Biomolecules & Therapeutics

The Korean Society of Applied Pharmacology (한국응용약물학회)
권호 표지
DOI QR코드

DOI QR Code

β-Catenin and AMPK/AKT/FOXO Signaling Mediate Doxorubicin-Induced Senescence and Lipid Accumulation in C2C12 Myoblasts

  • Chawon Yun (Department of Pharmacy, College of Pharmacy, Research Institute for Drug Development, Pusan National University) ;
  • Sou Hyun Kim (Department of Pharmacy, College of Pharmacy, Research Institute for Drug Development, Pusan National University) ;
  • Doyoung Kwon (College of Pharmacy, Jeju Research Institute of Pharmaceutical Sciences, Jeju National University) ;
  • RanJu Woo (Department of Pharmacy, College of Pharmacy, Research Institute for Drug Development, Pusan National University) ;
  • Ki Wung Chung (Department of Pharmacy, College of Pharmacy, Research Institute for Drug Development, Pusan National University) ;
  • Jaewon Lee (Department of Pharmacy, College of Pharmacy, Research Institute for Drug Development, Pusan National University) ;
  • Yun-Hee Lee (College of Pharmacy, Research Institute of Pharmaceutical Sciences, Seoul National University) ;
  • Young-Suk Jung (Department of Pharmacy, College of Pharmacy, Research Institute for Drug Development, Pusan National University)
  • Received : 2025.10.27
  • Accepted : 2025.12.09
  • Published : 2026.01.01
Download PDF
⟨ Previous Next ⟩

Abstract

Skeletal muscle atrophy is a major complication associated with aging, chronic disease, and chemotherapy. Doxorubicin (Dox), a widely used anticancer agent, accelerates muscle wasting; however, the underlying cellular mechanisms remain poorly understood. In this study, we examined the effects of Dox on myogenic differentiation, senescence, and lipid metabolism using C2C12 myoblasts. Dox exposure impaired myotube formation without causing overt cytotoxicity. Mechanistically, Dox disrupted myogenic differentiation by inhibiting protein kinase B/mammalian target of rapamycin (AKT/mTOR) signaling, thereby de-repressing forkhead box O1/3 (FOXO1/3) and upregulating the muscle-specific ubiquitin ligases muscle atrophy F-box (MAFbx) and muscle RING finger 1 (MuRF1), which promote proteolysis. Dox also decreased glycogen synthase kinase 3β (GSK3β) phosphorylation while paradoxically increasing total and phosphorylated β-catenin, indicating dysregulated Wnt/β-catenin signaling. These alterations were accompanied by a senescence-like phenotype, characterized by elevated senescence-associated β-galactosidase (SA-β-gal) activity, increased phosphorylated histone variant γH2AX, and activation of the p53-p21 axis. Notably, cellular senescence coincided with excessive lipid accumulation in myotubes. Dox reduced phosphorylation of adenosine monophosphate-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) while enhancing expression of key lipogenic regulators, thereby creating a metabolic environment favoring lipid storage. Collectively, these findings demonstrate that Dox not only suppresses myogenic differentiation but also induces premature senescence and metabolic reprogramming toward lipid accumulation. Targeting these pathways through AMPK activation, FOXO inhibition, or senolytic interventions may offer therapeutic strategies to preserve skeletal muscle integrity in patients undergoing chemotherapy.

Keywords

Acknowledgement

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Education (2019R1A6A1A10072987). This work was also supported by the Bio & Medical Technology Development Program of the National Research Foundation (NRF), funded by the Korean government (MSIT) (RS-2025-16063709). It was also supported by a grant from the National Research Foundation of Korea (NRF), funded by the Ministry of Science and ICT (RS-2025-00558179).

References

  1. Ajoolabady, A., Pratico, D., Bahijri, S., Tuomilehto, J., Uversky, V. N. and Ren, J. (2025) Hallmarks of cellular senescence: biology, mechanisms, regulations. Exp. Mol. Med. 57, 1482-1491. https://doi.org/10.1038/s12276-025-01480-7
  2. Beloribi-Djefaflia, S., Vasseur, S. and Guillaumond, F. (2016) Lipid metabolic reprogramming in cancer cells. Oncogenesis 5, e189. https://doi.org/10.1038/oncsis.2015.49
  3. Bielak-Zmijewska, A., Wnuk, M., Przybylska, D., Grabowska, W., Lewinska, A., Alster, O., Korwek, Z., Cmoch, A., Myszka, A., Pikula, S., Mosieniak, G. and Sikora, E. (2014) A comparison of replicative senescence and doxorubicin-induced premature senescence of vascular smooth muscle cells isolated from human aorta. Biogerontology 15, 47-64. https://doi.org/10.1007/s10522-013-9477-9
  4. Byun, M. R., Kim, S. H., Woo, R., Noh, S. J., Joo, S. H., Jung, Y. S. and Choi, J. S. (2025) Quinine inhibits myogenic differentiation by disrupting AKT signaling pathway. Toxicol. Res. 41, 81-90. https://doi.org/10.1007/s43188-024-00273-w
  5. Chang, Z. S., Wang, L., Zhou, J. X., Li, M. X., Yang, M. Y., Luo, B., Liu, J. J., Sun, X. Y. and Xia, J. B. (2025) FoxO3 activation alleviates doxorubicin-induced cardiomyopathy by enhancing autophagic flux and suppressing mTOR/ROS signalling. J. Cell. Mol. Med. 29, e70775. https://doi.org/10.1111/jcmm.v29.15
  6. Chen, K., Gao, P., Li, Z., Dai, A., Yang, M., Chen, S., Su, J., Deng, Z. and Li, L. (2022) Forkhead box O signaling pathway in skeletal muscle atrophy. Am. J. Pathol. 192, 1648-1657. https://doi.org/10.1016/j.ajpath.2022.09.003
  7. Chen, P., Jia, F., Wang, M. and Yang, S. (2025) Analysis of the mechanism of skeletal muscle atrophy from the pathway of decreased protein synthesis. Front. Physiol. 16, 1533394.
  8. Coletti, D. (2018) Chemotherapy-induced muscle wasting: an update. Eur. J. Transl. Myol. 28, 7587. https://doi.org/10.4081/ejtm.2018.7587
  9. Dungan, C. M., Wells, J. M. and Murach, K. A. (2023) The life and times of cellular senescence in skeletal muscle: friend or foe for homeostasis and adaptation? Am. J. Physiol. Cell Physiol. 325, C324-C331. https://doi.org/10.1152/ajpcell.00553.2022
  10. Faber, J. G., van Herwijnen, M., Hauser, D., Daemen, S., Caiment, F. and van den Beucken, T. (2025) FOXO3 mediated gene expression modulates doxorubicin sensitivity in human cardiomyocytes. Toxicology 518, 154267. https://doi.org/10.1016/j.tox.2025.154267
  11. Garcia, D. and Shaw, R. J. (2017) AMPK: mechanisms of cellular energy sensing and restoration of metabolic balance. Mol. Cell 66, 789-800. https://doi.org/10.1016/j.molcel.2017.05.032
  12. Gilliam, L. A. and St Clair, D. K. (2011) Chemotherapy-induced weakness and fatigue in skeletal muscle: the role of oxidative stress. Antioxid. Redox Signal. 15, 2543-2563. https://doi.org/10.1089/ars.2011.3965
  13. Goyal, A., Afzal, M., Khan, N. H., Goyal, K., Srinivasamurthy, S. K., Gupta, G., Benod Kumar, K., Ali, H., Rana, M., Wong, L. S., Kumarasamy, V. and Subramaniyan, V. (2025) Targeting p53-p21 signaling to enhance mesenchymal stem cell regenerative potential. Regen. Ther. 29, 352-363. https://doi.org/10.1016/j.reth.2025.03.007
  14. Hamrick, M. W., McGee-Lawrence, M. E. and Frechette, D. M. (2016) Fatty infiltration of skeletal muscle: mechanisms and comparisons with bone marrow adiposity. Front. Endocrinol. 7, 69. https://doi.org/10.3389/fendo.2016.00069
  15. Hamsanathan, S. and Gurkar, A. U. (2022) Lipids as regulators of cellular senescence. Front. Physiol. 13, 796850.
  16. Hiensch, A. E., Bolam, K. A., Mijwel, S., Jeneson, J. A. L., Huitema, A. D. R., Kranenburg, O., van der Wall, E., Rundqvist, H., Wengstrom, Y. and May, A. M. (2020) Doxorubicin-induced skeletal muscle atrophy: Elucidating the underlying molecular pathways. Acta Physiol. (Oxf.) 229, e13400. https://doi.org/10.1111/apha.v229.2
  17. Huang, S. C., Wu, J. F., Saovieng, S., Chien, W. H., Hsu, M. F., Li, X. F., Lee, S. D., Huang, C. Y., Huang, C. Y. and Kuo, C. H. (2017) Doxorubicin inhibits muscle inflammation after eccentric exercise. J. Cachexia Sarcopenia Muscle 8, 277-284. https://doi.org/10.1002/jcsm.v8.2
  18. Jackman, R. W. and Kandarian, S. C. (2004) The molecular basis of skeletal muscle atrophy. Am. J. Physiol. Cell Physiol. 287, C834-C843. https://doi.org/10.1152/ajpcell.00579.2003
  19. Jaiswal, N., Gavin, M., Loro, E., Sostre-Colon, J., Roberson, P. A., Uehara, K., Rivera-Fuentes, N., Neinast, M., Arany, Z., Kimball, S. R., Khurana, T. S. and Titchenell, P. M. (2022) AKT controls protein synthesis and oxidative metabolism via combined mTORC1 and FOXO1 signalling to govern muscle physiology. J. Cachexia Sarcopenia Muscle 13, 495-514. https://doi.org/10.1002/jcsm.v13.1
  20. Jensen-Urstad, A. P. and Semenkovich, C. F. (2012) Fatty acid synthase and liver triglyceride metabolism: housekeeper or messenger? Biochim. Biophys. Acta 1821, 747-753. https://doi.org/10.1016/j.bbalip.2011.09.017
  21. Jones, A. E., Price, F. D., Le Grand, F., Soleimani, V. D., Dick, S. A., Megeney, L. A. and Rudnicki, M. A. (2015) Wnt/beta-catenin controls follistatin signalling to regulate satellite cell myogenic potential. Skelet. Muscle 5, 14.
  22. Kajabadi, N., Low, M., Jacques, E., Lad, H., Tung, L. W., Babaeijandaghi, F., Gamu, D., Zelada, D., Wong, C. K., Chang, C., Yi, L., Wosczyna, M. N., Rando, T. A., Henríquez, J. P., Gibson, W. T., Gilbert, P. M. and Rossi, F. M. V. (2023) Activation of β-catenin in mesenchymal progenitors leads to muscle mass loss. Dev. Cell 58, 489-505.e7. https://doi.org/10.1016/j.devcel.2023.02.009
  23. Kumari, R. and Jat, P. (2021) Mechanisms of cellular senescence: cell cycle arrest and senescence associated secretory phenotype. Front. Cell Dev. Biol. 9, 645593.
  24. Li, C. W., Yu, K., Shyh-Chang, N., Jiang, Z., Liu, T., Ma, S., Luo, L., Guang, L., Liang, K., Ma, W., Miao, H., Cao, W., Liu, R., Jiang, L. J., Yu, S. L., Li, C., Liu, H. J., Xu, L. Y., Liu, R. J., Zhang, X. Y. and Liu, G. S. (2022) Pathogenesis of sarcopenia and the relationship with fat mass: descriptive review. J. Cachexia Sarcopenia Muscle 13, 781-794. https://doi.org/10.1002/jcsm.v13.2
  25. Li, M., Ding, L., Cao, L., Zhang, Z., Li, X., Li, Z., Xia, Q., Yin, K., Song, S., Wang, Z., Du, H., Zhao, D., Li, X. and Wang, Z. (2025) Natural products targeting AMPK signaling pathway therapy, diabetes mellitus and its complications. Front. Pharmacol. 16, 1534634.
  26. Li, W., Gan, C., Yu, S., Xu, J., Tang, L. and Cheng, H. (2024a) Wnt3a/GSK3β/β-catenin signalling modulates doxorubicin-associated memory deficits in breast cancer. Mol. Neurobiol. 61, 5441-5458.
  27. Li, Y., Yan, J. and Yang, P. (2024b) The mechanism and therapeutic strategies in doxorubicin-induced cardiotoxicity: role of programmed cell death. Cell Stress Chaperones 29, 666-680.
  28. Li, Y., Zhang, Y., Li, R., Chen, W., Howell, M., Zhang, R. and Chen, G. (2012) The hepatic Raldh1 expression is elevated in Zucker fatty rats and its over-expression introduced the retinal-induced Srebp-1c expression in INS-1 cells. PLoS One 7, e45210. https://doi.org/10.1371/journal.pone.0045210
  29. Liu, B., Peng, Z., Zhang, H., Zhang, N., Liu, Z., Xia, Z., Huang, S., Luo, P. and Cheng, Q. (2025) Regulation of cellular senescence in tumor progression and therapeutic targeting: mechanisms and pathways. Mol. Cancer 24, 106.
  30. Liu, J., Xiao, Q., Xiao, J., Niu, C., Li, Y., Zhang, X., Zhou, Z., Shu, G. and Yin, G. (2022) Wnt/β-catenin signalling: function, biological mechanisms, and therapeutic opportunities. Signal. Transduct. Target. Ther. 7, 3.
  31. Matsui, H., Yokoyama, T., Sekiguchi, K., Iijima, D., Sunaga, H., Maniwa, M., Ueno, M., Iso, T., Arai, M. and Kurabayashi, M. (2012) Stearoyl-CoA desaturase-1 (SCD1) augments saturated fatty acid induced lipid accumulation and inhibits apoptosis in cardiac myocytes. PLoS One 7, e33283. https://doi.org/10.1371/journal.pone.0033283
  32. Nissinen, T. A., Degerman, J., Räsänen, M., Poikonen, A. R., Koskinen, S., Mervaala, E., Pasternack, A., Ritvos, O., Kivelä, R. and Hulmi, J. J. (2016) Systemic blockade of ACVR2B ligands prevents chemotherapy-induced muscle wasting by restoring muscle protein synthesis without affecting oxidative capacity or atrogenes. Sci. Rep. 6, 32695. https://doi.org/10.1038/srep32695
  33. Ohkawa, Y., Marfella, C. G. and Imbalzano, A. N. (2006) Skeletal muscle specification by myogenin and Mef2D via the SWI/SNF ATPase Brg1. EMBO J. 25, 490-501. https://doi.org/10.1038/sj.emboj.7600943
  34. Pigg, Q. W., Harris, D. R., Inoue, D. S. and Janini Gomes, M. (2025) Mitigating doxorubicin-induced skeletal muscle toxicity: a review of oxidative stress mechanisms and the therapeutic role of exercise. Antioxidants (Basel) 14, 45. https://doi.org/10.3390/antiox14070870
  35. Pundlik, S. S., Barik, A., Venkateshvaran, A., Sahoo, S. S., Jaysingh, M. A., Math, R. G. H., Lal, H., Hashmi, M. A. and Ramanathan, A. (2024) Senescent cells inhibit mouse myoblast differentiation via the SASP-lipid 15d-PGJ(2) mediated modification and control of HRas. Elife 13, RP95229.
  36. Saito, Y. and Chikenji, T. S. (2021) Diverse roles of cellular senescence in skeletal muscle inflammation, regeneration, and therapeutics. Front. Pharmacol. 12, 739510.
  37. Sartori, R., Romanello, V. and Sandri, M. (2021) Mechanisms of muscle atrophy and hypertrophy: implications in health and disease. Nat. Commun. 12, 330. https://doi.org/10.1038/s41467-020-20123-1
  38. Segalés, J., Perdiguero, E., Serrano, A. L., Sousa-Victor, P., Ortet, L., Jardí, M., Budanov, A. V., Garcia-Prat, L., Sandri, M., Thomson, D. M., Karin, M., Hee Lee, J. and Muñoz-Cánoves, P. (2020) Sestrin prevents atrophy of disused and aging muscles by integrating anabolic and catabolic signals. Nat. Commun. 11, 189.
  39. Seo, J., Kwon, D., Kim, S. H., Byun, M. R., Lee, Y. H. and Jung, Y. S. (2024) Role of autophagy in betaine-promoted hepatoprotection against non-alcoholic fatty liver disease in mice. Curr. Res. Food Sci. 8, 100663.
  40. Stitt, T. N., Drujan, D., Clarke, B. A., Panaro, F., Timofeyva, Y., Kline, W. O., Gonzalez, M., Yancopoulos, G. D. and Glass, D. J. (2004) The IGF-1/PI3K/Akt pathway prevents expression of muscle atrophy-induced ubiquitin ligases by inhibiting FOXO transcription factors. Mol. Cell 14, 395-403. https://doi.org/10.1016/S1097-2765(04)00211-4
  41. Van Asbroeck, B., Krüger, D. N., Van den Bogaert, S., Dombrecht, D., Bosman, M., Van Craenenbroeck, E. M., Guns, P. J. and van Breda, E. (2025) Distinct impact of doxorubicin on skeletal muscle and fat metabolism in mice: without dexrazoxane effect. Int. J. Mol. Sci. 26, 789.
  42. von Maltzahn, J., Chang, N. C., Bentzinger, C. F. and Rudnicki, M. A. (2012) Wnt signaling in myogenesis. Trends Cell Biol. 22, 602-609. https://doi.org/10.1016/j.tcb.2012.07.008
  43. Wang, D., Liu, Y., Zhang, R., Zhang, F., Sui, W., Chen, L., Zheng, R., Chen, X., Wen, F., Ouyang, H.-W. and Ji, J. (2016) Apoptotic transition of senescent cells accompanied with mitochondrial hyperfunction. Oncotarget 7, 28286-28300. https://doi.org/10.18632/oncotarget.v7i19
  44. Wang, Q., Liu, S., Zhai, A., Zhang, B. and Tian, G. (2018) AMPKmediated regulation of lipid metabolism by phosphorylation. Biol. Pharm. Bull. 41, 985-993. https://doi.org/10.1248/bpb.b17-00724
  45. Wiedmer, P., Jung, T., Castro, J. P., Pomatto, L. C. D., Sun, P. Y., Davies, K. J. A. and Grune, T. (2021) Sarcopenia - Molecular mechanisms and open questions. Ageing Res. Rev. 65, 101200.
  46. Xue, C., Chu, Q., Shi, Q., Zeng, Y., Lu, J. and Li, L. (2025) Wnt signaling pathways in biology and disease: mechanisms and therapeutic advances. Signal Transduct. Target. Ther. 10, 106.
  47. Yun, C., Kim, S. H., Kwon, D., Byun, M. R., Chung, K. W., Lee, J. and Jung, Y. S. (2024) Doxorubicin attenuates free fatty acid-induced lipid accumulation via stimulation of p53 in HepG2 cells. Biomol. Ther. (Seoul) 32, 94-103. https://doi.org/10.4062/biomolther.2023.200
  48. Zhang, F., Guo, J., Yu, S., Zheng, Y., Duan, M., Zhao, L., Wang, Y., Yang, Z. and Jiang, X. (2024) Cellular senescence and metabolic reprogramming: unraveling the intricate crosstalk in the immunosuppressive tumor microenvironment. Cancer Commun. (Lond.) 44, 929-966. https://doi.org/10.1002/cac2.12591
  49. Zhao, H., Ming, T., Tang, S., Ren, S., Yang, H., Liu, M., Tao, Q. and Xu, H. (2022) Wnt signaling in colorectal cancer: pathogenic role and therapeutic target. Mol. Cancer 21, 144.
  50. Zhou, X., Li, S., Wang, L., Wang, J., Zhang, P. and Chen, X. (2025) The emerging role of exercise preconditioning in preventing skeletal muscle atrophy. Front. Physiol. 16, 1559594.