Abstract
Breast cancer is the most common malignancy in women worldwide. Strategies for cancer chemotherapy commonly require the use of combination therapy for better outcomes of results. The present work is aimed to evaluate the potential of naringenin and metformin concomitant addition with doxorubicin chemotherapy against experimental breast carcinoma. The antitumor potential of drugs under the study was evaluated against methylnitrosourea (MNU)-induced breast cancer in rats and 4T1 cells–induced orthotopic breast cancer mouse model. Parameters like tumor growth, body weight, survival rate, blood glucose, hematology, and histology were determined. There was a marked reduction in tumor weight and an observed decrease in tumor multiplicity by naringenin and metformin concomitant addition with doxorubicin against MNU-induced breast carcinoma. Likewise, naringenin and metformin with doxorubicin showed a significant reduction of tumor volume and tumor weight (p < 0.01) in 4T1-induced orthotopic mouse model as compared to the same dose of doxorubicin alone, suggesting combination treatment enhanced antitumor activity in vivo. Furthermore, histology of tumor biopsies presented the improved antitumor activity of doxorubicin via increasing tumor necrosis. Hematological parameters, body weight, and survival data presented remarkable safety of combination treatment without compromising efficacy using 50% lower dose of doxorubicin as compared to the large dose of doxorubicin alone. These results demonstrate that naringenin and metformin enhanced the antitumor effect of doxorubicin in animal models of breast carcinoma, and therefore can be useful as an adjunct treatment with doxorubicin to increase its effectiveness at the lower dose level for the treatment of cancer.
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References
Ajzashokouhi AH, Bostan HB, Jomezadeh V et al (2020) A review on the cardioprotective mechanisms of metformin against doxorubicin. Hum Exp Toxicol 39:237–248. https://doi.org/10.1177/0960327119888277
Aljofan M, Riethmacher D (2019) Anticancer activity of metformin: a systematic review of the literature. Futur Sci OA 5. https://doi.org/10.2144/fsoa-2019-0053
Alyahya R, Sudha T, Racz M et al (2015) Anti-metastasis efficacy and safety of non-anticoagulant heparin derivative versus low molecular weight heparin in surgical pancreatic cancer models. Int J Oncol 46:1225–1231. https://doi.org/10.3892/ijo.2014.2803
Bissery MC, Gueritte-Voegelein F (1991) Experimental antitumor activity of taxotere (RP 56976, NSC 628503), a Taxol Analogue. Cancer Res 51:4845–4852
Burade V, Bhowmick S, Maiti K et al (2017) Lipodox® (generic doxorubicin hydrochloride liposome injection): In vivo efficacy and bioequivalence versus Caelyx® (doxorubicin hydrochloride liposome injection) in human mammary carcinoma (MX-1) xenograft and syngeneic fibrosarcoma (WEHI 164) mouse mode. BMC Cancer 17:1–12. https://doi.org/10.1186/s12885-017-3377-3
El-Ashmawy NE, Khedr NF, El-Bahrawy HA, Abo Mansour HE (2017) Metformin augments doxorubicin cytotoxicity in mammary carcinoma through activation of adenosine monophosphate protein kinase pathway. Tumor Biol 39. https://doi.org/10.1177/1010428317692235
Faustino-Rocha AI, Ferreira R, Oliveira PA et al (2015) N-Methyl-N-nitrosourea as a mammary carcinogenic agent. Tumor Biol 36:9095–9117. https://doi.org/10.1007/s13277-015-3973-2
Fisusi FA, Akala EO (2019) Drug combinations in breast cancer therapy. Pharm Nanotechnol 7:3–23. https://doi.org/10.2174/2211738507666190122111224
Grossmann ME, Yang DQ, Guo Z et al (2015) Metformin treatment for the prevention and/or treatment of breast/mammary tumorigenesis. Curr Pharmacol Reports 1:312–323. https://doi.org/10.1007/s40495-015-0032-z
Hanušová V, Boušová I, Skálová L (2011) Possibilities to increase the effectiveness of doxorubicin in cancer cells killing. Drug Metab Rev 43:540–557. https://doi.org/10.3109/03602532.2011.609174
Harbeck N, Penault-Llorca F, Cortes J et al (2019) Breast cancer. Nat Rev Dis Prim 5:66. https://doi.org/10.1038/s41572-019-0111-2
Hirsch HA, Iliopoulos D, Tsichlis PN, Struhl K (2009) Metformin selectively targets cancer stem cells, and acts together with chemotherapy to block tumor growth and prolong remission. Cancer Res 69:7507–7511. https://doi.org/10.1158/0008-5472.CAN-09-2994
Kanno SI, Tomizawa A, Hiura T et al (2005) Inhibitory effects of naringenin on tumor growth in human cancer cell lines and sarcoma S-180-implanted mice. Biol Pharm Bull 28:527–530. https://doi.org/10.1248/bpb.28.527
Karia PD, Patil LA, Vakani MS et al (2018) Chemoprevention of breast cancer by Psidium guajava Linn. Asian J Pharm Pharmacol 5:58–68. https://doi.org/10.31024/ajpp.2019.5.1.8
Kasznicki J, Sliwinska A, Drzewoski J (2014) Metformin in cancer prevention and therapy. Ann Transl Med 2:1–11. https://doi.org/10.3978/j.issn.2305-5839.2014.06.01
Kruczynski A, Hill BT (2002) Classic in vivo cancer models: three examples of mouse models used in experimental therapeutics. Curr Protoc Pharmacol 1–16https://doi.org/10.1002/0471141755.ph0524s15
Lee KN, Torres MA, Troeschel AN et al (2020) Type 2 diabetes, breast cancer specific and overall mortality: associations by metformin use and modification by race, body mass, and estrogen receptor status. PLoS ONE 15:1–13. https://doi.org/10.1371/journal.pone.0232581
Li W, Wang QL, Liu X et al (2015) Combined use of vitamin D3 and metformin exhibits synergistic chemopreventive effects on colorectal neoplasia in rats and mice. Cancer Prev Res 8:139–148. https://doi.org/10.1158/1940-6207.CAPR-14-0128
Li Y, Wang M, Zhi P et al (2018) Metformin synergistically suppress tumor growth with doxorubicin and reverse drug resistance by inhibiting the expression and function of P-glycoprotein in MCF7/ADR cells and xenograft models. Oncotarget 9:2158–2174. https://doi.org/10.18632/oncotarget.23187
Liu X, Yang X, Chen F, Chen D (2017) Combined application of doxorubicin and naringin enhances the antitumor efficiency and attenuates the toxicity of doxorubicin in HeLa cervical cancer cells. Int J Clin Exp Pathol 10:7303–7311
Marinello PC, Panis C, Silva TNX et al (2019) Metformin prevention of doxorubicin resistance in MCF-7 and MDA-MB-231 involves oxidative stress generation and modulation of cell adaptation genes. Sci Rep 9:1–11. https://doi.org/10.1038/s41598-019-42357-w
Muthusamy T, Yadav LR, Ramalingam S (2020) Synergistic effect of 5-fluorouracil combined with naringin in MDA-MB-231 human breast cancer cells. Int Res J Oncol 3:13–27
Noori S, Tavirani MR, Deravi N et al (2020) Naringenin enhances the anti-cancer effect of cyclophosphamide against MDA-MB-231 breast cancer cells via targeting the STAT3 signaling pathway. Iran J Pharm Res 19:122–133. https://doi.org/10.22037/ijpr.2020.113103.14112
Parvathaneni M, Battu GR, Gray AI, Gummalla P (2014) Investigation of anticancer potential of hypophyllanthin and phyllanthin against breast cancer by in vitro and in vivo methods. Asian Pacific J Trop Dis 4https://doi.org/10.1016/S2222-1808(14)60417-5
Paschall AV, Liu K (2016) An Orthotopic mouse model of spontaneous breast cancer metastasis. J vis Exp 2016:1–7. https://doi.org/10.3791/54040
Pateliya B, Burade V, Goswami S (2021) Combining naringenin and metformin with doxorubicin enhances anticancer activity against triple-negative breast cancer in vitro and in vivo. Eur J Pharmacol 891:173725. https://doi.org/10.1016/j.ejphar.2020.173725
Salehi B, Fokou PVT, Sharifi-Rad M et al (2019) The therapeutic potential of naringenin: a review of clinical trials. Pharmaceuticals 12:1–18. https://doi.org/10.3390/ph12010011
Shabanah OA, Ahmed LA, Qunebet RA et al (2019) Losartan and/or naringenin ameliorates doxorubicin induced cardiac, hepatic and renal toxicities in rats. Int J Pharmacol 15:675–685. https://doi.org/10.3923/ijp.2019.675.685
Sheta A, Elsakkar M, Hamza M, Solaiman A (2016) Effect of metformin and sitagliptin on doxorubicin-induced cardiotoxicity in adult male albino rats. Hum Exp Toxicol 35:1227–1239. https://doi.org/10.1177/0960327115627685
Subburaman S, Ganesan K, Ramachandran M (2014) Protective role of naringenin against doxorubicin-induced cardiotoxicity in a rat model: histopathology and mRNA expression profile studies. J Environ Pathol Toxicol Oncol 33:363–376. https://doi.org/10.1615/JEnvironPatholToxicolOncol.2014010625
Thompson HJ, Adlakha H (1991) Dose-responsive induction of mammary gland carcinomas by the intraperitoneal injection of 1-methyl-l-nitrosourea. Cancer Res 51:3411–3415
Ugwueze C V., Ogamba OJ, Young EE, et al (2020) Metformin: a possible option in cancer chemotherapy. Anal Cell Pathol 2020. https://doi.org/10.1155/2020/7180923
Viollet B, Guigas B, Sanz Garcia N et al (2012) Cellular and molecular mechanisms of metformin: an overview. Clin Sci 122:253–270. https://doi.org/10.1042/CS20110386
Wang R, Wang J, Dong T et al (2019) Naringenin has a chemoprotective effect in MDA-MB-231 breast cancer cells via inhibition of caspase-3 and -9 activities. Oncol Lett 17:1217–1222. https://doi.org/10.3892/ol.2018.9704
Zadra G, Batista JL, Loda M (2015) Dissecting the dual role of AMPK in cancer: from experimental to human studies. Mol Cancer Res 13:1059–1072. https://doi.org/10.1158/1541-7786.MCR-15-0068
Zhang F, Dong W, Zeng W et al (2016) Naringenin prevents TGF-β1 secretion from breast cancer and suppresses pulmonary metastasis by inhibiting PKC activation. Breast Cancer Res 18:1–16. https://doi.org/10.1186/S13058-016-0698-0
Zhang FY, Du GJ, Zhang L et al (2009) Naringenin enhances the anti-tumor effect of doxorubicin through selectively inhibiting the activity of multidrug resistance-associated proteins but not P-glycoprotein. Pharm Res 26:914–925. https://doi.org/10.1007/s11095-008-9793-y
Zhang Y, Zhang GL, Sun X et al (2018) Establishment of a murine breast tumor model by subcutaneous or orthotopic implantation. Oncol Lett 15:6233–6240. https://doi.org/10.3892/ol.2018.8113
Zhao Z, Jin G, Ge Y, Guo Z (2019) Naringenin inhibits migration of breast cancer cells via inflammatory and apoptosis cell signaling pathways. Inflammopharmacology 27:1021–1036. https://doi.org/10.1007/s10787-018-00556-3
Acknowledgements
Authors acknowledge the Sun Pharmaceutical Industries Ltd. Vadodara for providing liposomal doxorubicin and animals required for this study. Authors also acknowledge Dr. S. K. Shah, Principal, Sardar Patel college of Pharmacy, Bakrol, Gujarat for providing animal usage approval for this project.
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The authors declare that all data were generated in-house and that no paper mill was used. Bharat Pateliya (BP): designed and performed the experiments, contributed to the data analysis and writing original draft. Vinod Burade (VB): reviewed and performed supervision. Sunita Goswami (SG): reviewed, results interpretation and work supervision. All the authors read and approved the final manuscript.
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This article does not contain any studies with human participants performed by any of the authors. All animal studies were carried out with prior approval from the Institutional Animal Ethics Committee (IAEC) and care of animals complied according to the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) guidelines, Government of India.
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Pateliya, B., Burade, V. & Goswami, S. Enhanced antitumor activity of doxorubicin by naringenin and metformin in breast carcinoma: an experimental study. Naunyn-Schmiedeberg's Arch Pharmacol 394, 1949–1961 (2021). https://doi.org/10.1007/s00210-021-02104-3
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DOI: https://doi.org/10.1007/s00210-021-02104-3