Abstract
Cyclosporine A (CsA) is an immunosuppressant primarily used at a higher dosage in transplant medicine and autoimmune diseases with a higher success rate. At lower doses, CsA exhibits immunomodulatory properties. CsA has also been reported to inhibit breast cancer cell growth by downregulating the expression of pyruvate kinase. However, differential dose–response effects of CsA in cell growth, colonization, apoptosis, and autophagy remain largely unidentified in breast cancer cells. Herein, we showed the cell growth-inhibiting effects of CsA by preventing cell colonization and enhancing DNA damage and apoptotic index at a relatively lower concentration of 2 µM in MCF-7 breast cancer cells. However, at a higher concentration of 20 µM, CsA leads to differential expression of autophagy-related genes ATG1, ATG8, and ATG9 and apoptosis-associated markers, such as Bcl-2, Bcl-XL, Bad, and Bax, indicating a dose–response effect on differential cell death mechanisms in MCF-7 cells. This was confirmed in the protein–protein interaction network of COX-2 (PTGS2), a prime target of CsA, which had close interactions with Bcl-2, p53, EGFR, and STAT3. Furthermore, we investigated the combined effect of CsA with SHP2/PI3K-AKT inhibitors showing significant MCF-7 cell growth reduction, suggesting its potential to use as an adjuvant during breast cancer therapy.
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References
Borel JF, Feurer C, Gubler HU, Stähelin H (1976) Biological effects of cyclosporin A: a new antilymphocytic agent. Agents Actions 6:468–475. https://doi.org/10.1007/BF01973261
Borel JF, Feurer C, Magnée C, Stähelin H (1977) Effects of the new anti-lymphocytic peptide cyclosporin A in animals. Immunology 32:1017–1025
Cevik O, Turut FA, Acidereli H, Yildirim S (2019) Cyclosporine-A induces apoptosis in human prostate cancer cells PC3 and DU145 via downregulation of COX-2 and upregulation of TGFβ. Turk J Biochem 44:47–54. https://doi.org/10.1515/tjb-2017-0355
Chen Y-NP, LaMarche MJ, Chan HM et al (2016) Allosteric inhibition of SHP2 phosphatase inhibits cancers driven by receptor tyrosine kinases. Nature 535:148–152. https://doi.org/10.1038/nature18621
Ciechomska IA, Gabrusiewicz K, Szczepankiewicz AA, Kaminska B (2013) Endoplasmic reticulum stress triggers autophagy in malignant glioma cells undergoing cyclosporine A-induced cell death. Oncogene 32:1518–1529. https://doi.org/10.1038/onc.2012.174
Dhyani P, Quispe C, Sharma E et al (2022) Anticancer potential of alkaloids: a key emphasis to colchicine, vinblastine, vincristine, vindesine, vinorelbine and vincamine. Cancer Cell Int 22:206. https://doi.org/10.1186/s12935-022-02624-9
Durnian JM, Stewart RMK, Tatham R et al (2007) Cyclosporin-A associated malignancy. Clin Ophthalmol 1:421–430
Ershad A, Taziki S, Ebrahimian M, Abadi SSD (2023) Acute cyclosporine overdose: a systematic review. Medicina Clínica Práctica 6:100358. https://doi.org/10.1016/j.mcpsp.2022.100358
Etti I, Rasedee A, Mohd Hashim N et al (2017) Artonin E induces p53-independent G1 cell cycle arrest and apoptosis through ROS-mediated mitochondrial pathway and livin suppression in MCF-7 cells. DDDT 11:865–879. https://doi.org/10.2147/DDDT.S124324
Flores C, Fouquet G, Moura IC et al (2019) Lessons to learn from low-dose cyclosporin-A: a new approach for unexpected clinical applications. Front Immunol 10:588. https://doi.org/10.3389/fimmu.2019.00588
Forde K, Resta N, Ranieri C et al (2021) Clinical experience with the AKT1 inhibitor miransertib in two children with PIK3CA-related overgrowth syndrome. Orphanet J Rare Dis 16:109. https://doi.org/10.1186/s13023-021-01745-0
Frasor J, Weaver AE, Pradhan M, Mehta K (2008) Synergistic up-regulation of prostaglandin E synthase expression in breast cancer cells by 17β-estradiol and proinflammatory cytokines. Endocrinology 149:6272–6279. https://doi.org/10.1210/en.2008-0352
Gandalovičová A, Rosel D, Fernandes M et al (2017) Migrastatics—anti-metastatic and anti-invasion drugs: promises and challenges. Trends Cancer 3:391–406. https://doi.org/10.1016/j.trecan.2017.04.008
George VC, Kumar DRN, Suresh PK et al (2013) Comparative studies to evaluate relative in vitro potency of luteolin in inducing cell cycle arrest and apoptosis in HaCaT and A375 cells. Asian Pac J Cancer Prev 14:631–637. https://doi.org/10.7314/APJCP.2013.14.2.631
George VC, Dellaire G, Rupasinghe HPV (2017) Plant flavonoids in cancer chemoprevention: role in genome stability. J Nutr Biochem 45:1–14. https://doi.org/10.1016/j.jnutbio.2016.11.007
Groenendyk J, Paskevicius T, Urra H et al (2018) Cyclosporine A binding to COX-2 reveals a novel signaling pathway that activates the IRE1α unfolded protein response sensor. Sci Rep 8:16678. https://doi.org/10.1038/s41598-018-34891-w
Guo Z, Jiang J-H, Zhang J et al (2015) COX-2 promotes migration and invasion by the side population of cancer stem cell-like hepatocellular carcinoma cells. Medicine 94:e1806. https://doi.org/10.1097/MD.0000000000001806
Harris RE (2014) Cyclooxygenase-2 and the inflammogenesis of breast cancer. WJCO 5:677. https://doi.org/10.5306/wjco.v5.i4.677
Hojo M, Morimoto T, Maluccio M et al (1999) Cyclosporine induces cancer progression by a cell-autonomous mechanism. Nature 397:530–534. https://doi.org/10.1038/17401
Ioele G, Chieffallo M, Occhiuzzi MA et al (2022) Anticancer drugs: recent strategies to improve stability profile. Pharmacokinet Pharmacodyn Prop Mol 27:5436. https://doi.org/10.3390/molecules27175436
Jiang K, He B, Lai L et al (2012) Cyclosporine A inhibits breast cancer cell growth by downregulating the expression of pyruvate kinase subtype M2. Int J Mol Med 30:302–308. https://doi.org/10.3892/ijmm.2012.989
Jin B, Robertson KD (2013) DNA methyltransferases, DNA damage repair, and cancer. In: Karpf AR (ed) Epigenetic alterations in oncogenesis. Springer, New York, pp 3–29
Kawahara T, Kashiwagi E, Ide H et al (2015) Cyclosporine A and tacrolimus inhibit bladder cancer growth through down-regulation of NFATc1. Oncotarget 6:1582–1593. https://doi.org/10.18632/oncotarget.2750
Kim (2010) Cyclosporin A and sanglifehrin A enhance chemotherapeutic effect of cisplatin in C6 glioma cells. Oncol Rep. https://doi.org/10.3892/or_00000732
Kim HS, Choi S-I, Jeung E-B, Yoo Y-M (2014) Cyclosporine A induces apoptotic and autophagic cell death in rat pituitary GH3 cells. PLoS ONE 9:e108981. https://doi.org/10.1371/journal.pone.0108981
Kinner A, Wu W, Staudt C, Iliakis G (2008) H2AX in recognition and signaling of DNA double-strand breaks in the context of chromatin. Nucleic Acids Res 36:5678–5694. https://doi.org/10.1093/nar/gkn550
Kubben FJ, Peeters-Haesevoets A, Engels LG et al (1994) Proliferating cell nuclear antigen (PCNA): a new marker to study human colonic cell proliferation. Gut 35:530–535. https://doi.org/10.1136/gut.35.4.530
Lakshmikuttyamma A, Selvakumar P, Kanthan R et al (2005) Increased expression of calcineurin in human colorectal adenocarcinomas. J Cell Biochem 95:731–739. https://doi.org/10.1002/jcb.20437
Lally C, Healy E, Ryan MP (1999) Cyclosporine A-induced cell cycle arrest and cell death in renal epithelial cells. Kidney Int 56:1254–1257. https://doi.org/10.1046/j.1523-1755.1999.00696.x
Landewé RBM, van den Borne BEEM, Breedveld FC, Dijkmans BAC (1999) Does cyclosporin A cause cancer? Nat Med 5:714–714. https://doi.org/10.1038/10417
Laranjeira ABA, Hollingshead MG, Nguyen D et al (2023) DNA damage, demethylation and anticancer activity of DNA methyltransferase (DNMT) inhibitors. Sci Rep 13:5964. https://doi.org/10.1038/s41598-023-32509-4
Liu Z, Jiang L, Li Y et al (2019) Cyclosporine A sensitizes lung cancer cells to crizotinib through inhibition of the Ca2+/calcineurin/Erk pathway. EBioMedicine 42:326–339. https://doi.org/10.1016/j.ebiom.2019.03.019
Lomphithak T, Fadeel B (2023) Die hard: cell death mechanisms and their implications in nanotoxicology. Toxicol Sci 192:141–154. https://doi.org/10.1093/toxsci/kfad008
Lowe NJ, Wieder JM, Rosenbach A et al (1996) Long-term low-dose cyclosporine therapy for severe psoriasis: effects on renal function and structure. J Am Acad Dermatol 35:710–719. https://doi.org/10.1016/S0190-9622(96)90726-4
Masuo T, Okamura S, Zhang Y, Mori M (2009) Cyclosporine A inhibits colorectal cancer proliferation probably by regulating expression levels of c-Myc, p21WAF1/CIP1 and proliferating cell nuclear antigen. Cancer Lett 285:66–72. https://doi.org/10.1016/j.canlet.2009.05.001
Miach PJ (1986) Cyclosporin A in organ transplantation. Med J Aust 145:146–150. https://doi.org/10.5694/j.1326-5377.1986.tb113775.x
Mukherjee S, Mukherjee U (2009) A comprehensive review of immunosuppression used for liver transplantation. J Transplant 2009:1–20. https://doi.org/10.1155/2009/701464
Olivier T, Haslam A, Prasad V (2021) Anticancer drugs approved by the US Food and Drug Administration from 2009 to 2020 according to their mechanism of action. JAMA Netw Open 4:e2138793. https://doi.org/10.1001/jamanetworkopen.2021.38793
Papinski D, Schuschnig M, Reiter W et al (2014) Early steps in autophagy depend on direct phosphorylation of Atg9 by the Atg1 kinase. Mol Cell 53:471–483. https://doi.org/10.1016/j.molcel.2013.12.011
Patocka J, Nepovimova E, Kuca K, Wu W (2021) Cyclosporine A: chemistry and toxicity—a review. CMC 28:3925–3934. https://doi.org/10.2174/0929867327666201006153202
Popgeorgiev N, Jabbour L, Gillet G (2018) Subcellular localization and dynamics of the Bcl-2 family of proteins. Front Cell Dev Biol 6:13. https://doi.org/10.3389/fcell.2018.00013
Rehman AU, Rahman MU, Khan MT et al (2019) The landscape of protein tyrosine phosphatase (Shp2) and cancer. CPD 24:3767–3777. https://doi.org/10.2174/1381612824666181106100837
Scardoni G, Tosadori G, Faizan M et al (2015) Biological network analysis with CentiScaPe: centralities and experimental dataset integration. F1000Res 3:139. https://doi.org/10.12688/f1000research.4477.2
Scott RC, Juhász G, Neufeld TP (2007) Direct induction of autophagy by Atg1 inhibits cell growth and induces apoptotic cell death. Curr Biol 17:1–11. https://doi.org/10.1016/j.cub.2006.10.053
Siddiqui SS, Rahman S, Rupasinghe HPV, Vazhappilly CG (2020) Dietary flavonoids in p53—mediated immune dysfunctions linking to cancer prevention. Biomedicines 8:286. https://doi.org/10.3390/biomedicines8080286
Slobodkin MR, Elazar Z (2013) The Atg8 family: multifunctional ubiquitin-like key regulators of autophagy. Essays Biochem 55:51–64. https://doi.org/10.1042/bse0550051
Szklarczyk D, Franceschini A, Wyder S et al (2015) STRING v10: protein–protein interaction networks, integrated over the tree of life. Nucleic Acids Res 43:D447–D452. https://doi.org/10.1093/nar/gku1003
Tait SWG, Green DR (2013) Mitochondrial regulation of cell death. Cold Spring Harb Perspect Biol 5:a008706–a008706. https://doi.org/10.1101/cshperspect.a008706
Tan Y (2011) Effect of bisphenol A on the EGFR-STAT3 pathway in MCF-7 breast cancer cells. Mol Med Rep. https://doi.org/10.3892/mmr.2011.583
Tanaka Y, Tsujimura S (2002) Clinical implication of cyclosporin for rheumatoid arthritis. Nihon Rinsho 60:2345–2350
Thoms K-M, Kuschal C, Oetjen E et al (2011) Cyclosporin A, but not everolimus, inhibits DNA repair mediated by calcineurin: implications for tumorigenesis under immunosuppression: cyclosporin A inhibits DNA repair mediated by calcineurin. Exp Dermatol 20:232–236. https://doi.org/10.1111/j.1600-0625.2010.01213.x
Vazhappilly CG, Saleh E, Ramadan W et al (2019) Inhibition of SHP2 by new compounds induces differential effects on RAS/RAF/ERK and PI3K/AKT pathways in different cancer cell types. Investig New Drugs 37:252–261. https://doi.org/10.1007/s10637-018-0626-5
Vazhappilly CG, Hodeify R, Siddiqui SS et al (2020) Natural compound catechol induces DNA damage, apoptosis, and G1 cell cycle arrest in breast cancer cells. Phytother Res. https://doi.org/10.1002/ptr.6970
Walter DH, Haendeler J, Galle J et al (1998) Cyclosporin A inhibits apoptosis of human endothelial cells by preventing release of cytochrome C from mitochondria. Circulation 98:1153–1157. https://doi.org/10.1161/01.CIR.98.12.1153
Wang S, He M, Li L et al (2016) Cell-in-cell death is not restricted by caspase-3 deficiency in MCF-7 cells. J Breast Cancer 19:231–241. https://doi.org/10.4048/jbc.2016.19.3.231
Wee P, Wang Z (2017) Epidermal growth factor receptor cell proliferation signaling pathways. Cancers 9:52. https://doi.org/10.3390/cancers9050052
Werneck MBF, Hottz E, Bozza PT, Viola JPB (2012) Cyclosporin A inhibits colon cancer cell growth independently of the calcineurin pathway. Cell Cycle 11:3997–4008. https://doi.org/10.4161/cc.22222
Wu P-J, Hsin I-L, Hung W-L et al (2022) Combination treatment with cyclosporin A and arsenic trioxide induce synergistic cell death via non-apoptotic pathway in uterine cervical cancer cells. Chem Biol Interact 368:110177. https://doi.org/10.1016/j.cbi.2022.110177
Yuan Y, Fan Y, Gao Z et al (2020) SHP2 promotes proliferation of breast cancer cells through regulating Cyclin D1 stability via the PI3K/AKT/GSK3β signaling pathway. Cancer Biol Med 17:707–725. https://doi.org/10.20892/j.issn.2095-3941.2020.0056
Zhou X, Coad J, Ducatman B, Agazie YM (2008) SHP2 is up-regulated in breast cancer cells and in infiltrating ductal carcinoma of the breast, implying its involvement in breast oncogenesis. Histopathology 53:389–402. https://doi.org/10.1111/j.1365-2559.2008.03103.x
Zhuang X, Chung KP, Cui Y et al (2017) ATG9 regulates autophagosome progression from the endoplasmic reticulum in Arabidopsis. Proc Natl Acad Sci USA. https://doi.org/10.1073/pnas.1616299114
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Authors would like to thank the American University of Ras Al Khaimah for the support of this research.
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Siddiqui, S.S., Hodeify, R., Mathew, S. et al. Differential dose–response effect of cyclosporine A in regulating apoptosis and autophagy markers in MCF-7 cells. Inflammopharmacol 31, 2049–2060 (2023). https://doi.org/10.1007/s10787-023-01247-4
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DOI: https://doi.org/10.1007/s10787-023-01247-4