Tongguanteng injection induces cell cycle arrest and drives ferroptosis through AURKA/KEAP1/NRF2 axis in breast cancer

  • Phytomedicine. 2026 Jul:156:158250. doi: 10.1016/j.phymed.2026.158250.
Hanlu Liang  1 ,  Qixuan Ding  2 ,  Jiayi Chen  1 ,  Shuai Zhao  1 ,  Yangyun Zhou  2 ,  Yujie Hu  2 ,  Lingyan Xu  2 ,  Meizhi Shi  2 ,  Jiao Yang  2 ,  Xiangqi Zhang  2 ,  Jingxian Zhang  2 ,  Cheng Guo  3 ,  Yonglong Han  4 ,  Junjun Chen  5
Affiliations
  • 1. Department of Pharmacy, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China; College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China.
  • 2. Department of Pharmacy, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China.
  • 3. Department of Pharmacy, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China. Electronic address: [email protected].
  • 4. Department of Pharmacy, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China; College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China. Electronic address: [email protected].
  • 5. Department of Pharmacy, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China. Electronic address: [email protected].
PMID: 42105703 DOI: 10.1016/j.phymed.2026.158250
Abstract

Background: Tongguanteng Injection (TGT), derived from the stems of Marsdenia tenacissima, has been demonstrated to exert anti-tumor effects. Aurora Kinase A (AURKA) is a key regulator of cell cycle progression and has become a promising target for breast Cancer treatment. However, the role of AURKA in Ferroptosis is unclear, particularly regarding its contribution to the therapeutic effect of TGT in breast Cancer.

Methods: The chemical components of TGT were analyzed by Ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). The anti-breast Cancer effect of TGT were detected in vitro and in vivo. RNA-seq analysis were used to identify the targets of TGT treatment. Subsequently, flow cytometry, RT-PCR, western blot, Co-IP, immunofluorescence and immunohistochemistry were employed to elucidate the regulatory mechanisms of TGT on Ferroptosis and cell cycle. Concurrently, Ferroptosis and cell cycle associated pathways were investigated in the plasmids and siRNAs transfected cells. Furthermore, tumor bearing mouse model was established by 4T1 cells stably transduced with lentivirus for further validating the findings.

Results: A total of 136 compounds were identified from TGT. The study demonstrates that TGT exerts an anti-breast Cancer effect by inducing G2/M phase arrest and Ferroptosis, evidenced by downregulation of CDK1 and Cyclin B, as well as increased Fe2+ accumulation, glutathione (GSH) depletion, and elevated lipid peroxidation. RNA-seq analysis revealed that AURKA was significantly downregulated and identified as a key target of TGT treatment. Mechanistically, TGT disrupts the AURKA-Kelch-like ECH-associated protein 1 (KEAP1) interaction, leading to nuclear factor E2-related factor 2 (NRF2) nuclear translocation and subsequent heme oxygenase 1 (HO-1)-mediated Ferroptosis in vitro. Knockdown of AURKA or treatment with its inhibitor alisertib both induce G2/M phase arrest and Ferroptosis, while AURKA overexpression reverses the effects of TGT and accelerates tumor progression. Combining TGT with docetaxel enhances Ferroptosis sensitivity and amplifies the regulatory effect of AURKA on cell cycle and Ferroptosis. Consistent with in vitro results, TGT significantly inhibited tumor growth, arrested cell cycle and promoted Ferroptosis in 4T1-AURKALV cell bearing mice.

Conclusion: These findings propose that TGT targets AURKA/KEAP1/NRF2 axis to exert anti-breast Cancer effect, and AURKA inhibition represents a potential strategy for breast Cancer treatment through inducing cell cycle arrest and driving Ferroptosis.

Keywords
AURKA; Breast cancer; Cell cycle; Ferroptosis; Tongguanteng injection.
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