Bavachalcone targets transferrin receptor and sensitizes gemcitabine to affect bladder cancer progression

Abstract

Gemcitabine resistance drives bladder cancer recurrence and progression. Using high-throughput drug screening in bladder cancer cells, we identified Bavachalcone (Bava) as a potent gemcitabine sensitizer. Mechanistically, Bava simultaneously targets transferrin receptor (TFRC) and epidermal growth factor receptor (EGFR). It competes with transferrin (Tf) for TFRC binding, reducing cellular iron influx, and inhibits EGFR-mediated phosphorylation of TFRC at tyrosine 20 (Y20). These actions disrupt mitochondria iron utilization and impairs respiration. The combination of Bava and gemcitabine synergistically inhibits the repair of gemcitabine-induced DNA damage, while suppressing the iron-dependent ATR-CHEK1-E2F1 pathway and downregulating RRM1 expression. Patient-derived xenograft models confirmed the superior antitumor efficacy of the Bava-gemcitabine co-treatment compared to monotherapies. Clinically, elevated TFRC and RRM1 expression correlates with poor prognosis, supporting their utility as biomarkers of bladder cancer. Our study identified Bava as the first small-molecule TFRC inhibitor that overcomes gemcitabine resistance through iron modulation, providing both mechanistic insights and a promising therapeutic strategy for bladder cancer.

Keywords

Bavachalcone / bladder cancer / DNA damage repair / gemcitabine resistance / TFRC and RRM1

References

[1] Zhang, Zi-hao, Yue Wang, Ya Zhang, Sheng-Feng Zheng, Tao Feng, Xi Tian, Mierxiati Abudurexiti, et al. 2023. “The Function and Mechanisms of Action of Circular RNAs in Urologic Cancer.” Molecular Cancer 22: 61. https://doi.org/10.1186/s12943-023-01766-2

[2] Galsky, Matthew D., Johannes Alfred Witjes, Jürgen E. Gschwend, Matthew I. Milowsky, Michael Schenker, Begoña P. Valderrama, Yoshihiko Tomita, et al. 2025. “Adjuvant Nivolumab in High-Risk Muscle-Invasive Urothelial Carcinoma: Expanded Efficacy From CheckMate 274.” Journal of Clinical Oncology 43: 15-21. https://doi.org/10.1200/JCO.24.00340

[3] Catto, J. W. F., B. Tran, M. Rouprêt, J. E. Gschwend, Y. Loriot, H. Nishiyama, J. P. Redorta, et al. 2024. “Erdafitinib in BCG-treated High-Risk Non-Muscle-Invasive Bladder Cancer.” Annals of Oncology 35: 98-106. https://doi.org/10.1016/j.annonc.2023.09.3116

[4] Chou, Roger, Shelley Selph, David I. Buckley, Rongwei Fu, Jessica C. Griffin, Sara Grusing, and John L. Gore. 2017. “Intravesical Therapy for the Treatment of Nonmuscle Invasive Bladder Cancer: A Systematic Review and Meta-Analysis.” Journal of Urology 197: 1189-1199. https://doi.org/10.1016/j.juro.2016.12.090

[5] Chen, Feiran, Jun Du, Xusheng Chen, Qing Yang, Lei Diao, and Xin Yao. 2024. “Efficacy and Safety of Short-Term Intravesical Gemcitabine vs. Long-Term Intravesical Epirubicin in the Treatment of Moderate to High-Risk Non-Muscle-Invasive Bladder Cancer: A Pilot Study.” Holistic Integrative Oncology 3: 8. https://doi.org/10.1007/s44178-024-00076-x

[6] Zhao, Jiaqi, Byunghyun Ahn, and Hening Lin. 2024. “Loss of Diphthamide Increases DNA Replication Stress in Mammalian Cells by Modulating the Translation of RRM1.” ACS Central Science 10: 1835-1847. https://doi.org/10.1021/acscentsci.4c00967

[7] Qin, Li, Zizheng Dong, Caoqinglong Huang, Hao Liu, Jenny Beebe, Boopathi Subramaniyan, Yangyang Hao, et al. 2023. “Reversible Promoter Demethylation of PDGFD Confers Gemcitabine Resistance Through STAT3 Activation and RRM1 Upregulation.” Cancer Letters 567: 216266. https://doi.org/10.1016/j.canlet.2023.216266

[8] Caverzan, Matías D., and Luis E. Ibarra. 2024. “Advancing Glioblastoma Treatment Through Iron Metabolism: A Focus on TfR1 and Ferroptosis Innovations.” International Journal of Biological Macromolecules 278: 134777. https://doi.org/10.1016/j.ijbiomac.2024.134777

[9] Packer, Milton. 2024. “Iron Homeostasis, Recycling and Vulnerability in the Stressed Kidney: A Neglected Dimension of Iron-Deficient Heart Failure.” European Journal of Heart Failure 26: 1631-1641. https://doi.org/10.1002/ejhf.3238

[10] Gong, Danni, Nianxuan Wu, Huan Chen, Weijie Zhang, Chenxi Yan, Chunlei Zhang, Yao Fu, and Hao Sun. 2024. “Phytic Acid-Loaded Polyvinyl Alcohol Hydrogel Promotes Wound Healing of Injured Corneal Epithelium Through Inhibiting Ferroptosis.” Redox Biology 76: 103354. https://doi.org/10.1016/j.redox.2024.103354

[11] Crack, Jason C., Basema K. Balasiny, Sophie P. Bennett, Matthew D. Rolfe, Afonso Froes, Fraser MacMillan, Jeffrey Green, Jeffrey A. Cole, and Nick E. Le Brun. 2022. “The Di-Iron Protein YtfE Is a Nitric Oxide-Generating Nitrite Reductase Involved in the Management of Nitrosative Stress.” Journal of the American Chemical Society 144: 7129-7145. https://doi.org/10.1021/jacs.1c12407

[12] Wu, Xintong, Zhonghong Zhang, Xiao Zhang, Yaping Guo, Feng Liu, Jianwei Gong, Li Li, Xinyu Chen, and Zhipeng Li. 2023. “Upregulation of A20 and TAX1BP1 Contributes to the Anti-Neuroinflammatory and Antidepressant Effects of Bavachalcone.” International Immunopharmacology 122: 110552. https://doi.org/10.1016/j.intimp.2023.110552

[13] Liu, Zi-bo, Xin-yun Fan, Chen-wei Wang, Xun Ye, and Chun-jie Wu. 2023. “Potentially Active Compounds That Improve PAD Through Angiogenesis: A Review.” Biomedicine & Pharmacotherapy 168: 115634. https://doi.org/10.1016/j.biopha.2023.115634

[14] Qian, Yun, Yang Yang, Kai Wang, Wenjun Zhou, Yanqi Dang, Mingzhe Zhu, Fenghua Li, and Guang Ji. 2019. “2′-Hydroxychalcone Induced Cytotoxicity via Oxidative Stress in the Lipid-Loaded Hepg2 Cells.” Frontiers in Pharmacology 10: 2019. https://doi.org/10.3389/fphar.2019.01390

[15] Singh, Surya, Gopal Pathuri, Adam Asch, Chinthalapally Rao, and Venkateshwar Madka. 2024. “Stat3 Inhibitors TTI-101 and SH5-07 Suppress Bladder Cancer Cell Survival in 3D Tumor Models.” Cells 13: 1463. https://www.mdpi.com/2073-4409/13/17/1463

[16] Zhang, Yueteng, Junjie Zhang, Menglong Li, Yan Qiao, Wei Wang, Lu Ma, and Kangdong Liu. 2023. “Target Discovery of Bioactive Natural Products With Native-Compound-Coupled CNBr-Activated Sepharose 4B Beads (NCCB): Applications, Mechanisms and Outlooks.” Bioorganic & Medicinal Chemistry 96: 117483. https://doi.org/10.1016/j.bmc.2023.117483

[17] Molina, Daniel Martinez, Rozbeh Jafari, Marina Ignatushchenko, Takahiro Seki, E. Andreas Larsson, Chen Dan, Lekshmy Sreekumar, Yihai Cao, and Pär Nordlund. 2013. “Monitoring Drug Target Engagement in Cells and Tissues Using the Cellular Thermal Shift Assay.” Science 341: 84-87. https://doi.org/10.1126/science.1233606

[18] Guo, Qian, Christopher Qian, Xinyu Wang, and Zhong-Ming Qian. 2025. “Transferrin Receptors.” Experimental & Molecular Medicine 57: 724-732. https://doi.org/10.1038/s12276-025-01436-x

[19] Gao, Yingbo, Boda Wang, Mengrui Hu, Yuhan Ma, and Bin Zheng. 2024. “The Role of Iron in Atherosclerosis and Its Association With Related Diseases.” Current Atherosclerosis Reports 27(1): 1. https://doi.org/10.1007/s11883-024-01251-1

[20] Jian, Jinlong, Qing Yang, and Xi Huang. 2011. “Src Regulates Tyr20 Phosphorylation of Transferrin Receptor-1 and Potentiates Breast Cancer Cell Survival *.” Journal of Biological Chemistry 286: 35708-35715. https://doi.org/10.1074/jbc.M111.271585

[21] Radin, Daniel P., and Parth Patel. 2016. “Delineating the Molecular Mechanisms of Tamoxifen's Oncolytic Actions in Estrogen Receptor-Negative Cancers.” European Journal of Pharmacology 781: 173-180. https://doi.org/10.1016/j.ejphar.2016.04.017

[22] Wang, Biao, Jiqin Zhang, Fei Song, Mi Tian, Bizhi Shi, Hua Jiang, Wen Xu, et al. 2016. “EGFR Regulates Iron Homeostasis to Promote Cancer Growth Through Redistribution of Transferrin Receptor 1.” Cancer Letters 381: 331-340. https://doi.org/10.1016/j.canlet.2016.08.006

[23] Shen, Ying, Xin Li, Bin Zhao, Yanru Xue, Shenghang Wang, Xin Chen, Jiancheng Yang, Huanhuan Lv, and Peng Shang. 2018. “Iron Metabolism Gene Expression and Prognostic Features of Hepatocellular Carcinoma.” Journal of Cellular Biochemistry 119: 9178-9204. https://doi.org/10.1002/jcb.27184

[24] Tran, Phong, Pradeep Mishra, Leonard G. Williams, Roman Moskalenko, Sushma Sharma, Anna Karin Nilsson, Danielle L. Watt, et al. 2024. “Altered dNTP Pools Accelerate Tumor Formation in Mice.” Nucleic Acids Research 52: 12475-12486. https://doi.org/10.1093/nar/gkae843

[25] Patel, Dillum, Alison L. Dickson, Greice M. Zickuhr, In Hwa Um, Oliver J. Read, Clarissa M. Czekster, Peter Mullen, David J. Harrison, and Jennifer Bré. 2024. “Defining the Mode of Action of Cisplatin Combined With NUC-1031, a Phosphoramidate Modification of Gemcitabine.” Translational Oncology 50: 102114. https://doi.org/10.1016/j.tranon.2024.102114

[26] Dai, Yuhe, Qianbo Zhang, Rifang Gu, Jitao Chen, Penghui Ye, Huan Zhu, Ming Tang, and Xuqiang Nie. 2024. “Metal Ion Formulations for Diabetic Wound Healing: Mechanisms and Therapeutic Potential.” International Journal of Pharmaceutics 667: 124889. https://doi.org/10.1016/j.ijpharm.2024.124889

[27] Bolan, Shiv, Hasintha Wijesekara, Achali Ireshika, Tao Zhang, Mingjun Pu, Gianniantonio Petruzzelli, Francesca Pedron, et al. 2023. “Tungsten Contamination, Behavior and Remediation in Complex Environmental Settings.” Environment International 181: 108276. https://doi.org/10.1016/j.envint.2023.108276

[28] Silvestri, Laura, Mariateresa Pettinato, Valeria Furiosi, Letizia Bavuso Volpe, Antonella Nai, and Alessia Pagani. 2023. “Managing the Dual Nature of Iron to Preserve Health.” International Journal of Molecular Sciences 24: 3995. https://doi.org/10.3390/ijms24043995. https://www.mdpi.com/1422-0067/24/4/3995

[29] Zhang, Chenliang, Tingting Huang, and Liping Li. 2024. “Targeting Cuproptosis for Cancer Therapy: Mechanistic Insights and Clinical Perspectives.” Journal of Hematology & Oncology 17: 68. https://doi.org/10.1186/s13045-024-01589-8

[30] Rossetto, Daniele, Nemanja Cvjetan, Peter Walde, and Sheref S. Mansy. 2024. “Protocellular Heme and Iron-Sulfur Clusters.” Accounts of Chemical Research 57: 2293-2302. https://doi.org/10.1021/acs.accounts.4c00254

[31] Dancis, Andrew, Ashutosh K. Pandey, and Debkumar Pain. 2024. “Mitochondria Function in Cytoplasmic FeS Protein Biogenesis.” Biochimica et Biophysica Acta (BBA) - Molecular Cell Research 1871: 119733. https://doi.org/10.1016/j.bbamcr.2024.119733

[32] Nagae, Fritz, Yasuto Murayama, and Tsuyoshi Terakawa. 2024. “Molecular Mechanism of Parental H3/H4 Recycling At a Replication Fork.” Nature Communications 15: 9485. https://doi.org/10.1038/s41467-024-53187-4

[33] Foskolou, Iosifina P., Christian Jorgensen, Katarzyna B. Leszczynska, Monica M. Olcina, Hanna Tarhonskaya, Bauke Haisma, Vincenzo D'Angiolella, et al. 2017. “Ribonucleotide Reductase Requires Subunit Switching in Hypoxia to Maintain DNA Replication.” Molecular Cell 66: 206-220.e9. https://doi.org/10.1016/j.molcel.2017.03.005

[34] Zangouei, Amir Sadra, Malihe Zangoue, Negin Taghehchian, Alireza Zangooie, Hamid Reza Rahimi, Ehsan Saburi, Mahya Sadat Alavi, and Meysam Moghbeli. 2023. “Cell Cycle Related Long Non-Coding RNAs as the Critical Regulators of Breast Cancer Progression and Metastasis.” Biological Research 56(1): 1. https://doi.org/10.1186/s40659-022-00411-4

[35] Hsu, Jocelyn Y., Nathan D. Seligson, John L. Hays, Wayne O. Miles, and James L. Chen. 2022. “Clinical Utility of CDK4/6 Inhibitors in Sarcoma: Successes and Future Challenges.” JCO Precision Oncology 6: e2100211. https://doi.org/10.1200/po.21.00211

RIGHTS & PERMISSIONS

2025 The Author(s). iMeta published by John Wiley & Sons Australia, Ltd on behalf of iMeta Science.