e110003.
(12)        Wang B-d, Yang Z-Y, Wang Q, Cai T-k, and Crewdson P. Synthesis, characterization, cytotoxic activities, and DNA-binding properties of the La(III) complex with Naringenin Schiff-base. Bioorganic & Medicinal Chemistry. (2006) 14: 1880-8.
(13)        Kanno S-I, Tomizawa A, Hiura T, Osanai Y, Shouji A, Ujibe M, Ohtake T, Kimura K, and Ishikawa M. Inhibitory Effects of Naringenin on Tumor Growth in Human Cancer Cell Lines and Sarcoma S-180-Implanted Mice. Biological & pharmaceutical bulletin. (2005) 28: 527-30.
(14)        Kanno S-i, Tomizawa A, Ohtake T, Koiwai K, Ujibe M, and Ishikawa M. Naringenin-induced apoptosis via activation of NF-κB and necrosis involving the loss of ATP in human promyeloleukemia HL-60 cells. Toxicology Letters. (2006) 166: 131-9.
(15)        Emadi A, Jones RJ, and Brodsky RA. Cyclophosphamide and cancer: golden anniversary. Nature reviews Clinical oncology. (2009) 6: 638.
(16)        Stoll B. Evaluation of cyclophosphamide dosage schedules in breast cancer. British journal of cancer. (1970) 24: 475.
(17)        Ahmed AR and Hombal SM. Cyclophosphamide (Cytoxan): a review on relevant pharmacology and clinical uses. Journal of the American Academy of Dermatology. (1984) 11: 1115-26.
(18)        Elazzazy S, Mohamed AE, and Gulied A. Cyclophosphamide-induced symptomatic hyponatremia, a rare but severe side effect: a case report. OncoTargets and therapy. (2014) 7: 1641-5.
(19)        Park JH, Im S-A, Byun JM, Kim KH, Kim J-S, Choi IS, Kim H-J, Lee K-H, Kim T-Y, Han S-W, Oh DY, and Kim T-Y. Cyclophosphamide, Methotrexate, and 5-Fluorouracil as Palliative Treatment for Heavily Pretreated Patients with Metastatic Breast Cancer: A Multicenter Retrospective Analysis. Journal of breast cancer. (2017) 20: 347-55.
(20)        Emadi A, Jones RJ, and Brodsky RA. Cyclophosphamide and cancer: golden anniversary. Nat Rev Clin Oncol. (2009) 6: 638-47.
(21)        Masjedi A, Hashemi V, Hojjat-Farsangi M, Ghalamfarsa G, Azizi G, Yousefi M, and Jadidi-Niaragh F. The significant role of interleukin-6 and its signaling pathway in the immunopathogenesis and treatment of breast cancer. Biomed Pharmacother. (2018) 108: 1415-24.
(22)        Judd LM, Menheniott TR, Ling H, Jackson CB, Howlett M, Kalantzis A, Priebe W, and Giraud AS. Inhibition of the JAK2/STAT3 Pathway Reduces Gastric Cancer Growth In Vitro and In Vivo. PLOS ONE. (2014) 9: e95993.
(23)        Becker S, Groner B, and Muller CW. Three-dimensional structure of the Stat3beta homodimer bound to DNA. Nature. (1998) 394: 145-51.
(24)        Quispe-Soto ET and Calaf GM. Effect of curcumin and paclitaxel on breast carcinogenesis. Int J Oncol. (2016) 49: 2569-77.
(25)        Huang L, Li A, Liao G, Yang F, Yang J, Chen X, and Jiang X. Curcumol triggers apoptosis of p53 mutant triple-negative human breast cancer MDA-MB 231 cells via activation of p73 and PUMA. Oncol Lett. (2017) 14: 1080-8.
(26)        Yuan J, Zhang F, and Niu R. Multiple regulation pathways and pivotal biological functions of STAT3 in cancer. Scientific Reports. (2015) 5: 17663.
(27)        Braun DA, Fribourg M, and Sealfon SC. Cytokine response is determined by duration of receptor and signal transducers and activators of transcription 3 (STAT3) activation. J Biol Chem. (2013) 288: 2986-93.
(28)        Nicholson RI and Johnston SR. Endocrine therapy--current benefits and limitations. Breast Cancer Res Treat. (2005) 93 Suppl 1: S3-10.
(29)        Kim S and Park TI. Naringenin: a partial agonist on estrogen receptor in T47D-KBluc breast cancer cells. Int J Clin Exp Med. (2013) 6: 890-9.
(30)        Lee J, Kim D-H, and Kim JH. Combined administration of naringenin and hesperetin with optimal ratio maximizes the anti-cancer effect in human pancreatic cancer via down regulation of FAK and p38 signaling pathway. Phytomedicine. (2019) 58: 152762.
(31)        Hatkevich T, Ramos J, Santos-Sanchez I, and Patel YM. A naringenin-tamoxifen combination impairs cell proliferation and survival of MCF-7 breast cancer cells. Exp Cell Res. (2014) 327: 331-9.
(32)        Baig S, Seevasant I, Mohamad J, Mukheem A, Huri HZ, and Kamarul T. Potential of apoptotic pathway-targeted cancer therapeutic research: Where do we stand? Cell death & disease. (2016) 7: e2058-e.
(33)        Arul D and Subramanian P. Naringenin (citrus flavonone) induces growth inhibition, cell cycle arrest and apoptosis in human hepatocellular carcinoma cells. Pathol Oncol Res. (2013) 19: 763-70.
(34)        Hernández-Aquino E and Muriel P. Beneficial effects of naringenin in liver diseases: Molecular mechanisms. World journal of gastroenterology. (2018) 24: 1679-707.
(35)        Lim W, Park S, Bazer FW, and Song G. Naringenin-Induced Apoptotic Cell Death in Prostate Cancer Cells Is Mediated via the PI3K/AKT and MAPK Signaling Pathways. J Cell Biochem. (2017) 118: 1118-31.
Iranian Journal of Pharmaceutical Research
Volume 19, Issue 3
Summer 2020
Pages 122-133

Files

Share

How to cite

Statistics