Research Article
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Synergistic Antiproliferative Impact of Capsaicin and Leflunomide in Murine Lung Cancer Cells

Year 2025, Volume: 44 Issue: 1, 35 - 41, 29.07.2025
https://doi.org/10.30782/jrvm.1671365

Abstract

Cancer remains a major global health issue, with resistance to conventional therapies driving the need for novel treatment strategies. This study investigated the synergistic effects of capsaicin (CAP), a natural bioactive compound, and Leflunomide (LEF), an immunosuppressive drug, in a murine lung cancer cell line. Capsaicin activates TRPV1 receptors, increasing intracellular calcium levels, while Leflunomide inhibits Dihydroorotate Dehydrogenase (DHODH), disrupting pyrimidine biosynthesis. Although neither agent exhibited significant cytotoxicity at low doses when applied individually, their combination resulted in a synergistic induction of cell death. These findings suggest that the combination of capsaicin and Leflunomide effectively limits cancer cell adaptation through metabolic and oxidative stress pathways, offering a promising anticancer mechanism. The results highlight the potential of this combination therapy for aggressive tumors such as lung cancer, emphasizing the need for further preclinical and clinical investigations.

References

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  • Miller KD, Nogueira L, Mariotto AB, et al. Cancer treatment and survivorship statistics, 2019. CA Cancer J Clin. 2019;69(5):363-385. doi:10.3322/caac.21565
  • Araghi M, Mannani R, Heidarnejad Maleki A, et al. Recent advances in non-small cell lung cancer targeted therapy; an update review. Cancer Cell Int. 2023;23(1):162. Published 2023 Aug 11. doi:10.1186/s12935-023-02990-y.
  • Webb JD, Simon MC. Novel insights into the molecular origins and treatment of lung cancer. Cell Cycle. 2010;9(20):4098-4105. doi:10.4161/cc.9.20.13588
  • Friedman JR, Nolan NA, Brown KC, et al. Anticancer Activity of Natural and Synthetic Capsaicin Analogs. J Pharmacol Exp Ther. 2018;364(3):462-473. doi:10.1124/jpet.117.243691
  • Vel SK, Ramakrishnan A, Sindya J, Rajanathadurai J, Perumal E. Evaluation of Cytotoxic and Anti-cancer Potential of Capsaicin on Lung Cancer Cell Line: An In Vitro Investigation. Cureus. 2024;16(8):e68119. Published 2024 Aug 29. doi:10.7759/cureus.68119
  • Hacıoğlu C. Kapsaisin U87 insan glioblastoma hücrelerinde epitelyal-mezenkimal geçişi regüle ederek hücre canlılığını baskılar. Sağlık Bilimleri Dergisi. 2024;33(2):271-278.
  • Chen YS, Lian YZ, Chen WC, et al. Lycium barbarum Polysaccharides and Capsaicin Inhibit Oxidative Stress, Inflammatory Responses, and Pain Signaling in Rats with Dextran Sulfate Sodium-Induced Colitis. Int J Mol Sci. 2022;23(5):2423. Published 2022 Feb 22. doi:10.3390/ijms23052423
  • Szallasi A. Capsaicin and cancer: Guilty as charged or innocent until proven guilty?. Temperature (Austin). 2022;10(1):35-49. Published 2022 Jan 11. doi:10.1080/23328940.2021.2017735Friedman JR, Wang T, Zhang L, et al. The role of capsaisin in promoting apoptosis in cancer therapy. Mol Nutr Food Res. 2017;61(9):1600762.
  • Liu T, Wang G, Tao H, et al. Capsaicin mediates caspases activation and induces apoptosis through P38 and JNK MAPK pathways in human renal carcinoma. BMC Cancer. 2016;16(1):790. Published 2016 Oct 12. doi:10.1186/s12885-016-2831-y
  • Sharma SK, Vij AS, Sharma M. Mechanisms and clinical uses of capsaicin. Eur J Pharmacol. 2013;720(1-3):55-62. doi:10.1016/j.ejphar.2013.10.053
  • Zhang C, Chu M. Leflunomide: A promising drug with good antitumor potential. Biochem Biophys Res Commun. 2018;496(2):726-730. doi:10.1016/j.bbrc.2018.01.107
  • Mirzapoiazova T, Tseng L, Mambetsariev B, et al. Teriflunomide/leflunomide synergizes with chemotherapeutics by decreasing mitochondrial fragmentation via DRP1 in SCLC. iScience. 2024;27(6):110132. Published 2024 May 27. doi:10.1016/j.isci.2024.110132
  • Xu X, Sun J, Song R, et al. Inhibition of p70 S6 kinase (S6K1) activity by A77 1726, the active metabolite of leflunomide, induces autophagy through TAK1-mediated AMPK and JNK activation. Oncotarget. 2017;8(18):30438-30454. doi:10.18632/oncotarget.16737.
  • Jiang X, Wang W, Lei L, et al. Antirheumatic drug leflunomide attenuates atherosclerosis by regulating lipid metabolism and endothelial dysfunction via DHODH/AMPK signaling pathway. Int J Biol Sci. 2024;20(10):3725-3741. Published 2024 Jul 2. doi:10.7150/ijbs.93465.
  • Wu J, Li Y, He Q, Yang X. Exploration of the Use of Natural Compounds in Combination with Chemotherapy Drugs for Tumor Treatment. Molecules. 2023;28(3):1022. Published 2023 Jan 19. doi:10.3390/molecules28031022.
  • Orellana EA, Kasinski AL. Sulforhodamine B (SRB) Assay in Cell Culture to Investigate Cell Proliferation. Bio Protoc. 2016;6(21):e1984. doi:10.21769/BioProtoc.1984.
  • Han TH, Park MK, Nakamura H, Ban HS. Capsaicin inhibits HIF-1α accumulation through suppression of mitochondrial respiration in lung cancer cells. Biomed Pharmacother. 2022;146:112500. doi:10.1016/j.biopha.2021.112500.
  • Jiang L, Zhang W, Li W, Ling C, Jiang M. Anti-inflammatory drug, leflunomide and its metabolite teriflunomide inhibit NSCLC proliferation in vivo and in vitro. Toxicol Lett. 2018;282:154-165. doi:10.1016/j.toxlet.2017.10.013.
  • Shoichet BK. Interpreting steep dose-response curves in early inhibitor discovery. J Med Chem. 2006;49(25):7274-7277. doi:10.1021/jm061103g.
  • Ianevski A, He L, Aittokallio T, Tang J. SynergyFinder: a web application for analyzing drug combination dose-response matrix data [published correction appears in Bioinformatics. 2020 Apr 15;36(8):2645. doi: 10.1093/bioinformatics/btaa102.]. Bioinformatics. 2017;33(15):2413-2415. doi:10.1093/bioinformatics/btx162.
  • Franken NA, Rodermond HM, Stap J, Haveman J, van Bree C. Clonogenic assay of cells in vitro. Nat Protoc. 2006;1(5):2315-2319. doi:10.1038/nprot.2006.339.
  • Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 2012;9(7):671-675. doi:10.1038/nmeth.2089.
  • Roviello G, Nobili S, Mini E. A potential novel drug combination for adjuvant therapy in lung cancer: reflections on an early phase of clinical development. J Chemother. 2019;31(6):295-296. doi:10.1080/1120009X.2019.1651014.
  • Zhao E, Jiang X, Cui H. Bombyx mori Dihydroorotate Dehydrogenase: Knockdown Inhibits Cell Growth and Proliferation via Inducing Cell Cycle Arrest. Int J Mol Sci. 2018;19(9):2581. Published 2018 Aug 30. doi:10.3390/ijms19092581.
  • Fang J, Uchiumi T, Yagi M, et al. Dihydro-orotate dehydrogenase is physically associated with the respiratory complex and its loss leads to mitochondrial dysfunction. Biosci Rep. 2013;33(2):e00021. Published 2013 Feb 5. doi:10.1042/BSR20120097.
  • Yang F, Zheng J. Understand spiciness: mechanism of TRPV1 channel activation by capsaicin. Protein Cell. 2017;8(3):169-177. doi:10.1007/s13238-016-0353-7.
  • Knoell J, Chillappagari S, Knudsen L, et al. PACS2-TRPV1 axis is required for ER-mitochondrial tethering during ER stress and lung fibrosis. Cell Mol Life Sci. 2022;79(3):151. Published 2022 Feb 25. doi:10.1007/s00018-022-04189-2.
  • Ladds MJGW, van Leeuwen IMM, Drummond CJ, et al. Publisher Correction: A DHODH inhibitor increases p53 synthesis and enhances tumor cell killing by p53 degradation blockage. Nat Commun. 2023;14(1):5019. Published 2023 Aug 18. doi:10.1038/s41467-023-40764-2.
  • Wu Q, Wu W, Fu B, Shi L, Wang X, Kuca K. JNK signaling in cancer cell survival. Med Res Rev. 2019;39(6):2082-2104. doi:10.1002/med.21574.
  • Jia XY, Yang Y, Jia XT, et al. Capsaicin pretreatment attenuates salt-sensitive hypertension by alleviating AMPK/Akt/Nrf2 pathway in hypothalamic paraventricular nucleus. Front Neurosci. 2024;18:1416522. Published 2024 May 30. doi:10.3389/fnins.2024.1416522.
  • Merritt JC, Richbart SD, Moles EG, et al. Anti-cancer activity of sustained release capsaicin formulations. Pharmacol Ther. 2022;238:108177. doi:10.1016/j.pharmthera.2022.108177.
  • Qi C, Wang D, Gong X, et al. Co-Delivery of Curcumin and Capsaicin by Dual-Targeting Liposomes for Inhibition of aHSC-Induced Drug Resistance and Metastasis. ACS Appl Mater Interfaces. 2021;13(14):16019-16035. doi:10.1021/acsami.0c23137.
  • Krizanova O, Steliarova I, Csaderova L, Pastorek M, Hudecova S. Capsaicin induces apoptosis in PC12 cells through ER stress. Oncol Rep. 2014;31(2):581-588. doi:10.3892/or.2013.2921.
  • Koundinya M, Sudhalter J, Courjaud A, et al. Dependence on the Pyrimidine Biosynthetic Enzyme DHODH Is a Synthetic Lethal Vulnerability in Mutant KRAS-Driven Cancers. Cell Chem Biol. 2018;25(6):705-717.e11. doi:10.1016/j.chembiol.2018.03.005.

Year 2025, Volume: 44 Issue: 1, 35 - 41, 29.07.2025
https://doi.org/10.30782/jrvm.1671365

Abstract

References

  • Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020. CA Cancer J Clin. 2020;70(1):7-30.
  • Miller KD, Nogueira L, Mariotto AB, et al. Cancer treatment and survivorship statistics, 2019. CA Cancer J Clin. 2019;69(5):363-385. doi:10.3322/caac.21565
  • Araghi M, Mannani R, Heidarnejad Maleki A, et al. Recent advances in non-small cell lung cancer targeted therapy; an update review. Cancer Cell Int. 2023;23(1):162. Published 2023 Aug 11. doi:10.1186/s12935-023-02990-y.
  • Webb JD, Simon MC. Novel insights into the molecular origins and treatment of lung cancer. Cell Cycle. 2010;9(20):4098-4105. doi:10.4161/cc.9.20.13588
  • Friedman JR, Nolan NA, Brown KC, et al. Anticancer Activity of Natural and Synthetic Capsaicin Analogs. J Pharmacol Exp Ther. 2018;364(3):462-473. doi:10.1124/jpet.117.243691
  • Vel SK, Ramakrishnan A, Sindya J, Rajanathadurai J, Perumal E. Evaluation of Cytotoxic and Anti-cancer Potential of Capsaicin on Lung Cancer Cell Line: An In Vitro Investigation. Cureus. 2024;16(8):e68119. Published 2024 Aug 29. doi:10.7759/cureus.68119
  • Hacıoğlu C. Kapsaisin U87 insan glioblastoma hücrelerinde epitelyal-mezenkimal geçişi regüle ederek hücre canlılığını baskılar. Sağlık Bilimleri Dergisi. 2024;33(2):271-278.
  • Chen YS, Lian YZ, Chen WC, et al. Lycium barbarum Polysaccharides and Capsaicin Inhibit Oxidative Stress, Inflammatory Responses, and Pain Signaling in Rats with Dextran Sulfate Sodium-Induced Colitis. Int J Mol Sci. 2022;23(5):2423. Published 2022 Feb 22. doi:10.3390/ijms23052423
  • Szallasi A. Capsaicin and cancer: Guilty as charged or innocent until proven guilty?. Temperature (Austin). 2022;10(1):35-49. Published 2022 Jan 11. doi:10.1080/23328940.2021.2017735Friedman JR, Wang T, Zhang L, et al. The role of capsaisin in promoting apoptosis in cancer therapy. Mol Nutr Food Res. 2017;61(9):1600762.
  • Liu T, Wang G, Tao H, et al. Capsaicin mediates caspases activation and induces apoptosis through P38 and JNK MAPK pathways in human renal carcinoma. BMC Cancer. 2016;16(1):790. Published 2016 Oct 12. doi:10.1186/s12885-016-2831-y
  • Sharma SK, Vij AS, Sharma M. Mechanisms and clinical uses of capsaicin. Eur J Pharmacol. 2013;720(1-3):55-62. doi:10.1016/j.ejphar.2013.10.053
  • Zhang C, Chu M. Leflunomide: A promising drug with good antitumor potential. Biochem Biophys Res Commun. 2018;496(2):726-730. doi:10.1016/j.bbrc.2018.01.107
  • Mirzapoiazova T, Tseng L, Mambetsariev B, et al. Teriflunomide/leflunomide synergizes with chemotherapeutics by decreasing mitochondrial fragmentation via DRP1 in SCLC. iScience. 2024;27(6):110132. Published 2024 May 27. doi:10.1016/j.isci.2024.110132
  • Xu X, Sun J, Song R, et al. Inhibition of p70 S6 kinase (S6K1) activity by A77 1726, the active metabolite of leflunomide, induces autophagy through TAK1-mediated AMPK and JNK activation. Oncotarget. 2017;8(18):30438-30454. doi:10.18632/oncotarget.16737.
  • Jiang X, Wang W, Lei L, et al. Antirheumatic drug leflunomide attenuates atherosclerosis by regulating lipid metabolism and endothelial dysfunction via DHODH/AMPK signaling pathway. Int J Biol Sci. 2024;20(10):3725-3741. Published 2024 Jul 2. doi:10.7150/ijbs.93465.
  • Wu J, Li Y, He Q, Yang X. Exploration of the Use of Natural Compounds in Combination with Chemotherapy Drugs for Tumor Treatment. Molecules. 2023;28(3):1022. Published 2023 Jan 19. doi:10.3390/molecules28031022.
  • Orellana EA, Kasinski AL. Sulforhodamine B (SRB) Assay in Cell Culture to Investigate Cell Proliferation. Bio Protoc. 2016;6(21):e1984. doi:10.21769/BioProtoc.1984.
  • Han TH, Park MK, Nakamura H, Ban HS. Capsaicin inhibits HIF-1α accumulation through suppression of mitochondrial respiration in lung cancer cells. Biomed Pharmacother. 2022;146:112500. doi:10.1016/j.biopha.2021.112500.
  • Jiang L, Zhang W, Li W, Ling C, Jiang M. Anti-inflammatory drug, leflunomide and its metabolite teriflunomide inhibit NSCLC proliferation in vivo and in vitro. Toxicol Lett. 2018;282:154-165. doi:10.1016/j.toxlet.2017.10.013.
  • Shoichet BK. Interpreting steep dose-response curves in early inhibitor discovery. J Med Chem. 2006;49(25):7274-7277. doi:10.1021/jm061103g.
  • Ianevski A, He L, Aittokallio T, Tang J. SynergyFinder: a web application for analyzing drug combination dose-response matrix data [published correction appears in Bioinformatics. 2020 Apr 15;36(8):2645. doi: 10.1093/bioinformatics/btaa102.]. Bioinformatics. 2017;33(15):2413-2415. doi:10.1093/bioinformatics/btx162.
  • Franken NA, Rodermond HM, Stap J, Haveman J, van Bree C. Clonogenic assay of cells in vitro. Nat Protoc. 2006;1(5):2315-2319. doi:10.1038/nprot.2006.339.
  • Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 2012;9(7):671-675. doi:10.1038/nmeth.2089.
  • Roviello G, Nobili S, Mini E. A potential novel drug combination for adjuvant therapy in lung cancer: reflections on an early phase of clinical development. J Chemother. 2019;31(6):295-296. doi:10.1080/1120009X.2019.1651014.
  • Zhao E, Jiang X, Cui H. Bombyx mori Dihydroorotate Dehydrogenase: Knockdown Inhibits Cell Growth and Proliferation via Inducing Cell Cycle Arrest. Int J Mol Sci. 2018;19(9):2581. Published 2018 Aug 30. doi:10.3390/ijms19092581.
  • Fang J, Uchiumi T, Yagi M, et al. Dihydro-orotate dehydrogenase is physically associated with the respiratory complex and its loss leads to mitochondrial dysfunction. Biosci Rep. 2013;33(2):e00021. Published 2013 Feb 5. doi:10.1042/BSR20120097.
  • Yang F, Zheng J. Understand spiciness: mechanism of TRPV1 channel activation by capsaicin. Protein Cell. 2017;8(3):169-177. doi:10.1007/s13238-016-0353-7.
  • Knoell J, Chillappagari S, Knudsen L, et al. PACS2-TRPV1 axis is required for ER-mitochondrial tethering during ER stress and lung fibrosis. Cell Mol Life Sci. 2022;79(3):151. Published 2022 Feb 25. doi:10.1007/s00018-022-04189-2.
  • Ladds MJGW, van Leeuwen IMM, Drummond CJ, et al. Publisher Correction: A DHODH inhibitor increases p53 synthesis and enhances tumor cell killing by p53 degradation blockage. Nat Commun. 2023;14(1):5019. Published 2023 Aug 18. doi:10.1038/s41467-023-40764-2.
  • Wu Q, Wu W, Fu B, Shi L, Wang X, Kuca K. JNK signaling in cancer cell survival. Med Res Rev. 2019;39(6):2082-2104. doi:10.1002/med.21574.
  • Jia XY, Yang Y, Jia XT, et al. Capsaicin pretreatment attenuates salt-sensitive hypertension by alleviating AMPK/Akt/Nrf2 pathway in hypothalamic paraventricular nucleus. Front Neurosci. 2024;18:1416522. Published 2024 May 30. doi:10.3389/fnins.2024.1416522.
  • Merritt JC, Richbart SD, Moles EG, et al. Anti-cancer activity of sustained release capsaicin formulations. Pharmacol Ther. 2022;238:108177. doi:10.1016/j.pharmthera.2022.108177.
  • Qi C, Wang D, Gong X, et al. Co-Delivery of Curcumin and Capsaicin by Dual-Targeting Liposomes for Inhibition of aHSC-Induced Drug Resistance and Metastasis. ACS Appl Mater Interfaces. 2021;13(14):16019-16035. doi:10.1021/acsami.0c23137.
  • Krizanova O, Steliarova I, Csaderova L, Pastorek M, Hudecova S. Capsaicin induces apoptosis in PC12 cells through ER stress. Oncol Rep. 2014;31(2):581-588. doi:10.3892/or.2013.2921.
  • Koundinya M, Sudhalter J, Courjaud A, et al. Dependence on the Pyrimidine Biosynthetic Enzyme DHODH Is a Synthetic Lethal Vulnerability in Mutant KRAS-Driven Cancers. Cell Chem Biol. 2018;25(6):705-717.e11. doi:10.1016/j.chembiol.2018.03.005.
There are 35 citations in total.

Details

Primary Language English
Subjects Veterinary Sciences (Other)
Journal Section Research Article
Authors

Aybike Sarıoğlu Bozkurt 0000-0002-8287-6617

Submission Date April 7, 2025
Acceptance Date May 13, 2025
Publication Date July 29, 2025
Published in Issue Year 2025 Volume: 44 Issue: 1

Cite

APA Sarıoğlu Bozkurt, A. (2025). Synergistic Antiproliferative Impact of Capsaicin and Leflunomide in Murine Lung Cancer Cells. Journal of Research in Veterinary Medicine, 44(1), 35-41. https://doi.org/10.30782/jrvm.1671365
AMA Sarıoğlu Bozkurt A. Synergistic Antiproliferative Impact of Capsaicin and Leflunomide in Murine Lung Cancer Cells. J Res Vet Med. July 2025;44(1):35-41. doi:10.30782/jrvm.1671365
Chicago Sarıoğlu Bozkurt, Aybike. “Synergistic Antiproliferative Impact of Capsaicin and Leflunomide in Murine Lung Cancer Cells”. Journal of Research in Veterinary Medicine 44, no. 1 (July 2025): 35-41. https://doi.org/10.30782/jrvm.1671365.
EndNote Sarıoğlu Bozkurt A (July 1, 2025) Synergistic Antiproliferative Impact of Capsaicin and Leflunomide in Murine Lung Cancer Cells. Journal of Research in Veterinary Medicine 44 1 35–41.
IEEE A. Sarıoğlu Bozkurt, “Synergistic Antiproliferative Impact of Capsaicin and Leflunomide in Murine Lung Cancer Cells”, J Res Vet Med, vol. 44, no. 1, pp. 35–41, 2025, doi: 10.30782/jrvm.1671365.
ISNAD Sarıoğlu Bozkurt, Aybike. “Synergistic Antiproliferative Impact of Capsaicin and Leflunomide in Murine Lung Cancer Cells”. Journal of Research in Veterinary Medicine 44/1 (July2025), 35-41. https://doi.org/10.30782/jrvm.1671365.
JAMA Sarıoğlu Bozkurt A. Synergistic Antiproliferative Impact of Capsaicin and Leflunomide in Murine Lung Cancer Cells. J Res Vet Med. 2025;44:35–41.
MLA Sarıoğlu Bozkurt, Aybike. “Synergistic Antiproliferative Impact of Capsaicin and Leflunomide in Murine Lung Cancer Cells”. Journal of Research in Veterinary Medicine, vol. 44, no. 1, 2025, pp. 35-41, doi:10.30782/jrvm.1671365.
Vancouver Sarıoğlu Bozkurt A. Synergistic Antiproliferative Impact of Capsaicin and Leflunomide in Murine Lung Cancer Cells. J Res Vet Med. 2025;44(1):35-41.