Low-Dose Salinomycin Targets Multidrug Resistance via P-Glycoprotein and NF-κB in Osteosarcoma Cells
DOI:
https://doi.org/10.26877/asset.v8i3.2447Keywords:
Multidrug resistance, NF-κB, osteosarcoma, P-Glycoprotein, salinomycinAbstract
Chemotherapy resistance in osteosarcoma remains a major clinical obstacle, largely driven by drug efflux mechanisms mediated by P-glycoprotein (ACBC1/ABCB1) and activation of NF-κB signaling (NFKB1). This study investigated the potential of Salinomycin to modulate these multidrug resistance–associated genes in U2OS osteosarcoma cells and explore its role in overcoming chemoresistance. A true experimental post-test-only control group design was used. U2OS cells were treated with varying concentrations of Salinomycin, doxorubicin, and their combination. Cell viability was assessed using MTT assay, while gene expression of ACBC1 and NFKB1 was quantified using qRT-PCR with SYBR Green chemistry. Relative expression levels were analyzed using the 2−ΔΔCt method normalized to GAPDH. Salinomycin demonstrated dose-dependent cytotoxic effects in the low micromolar range. At lower concentrations, it significantly reduced NFKB1 expression, while also showing a tendency to downregulate ACBC1. However, intermediate concentrations showed variable effects, including a transient increase in NFKB1 expression. The combination treatment with doxorubicin produced only modest and non-significant changes in both resistance-related genes. Overall, low-dose Salinomycin exhibited a more consistent suppressive effect on NF-κB signaling, suggesting a potential role in sensitizing osteosarcoma cells to chemotherapy. In contrast, higher doses primarily enhanced cytotoxicity without clearly improving suppression of resistance markers. This study highlights a novel dual-action, dose-dependent regulatory effect of Salinomycin at the transcriptomic level, targeting both drug efflux (ACBC1/ABCB1) and survival signaling (NF-κB/NFKB1). These findings provide new insight into its potential as an adjunct agent in overcoming multidrug resistance in osteosarcoma and warrant further validation at the protein and functional levels.References
[1] X. Zhao, Q. Wu, X. Gong, J. Liu, and Y. Ma, “Osteosarcoma: a review of current and future therapeutic approaches,” Biomed Eng Online, vol. 20, no. 1, p. 24, 2021.
[2] J. H. Alexander, O. T. Binitie, G. D. Letson, and D. M. Joyce, “Osteosarcoma: an evolving understanding of a complex disease,” JAAOS-Journal of the American Academy of Orthopaedic Surgeons, vol. 29, no. 20, pp. e993–e1004, 2021. 10.5435/JAAOS-D-20-00838
[3] M. Li and W. Ma, “miR-26a reverses multidrug resistance in osteosarcoma by targeting MCL1,” Front Cell Dev Biol, vol. 9, p. 645381, 2021. https://doi.org/10.3389/fcell.2021.645381
[4] L. Marchandet, M. Lallier, C. Charrier, M. Baud’huin, B. Ory, and F. Lamoureux, “Mechanisms of resistance to conventional therapies for osteosarcoma,” Cancers (Basel), vol. 13, no. 4, p. 683, 2021. https://doi.org/10.3390/cancers13040683
[5] T. Liu et al., “Targeting ABCB1 (MDR1) in multi-drug resistant osteosarcoma cells using the CRISPR-Cas9 system to reverse drug resistance,” Oncotarget, vol. 7, no. 50, p. 83502, 2016. 10.18632/oncotarget.13148
[6] F. Klepsch, P. Vasanthanathan, and G. F. Ecker, “Ligand and structure-based classification models for prediction of P-glycoprotein inhibitors,” J Chem Inf Model, vol. 54, no. 1, pp. 218–229, 2014.
[7] P. B. Gupta et al., “Identification of selective inhibitors of cancer stem cells by high-throughput screening,” Cell, vol. 138, no. 4, pp. 645–659, 2009. 10.1016/j.cell.2009.06.034
[8] C. Naujokat and R. Steinhart, “Salinomycin as a drug for targeting human cancer stem cells,” Biomed Res Int, vol. 2012, no. 1, p. 950658, 2012. https://doi.org/10.1155/2012/950658
[9] K.-Y. Kim et al., “Salinomycin-induced apoptosis of human prostate cancer cells due to accumulated reactive oxygen species and mitochondrial membrane depolarization,” Biochem Biophys Res Commun, vol. 413, no. 1, pp. 80–86, 2011. https://doi.org/10.1016/j.bbrc.2011.08.054
[10] J. Kim et al., “Salinomycin sensitizes cancer cells to the effects of doxorubicin and etoposide treatment by increasing DNA damage and reducing p21 protein,” Br J Pharmacol, vol. 162, no. 3, pp. 773–784, 2011. https://doi.org/10.1111/j.1476-5381.2010.01089.x
[11] B. Verdoodt, M. Vogt, I. Schmitz, S.-T. Liffers, A. Tannapfel, and A. Mirmohammadsadegh, “Salinomycin induces autophagy in colon and breast cancer cells with concomitant generation of reactive oxygen species,” 2012. https://doi.org/10.1371/journal.pone.0044132
[12] K.-Y. Kim et al., “Salinomycin induces reactive oxygen species and apoptosis in aggressive breast cancer cells as mediated with regulation of autophagy,” Anticancer Res, vol. 37, no. 4, pp. 1747–1758, 2017.
[13] R. Riccioni et al., “The cancer stem cell selective inhibitor Salinomycin is a p-glycoprotein inhibitor,” Blood Cells Mol Dis, vol. 45, no. 1, pp. 86–92, 2010. https://doi.org/10.1016/j.bcmd.2010.03.008
[14] D. Fuchs, A. Heinold, G. Opelz, V. Daniel, and C. Naujokat, “Salinomycin induces apoptosis and overcomes apoptosis resistance in human cancer cells,” Biochem Biophys Res Commun, vol. 390, no. 3, pp. 743–749, 2009. https://doi.org/10.1016/j.bbrc.2009.10.042
[15] J. Dewangan, S. Srivastava, and S. K. Rath, “Salinomycin: A new paradigm in cancer therapy,” Tumor Biology, vol. 39, no. 3, p. 1010428317695035, 2017. https://doi.org/10.1177/1010428317695035
[16] K. J. Livak and T. D. Schmittgen, “Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method,” methods, vol. 25, no. 4, pp. 402–408, 2001. https://doi.org/10.1006/meth.2001.1262
[17] A. Seelig, “P-glycoprotein: one mechanism, many tasks and the consequences for pharmacotherapy of cancers,” Front Oncol, vol. 10, p. 576559, 2020. https://doi.org/10.3389/fonc.2020.576559
[18] S. Mirzaei et al., “Advances in understanding the role of P-gp in doxorubicin resistance: Molecular pathways, therapeutic strategies, and prospects,” Drug Discov Today, vol. 27, no. 2, pp. 436–455, 2022. 10.1016/j.drudis.2021.09.020
[19] L. Yang, D. Li, P. Tang, and Y. Zuo, “Curcumin increases the sensitivity of K562/DOX cells to doxorubicin by targeting S100 calcium-binding protein A8 and P-glycoprotein,” Oncol Lett, vol. 19, no. 1, pp. 83–92, 2020. https://doi.org/10.3892/ol.2019.11083
[20] Cheng P, Liao HY, and Zhang HH, “Role of NF-κB Signaling Pathway in Osteosarcoma,” Lupine Online Journal of Medical Sciences, 2022.
[21] M. Tyagi and B. S. Patro, “Salinomycin reduces growth, proliferation and metastasis of cisplatin resistant breast cancer cells via NF-kB deregulation,” Toxicology in Vitro, vol. 60, pp. 125–133, 2019. 10.1016/j.tiv.2019.05.004
[22] Y. Lu, F. Li, T. Xu, and J. Sun, “Tetrandrine prevents multidrug resistance in the osteosarcoma cell line, U-2OS, by preventing Pgp overexpression through the inhibition of NF-κB signaling,” Int J Mol Med, vol. 39, no. 4, pp. 993–1000, 2017. [20] Cheng P, Liao HY, and Zhang HH, “Role of NF-κB Signaling Pathway in Osteosarcoma,” Lupine Online Journal of Medical Sciences, 2022. https://doi.org/10.3892/ijmm.2017.2895
[23] N. B. Antoun and A.-M. Chioni, “Dysregulated Signalling Pathways Driving Anticancer Drug Resistance,” International Journal of Molecular Sciences, vol. 24, no. 15, p. 12222, Jul. 2023, https://doi.org/10.3390/ijms241512222
[24] S. U. Khan, K. Fatima, S. Aisha, and F. Malik, “Unveiling the mechanisms and challenges of cancer drug resistance,” Cell Communication and Signaling, vol. 22, art. no. 109, Feb. 2024, doi: 10.1186/s12964-023-01302-1.
[25] J. Song et al., “Therapeutic effects of tetrandrine in inflammatory diseases: a comprehensive review,” Inflammopharmacology, vol. 32, pp. 1743–1757, Apr. 2024, doi: 10.1007/s10787-024-01452-9.
Downloads
Published
Issue
Section
License
Copyright (c) 2026 Advance Sustainable Science Engineering and Technology
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
