Rasagiline Inhibits Human Melanoma Cell Viability and Interacts Synergistically with Mitoxantrone and Antagonistically with Cisplatin—In Vitro Isobolographic Studies
Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Drugs
2.3. MTT Assay
2.4. LDH Test
2.5. BrdU Test
2.6. Flow Cytometry—Cell-Cycle Analysis
2.7. Isobolographic Analysis
2.8. Statistical Analysis
3. Results
3.1. Effects of Rasagiline on Malignant Melanoma Cell Viability in the MTT Assay
3.2. Effects of Rasagiline on Malignant Melanoma Cell Cytotoxicity in the LDH Assay
3.3. Effects of Rasagiline on Malignant Melanoma Cell Proliferation in the BrdU Assay
3.4. Effects of Rasagiline on Malignant Melanoma Cell Cycle Progression
3.5. Isobolographic Interactions Between Rasagiline and Cisplatin and Mitoxantrone
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Abbreviations
CDDP | cisplatin |
MTX | mitoxantrone |
IC50 | median inhibitory concentration |
PD | Parkinson’s disease |
MC1R | melanocortin 1 receptor |
CYP2D6 | cytochrome P450 2D6 |
VDR | vitamin D receptor |
L-DOPA | levodopa |
UV | ultraviolet |
MAO-B | monoamine oxidase subtype B |
ECACC | European Collection of Cell Cultures |
ATCC | American Type Culture Collection |
IAN | indole-3-acetonitrile |
4-S-CAP | 4-S-cysteaminylphenol |
References
- Samii, A.; Nutt, J.G.; Ransom, B.R. Parkinson’s disease. Lancet 2004, 363, 1783–1793. [Google Scholar] [CrossRef]
- Lesage, S.; Brice, A. Parkinson’s disease: From monogenic forms to genetic susceptibility factors. Hum. Mol. Genet. 2009, 18, R48–R59. [Google Scholar] [CrossRef]
- Kendal, W.S. Dying with cancer: The influence of age, comorbidity, and cancer site. Cancer 2008, 112, 1354–1362. [Google Scholar] [CrossRef]
- Moller, H.; Mellemkjaer, L.; McLaughlin, J.K.; Olsen, J.H. Occurrence of different cancers in patients with Parkinson’s disease. BMJ 1995, 310, 1500–1501. [Google Scholar] [CrossRef]
- Olsen, J.H.; Friis, S.; Frederiksen, K.; McLaughlin, J.K.; Mellemkjaer, L.; Møller, H. Atypical cancer pattern in patients with Parkinson’s disease. Br. J. Cancer 2005, 92, 201–205. [Google Scholar] [CrossRef]
- Gao, X.; Simon, K.C.; Han, J.; Schwarzschild, M.A.; Ascherio, A. Family history of melanoma and Parkinson disease risk. Neurology 2009, 73, 1286–1291. [Google Scholar] [CrossRef]
- Huang, P.; Yang, X.-D.; Chen, S.-D.; Xiao, Q. The association between Parkinson’s disease and melanoma: A systematic review and meta-analysis. Transl. Neurodegener. 2015, 4, 21. [Google Scholar] [CrossRef] [PubMed]
- Zhang, X.; Guarin, D.; Mohammadzadehhonarvar, N.; Chen, X.; Gao, X. Parkinson’s disease and cancer: A systematic review and meta-analysis of over 17 million participants. BMJ Open 2021, 11, e046329. [Google Scholar] [CrossRef] [PubMed]
- Ferreira, J.J.; Neutel, D.; Mestre, T.; Coelho, M.; Rosa, M.M.; Rascol, O.; Sampaio, C. Skin cancer and Parkinson’s disease. Mov. Disord. 2010, 25, 139–148. [Google Scholar] [CrossRef]
- Pan, T.; Li, X.; Jankovic, J. The association between Parkinson’s disease and melanoma. Int. J. Cancer 2011, 128, 2251–2260. [Google Scholar] [CrossRef] [PubMed]
- Bose, A.; Petsko, G.A.; Eliezer, D. Parkinson’s disease and melanoma: Co-occurrence and mechanisms. J. Park. Dis. 2018, 8, 385–398. [Google Scholar] [CrossRef]
- Chen, X.; Feng, D.; Schwarzschild, M.A.; Gao, X. Red hair, MC1R variants, and risk for Parkinson’s disease—A meta-analysis. Ann. Clin. Transl. Neurol. 2017, 4, 212–216. [Google Scholar] [CrossRef]
- Gao, X.; Simon, K.C.; Han, J.; Schwarzschild, M.A.; Ascherio, A. Genetic determinants of hair color and Parkinson’s disease risk. Ann. Neurol. 2009, 65, 76–82. [Google Scholar] [CrossRef]
- Fullard, M.E.; Duda, J.E. A Review of the relationship between vitamin D and Parkinson disease symptoms. Front. Neurol. 2020, 11, 454. [Google Scholar] [CrossRef]
- Siple, J.F.; Schneider, D.C.; Wanlass, W.A.; Rosenblatt, B.K. Levodopa therapy and the risk of malignant melanoma. Ann. Pharmacother. 2000, 34, 382–385. [Google Scholar] [CrossRef] [PubMed]
- Weissenrieder, J.S.; Neighbors, J.D.; Mailman, R.B.; Hohl, R.J. Cancer and the Dopamine D2Receptor: A Pharmacological Perspective. J. Pharmacol. Exp. Ther. 2019, 370, 111–126. [Google Scholar] [CrossRef]
- Skibba, J.L.; Pinckley, J.; Gilbert, E.F.; Johnson, R.O. Multiple primary melanoma following administration of levodopa. Arch. Pathol. 1972, 93, 556–561. [Google Scholar]
- Weiner, W.J.; Singer, C.; Sanchez-Ramos, J.R.; Goldenberg, J.N. Levodopa, melanoma, and Parkinson’s disease. Neurology 1993, 43, 674–677. [Google Scholar] [CrossRef] [PubMed]
- Whiteman, D.C.; Stickley, M.; Watt, P.; Hughes, M.C.; Davis, M.B.; Green, A.C. Anatomic site, sun exposure, and risk of cutaneous melanoma. J. Clin. Oncol. 2006, 24, 3172–3177. [Google Scholar] [CrossRef]
- Ivry, G.B.; Ogle, C.A.; Shim, E.K. Role of sun exposure in melanoma. Dermatol. Surg. 2006, 32, 481–492. [Google Scholar] [CrossRef] [PubMed]
- Lew, M.F.; Hauser, R.A.; Hurtig, H.I.; Ondo, W.G.; Wojcieszek, J.; Goren, T.; Fitzer-Attas, C.J. Long-term efficacy of rasagiline in early Parkinson’s disease. Int. J. Neurosci. 2010, 120, 404–408. [Google Scholar] [CrossRef]
- Bar-Am, O.; Weinreb, O.; Amit, T.; Youdim, M.B.H. The neuroprotective mechanism of 1-®-aminoindan, the major metabolite of the anti-parkinsonian drug rasagiline. J. Neurochem. 2010, 112, 1131–1137. [Google Scholar] [CrossRef]
- Rabey, J.M.; Sagi, I.; Huberman, M.; Melamed, E.; Korczyn, A.; Giladi, N.; Inzelberg, R.; Djaldetti, R.; Klein, C.; Berecz, G.; et al. Rasagiline mesylate, a new MAO-B inhibitor for the treatment of Parkinson’s disease: A double-blind study as adjunctive therapy to levodopa. Clin. Neuropharmacol. 2000, 23, 324–330. [Google Scholar] [CrossRef]
- Weinreb, O.; Amit, T.; Bar-Am, O.; Youdim, M.B. Rasagiline: A novel anti-Parkinsonian monoamine oxidase-B inhibitor with neuroprotective activity. Prog. Neurobiol. 2010, 92, 330–344. [Google Scholar] [CrossRef]
- Parkinson Study Group. A randomized placebo-controlled trial of rasagiline in levodopa-treated patients with Parkinson disease and motor fluctuations: The PRESTO study. Arch. Neurol. 2005, 62, 241–248. [Google Scholar] [CrossRef]
- Parkinson Study Group. A controlled trial of rasagiline in early Parkinson disease: The TEMPO study. Arch. Neurol. 2002, 59, 1937–1943. [Google Scholar] [CrossRef]
- Teva Pharmaceutical Industries Ltd. Azilect® Package Insert; Teva Pharmaceutical Industries Ltd.: Kfar Saba, Israel, 2006. [Google Scholar]
- Johannes, C.B.; Saltus, C.W.; Kaye, J.A.; Calingaert, B.; Kaplan, S.; Gordon, M.F.; Andrews, E.B. The risk of melanoma with rasagiline compared with other antiparkinsonian medications: A retrospective cohort study in the United States medicare database. Pharmacoepidemiol. Drug Saf. 2022, 31, 643–651. [Google Scholar] [CrossRef] [PubMed]
- Krasowska, D.; Gerkowicz, A.; Wróblewska-Łuczka, P.; Grabarska, A.; Załuska-Ogryzek, K.; Krasowska, D.; Łuszczki, J.J. Anticancer Activity of Amantadine and Evaluation of Its Interactions with Selected Cytostatics in Relation to Human Melanoma Cells. Int. J. Mol. Sci. 2022, 23, 7653. [Google Scholar] [CrossRef] [PubMed]
- Marzęda, P.; Wróblewska-Łuczka, P.; Drozd, M.; Florek-Łuszczki, M.; Załuska-Ogryzek, K.; Łuszczki, J.J. Cannabidiol Interacts Antagonistically with Cisplatin and Additively with Mitoxantrone in Various Melanoma Cell Lines—An Isobolographic Analysis. Int. J. Mol. Sci. 2022, 23, 6752. [Google Scholar] [CrossRef]
- Wróblewska-Łuczka, P.; Kulenty, L.; Załuska-Ogryzek, K.; Góralczyk, A.; Łuszczki, J.J. Screening of the Antimelanoma Activity of Monoterpenes—In Vitro Experiments on Four Human Melanoma Lines. Curr. Issues Mol. Biol. 2025, 47, 97. [Google Scholar] [CrossRef] [PubMed]
- Kaja, S.; Payne, A.J.; Singh, T.; Ghuman, J.K.; Sieck, E.G.; Koulen, P. An optimized lactate dehydrogenase release assay for screening of drug candidates in neuroscience. J. Pharmacol. Toxicol. Methods 2015, 73, 1–6. [Google Scholar] [CrossRef] [PubMed]
- Litchfield, J.T., Jr.; Wilcoxon, F. A Simplified Method of Evaluating Dose-Effect Experiments. J. Pharmacol. Exp. Ther. 1949, 96, 99–113. [Google Scholar] [CrossRef] [PubMed]
- Luszczki, J.J. Isobolographic analysis of interaction between drugs with nonparallel dose–response relationship curves: A practical application. Naunyn-Schmiedeberg’s Arch. Pharmacol. 2007, 375, 105–114. [Google Scholar] [CrossRef] [PubMed]
- Loewe, S. The problem of synergism and antagonism of combined drugs. Arzneimittelforschung 1953, 3, 285–290. [Google Scholar]
- Tallarida, R.J. Quantitative methods for assessing drug synergism. Genes Cancer 2011, 2, 1003–1008. [Google Scholar] [CrossRef]
- Huang, R.-Y.; Pei, L.; Liu, Q.; Chen, S.; Dou, H.; Shu, G.; Yuan, Z.-X.; Lin, J.; Peng, G.; Zhang, W.; et al. Isobologram Analysis: A Comprehensive Review of Methodology and Current Research. Front. Pharmacol. 2019, 10, 1222. [Google Scholar] [CrossRef]
- Duarte, D.; Amaro, F.; Silva, I.; Silva, D.; Fresco, P.; Oliveira, J.C.; Reguengo, H.; Gonçalves, J.; Vale, N. Carbidopa Alters Tryptophan Metabolism in Breast Cancer and Melanoma Cells Leading to the Formation of Indole-3-Acetonitrile, a Pro-Proliferative Metabolite. Biomolecules 2019, 9, 409. [Google Scholar] [CrossRef]
- Ogura, J.; Miyauchi, S.; Shimono, K.; Yang, S.; Gonchigar, S.; Ganapathy, V.; Bhutia, Y.D. Carbidopa is an activator of aryl hydrocarbon receptor with potential for cancer therapy. Biochem. J. 2017, 474, 3391–3402. [Google Scholar] [CrossRef]
- Thomas, C.; Wafa, L.A.; Lamoureux, F.; Cheng, H.; Fazli, L.; Gleave, M.E.; Rennie, P.S. Carbidopa enhances antitumoral activity of bicalutamide on the androgen receptor-axis in castration-resistant prostate tumors. Prostate 2012, 72, 875–885. [Google Scholar] [CrossRef]
- Gąsowska-Bajger, B.; Frąckowiak-Wojtasek, B.; Koj, S.; Cichoń, T.; Smolarczyk, R.; Szala, S.; Wojtasek, H. Oxidation of carbidopa by tyrosinase and its effect on murine melanoma. Bioorganic Med. Chem. Lett. 2009, 19, 3507–3510. [Google Scholar] [CrossRef]
- Miura, T.; Jimbow, K.; Ito, S. The in vivo antimelanoma effect of 4-S-cysteaminylphenol and its N-acetyl derivative. Int. J. Cancer 1990, 46, 931–934. [Google Scholar] [CrossRef]
- Gurney, H.; Coates, A.; Kefford, R. The use of L-DOPA and carbidopa in metastatic malignant melanoma. J. Investig. Dermatol. 1991, 96, 85–87. [Google Scholar] [CrossRef] [PubMed]
- Tazik, S.; Johnson, S.; Lu, D.; Johnson, C.; Youdim, M.B.H.; Stockmeier, C.A.; Ou, X.-M. Comparative neuroprotective effects of rasagiline and aminoindan with selegiline on dexamethasone-induced brain cell apoptosis. Neurotox. Res. 2009, 15, 284–290. [Google Scholar] [CrossRef] [PubMed]
- Akao, Y.; Maruyama, W.; Shimizu, S.; Yi, H.; Nakagawa, Y.; Shamoto-Nagai, M.; Youdim, M.B.H.; Tsujimoto, Y.; Naoi, M. Mitochondrial permeability transition mediates apoptosis induced by N-methyl(R)salsolinol, an endogenous neurotoxin, and is inhibited by Bcl-2 and rasagiline, N-propargyl-1(R)-aminoindan. J. Neurochem. 2002, 82, 913–923. [Google Scholar] [CrossRef] [PubMed]
- Naoi, M.; Maruyama, W.; Inaba-Hasegawa, K. Revelation in the neuroprotective functions of rasagiline and selegiline: The induction of distinct genes by different mechanisms. Expert Rev. Neurother. 2013, 13, 671–684. [Google Scholar] [CrossRef]
- Naoi, M.; Maruyama, W.; Yi, H. Rasagiline prevents apoptosis induced by PK11195, a ligand of the outer membrane translocator protein (18 kDa), in SH-SY5Y cells through suppression of cytochrome c release from mitochondria. J. Neural Transm. 2013, 120, 1539–1551. [Google Scholar] [CrossRef]
- Inaba-Hasegawa, K.; Shamoto-Nagai, M.; Maruyama, W.; Naoi, M. Type B and A monoamine oxidase and their inhibitors regulate the gene expression of Bcl-2 and neurotrophic factors in human glioblastoma U118MG cells: Different signaling pathways for neuroprotection by selegiline and rasagiline. J. Neural Transm. 2017, 124, 1055–1066. [Google Scholar] [CrossRef]
- Meier-Davis, S.R.; Dines, K.; Arjmand, F.M.; Hamlin, R.; Huang, B.; Wen, J.; Christianson, C.; Shudo, J.; Nagata, T. Comparison of oral and transdermal administration of rasagiline mesylate on human melanoma tumor growth in vivo. Cutan. Ocul. Toxicol. 2012, 31, 312–317. [Google Scholar] [CrossRef]
- Indrayanto, G.; Putra, G.S.; Suhud, F. Validation of in-vitro bioassay methods: Application in herbal drug research. Profiles Drug. Subst. Excip. Relat. Methodol. 2021, 46, 273–307. [Google Scholar] [CrossRef]
- Marzęda, P.; Wróblewska-Łuczka, P.; Florek-Łuszczki, M.; Góralczyk, A.; Łuszczki, J.J. AM1172 (a hydrolysis-resistant endocannabinoid analog that inhibits anandamide cellular uptake) reduces the viability of the various melanoma cells, but it exerts significant cytotoxic effects on healthy cells: An in vitro study based on isobolographic analysis. Pharmacol. Rep. 2024, 76, 154–170. [Google Scholar] [CrossRef]
Drug/Cell Line | FM55P | FM55M2 | A375 | SK-MEL28 | References |
---|---|---|---|---|---|
Rasagiline mesylate | 349.44 ± 97.52 µM | 117.45 ± 56.89 µM | 280.69 ± 53.30 µM | 402.89 ± 113.83 µM | this study |
Rasagiline mesylate | 93.42 ± 26.07 µg/mL | 31.4 ± 15.21 µg/mL | 75.04 ± 14.25 µg/mL | 107.71 ± 30.43 µg/mL | this study |
Cisplatin | 1.49 ± 0.30 μM | 1.70 ± 0.35 μM | 1.29 ± 0.34 μM | 3.30 ± 0.70 μM | [29,30] |
Mitoxantrone | 0.35 ± 0.10 μM | 0.13 ± 0.02 μM | 0.04 ± 0.02 μM | 1.74 ± 0.51 μM | [30] |
Drug Combination | Cell Line | IC50mix (nmix) [µg/mL] | IC50add (nadd) [µg/mL] | t Statistics | Interaction |
---|---|---|---|---|---|
Rasagiline+CDDP | A375 | 71.66 ± 12.37 * (96) | 37.71 ± 7.17 (212) | t161 = 2.374; p = 0.019 | Antagonistic |
FM55P | 85.15 ± 18.11 (96) | 46.98 ± 13.10 (188) | t193 = 1.708; p = 0.089 | Additive | |
SK-MEL28 | 105.10 ± 15.84 * (96) | 54.35 ± 15.34 (188) | t247 = 2.302; p = 0.022 | Antagonistic | |
Rasagiline+MTX | FM55P | 11.67 ± 3.28 * (96) | 46.80 ± 14.61 (164) | t179 = 2.346; p = 0.020 | Synergistic |
SK-MEL28 | 21.85 ± 13.08 (96) | 54.31 ± 17.16 (164) | t258 = 1.504; p = 0.134 | Additive |
Drug Combination | Cell Line | IC50mix (nmix) [µg/mL] | Lower IC50add (nadd) [µg/mL] | Upper IC50add (nadd) [µg/mL] | t Statistics | Interaction |
---|---|---|---|---|---|---|
Rasagiline+CDDP | FM55M2 | 32.24 ± 8.32 (96) | 11.22 ± 7.41 (182) | 20.57 ± 7.47 (182) | t231 = 1.044; p = 0.298 | Additive |
Rasagiline+MTX | FM55M2 | 4.06 ± 1.27 (96) | 8.75 ± 7.67 (182) | 22.64 ± 8.33 (182) | t191 = 0.6033; p = 0.547 | Additive |
A375 | 12.95 ± 8.29 (96) | 26.02 ± 13.18 (242) | 49.17 ± 15.33 (242) | t336 = 0.8394; p = 0.402 | Additive |
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Krasowska, D.; Wróblewska-Łuczka, P.; Chojnacki, M.; Załuska-Ogryzek, K.; Kurzepa, J.; Łuszczki, J.J. Rasagiline Inhibits Human Melanoma Cell Viability and Interacts Synergistically with Mitoxantrone and Antagonistically with Cisplatin—In Vitro Isobolographic Studies. Cancers 2025, 17, 2563. https://doi.org/10.3390/cancers17152563
Krasowska D, Wróblewska-Łuczka P, Chojnacki M, Załuska-Ogryzek K, Kurzepa J, Łuszczki JJ. Rasagiline Inhibits Human Melanoma Cell Viability and Interacts Synergistically with Mitoxantrone and Antagonistically with Cisplatin—In Vitro Isobolographic Studies. Cancers. 2025; 17(15):2563. https://doi.org/10.3390/cancers17152563
Chicago/Turabian StyleKrasowska, Danuta, Paula Wróblewska-Łuczka, Michał Chojnacki, Katarzyna Załuska-Ogryzek, Jacek Kurzepa, and Jarogniew J. Łuszczki. 2025. "Rasagiline Inhibits Human Melanoma Cell Viability and Interacts Synergistically with Mitoxantrone and Antagonistically with Cisplatin—In Vitro Isobolographic Studies" Cancers 17, no. 15: 2563. https://doi.org/10.3390/cancers17152563
APA StyleKrasowska, D., Wróblewska-Łuczka, P., Chojnacki, M., Załuska-Ogryzek, K., Kurzepa, J., & Łuszczki, J. J. (2025). Rasagiline Inhibits Human Melanoma Cell Viability and Interacts Synergistically with Mitoxantrone and Antagonistically with Cisplatin—In Vitro Isobolographic Studies. Cancers, 17(15), 2563. https://doi.org/10.3390/cancers17152563