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Ginsenoside (20)S-APPT induces ferroptosis in hepatocellular carcinoma and cholangiocarcinoma by targeting FSP1

Acta Pharmacologica Sinica volume 46, pages 3273–3290 (2025)Cite this article

Abstract

Triggering ferroptosis has recently been recognized as a promising approach for cancer treatment. However, current ferroptosis inducers, such as glutathione peroxidase 4 (GPX4) inhibitors, face limitations in terms of druggability and safety. In this study, we performed a phenotypic screen of a 180-compound natural product library and identified (20S)-protopanaxatriol ((20)S-APPT), a ginsenoside derivative, as a potent ferroptosis inducer with a favorable safety profile both in vitro and in vivo. We demonstrated that (20)S-APPT induced ferroptosis by targeting the plasma membrane-localized CoQ10 oxidoreductase FSP1. FSP1 inhibition promoted ACSL4-dependent arachidonic acid oxidation and mitochondrial ROS production, thereby increasing ferroptosis. Intriguingly, we revealed that FSP1 inhibition alone was sufficient to trigger ferroptosis in a subset of hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA) cells. Furthermore, the combined inhibition of FSP1 and γ-glutamylcysteine synthetase (GCS) synergistically induced ferroptosis in otherwise resistant cancer cells while sparing noncancerous cells. These results establish a previously unrecognized role for FSP1 in driving ferroptosis and highlight the therapeutic potential of cotargeting FSP1 and GCS in HCC and CCA.

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Fig. 1: (20)S-APPT is a novel ferroptosis inducer.
Fig. 2: SLC7A11-GSH pathway is dispensable for (20)S-APPT-induced ferroptosis.
Fig. 3: (20)S-APPT acts as an FSP1 inhibitor to induce ferroptosis.
Fig. 4: Plasma membrane-localized FSP1 plays a key role in (20)S-APPT-induced ferroptosis.
Fig. 5: FSP1 inhibition activates ACSL4 phosphorylation and promotes arachidonic acid oxidation.
Fig. 6: FSP1 inhibition induces mitochondrial ROS production to promote ferroptosis.
Fig. 7: The addiction of a subset of HCC and CCA cells to FSP1.
Fig. 8: GSH elimination through GCS inhibition effectively and broadly resensitizes cancer cells to FSP1 inhibitors.

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Data availability

The raw RNA sequencing data have been deposited in the Gene Expression Omnibus under the accession number GSE284669. All data in our study are available upon reasonable request from the corresponding authors.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 82173831), the Natural Science Foundation of China for Innovation Research Group (No. 81821005), the Shanghai Municipal Science and Technology Major Project, Foundation of Shanghai Science and Technology Committee (No. 21DZ2291100), Shandong Laboratory Program (No. SYS202205), “Personalized Medicines-Molecular Signature-based Drug Discovery and Development”, Strategic Priority Research Program of the Chinese Academy of Sciences (No. XDA12050102) and Key R&D Program of Shandong Province (2024CXPT029). We thank Dr. Ao Huang (Zhongshan Hospital, Fudan University) for kindly providing tumor samples. We are grateful to Dr. Chang-rong Ge (Karolinska Institute, Stockholm, Sweden) for his generous assistance in manuscript preparation and amendment.

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Author notes

  1. These authors contributed equally: Fan-yu Liu, Yuan-jie Yang.

Authors and Affiliations

  1. Division of Anti-tumor Pharmacology, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China

    Fan-yu Liu, Yuan-jie Yang, Yan Hong, Xiao-peng Ou-Yang, Min-min Zhang, Mei-yu Geng & Ai-jun Shen

  2. University of Chinese Academy of Sciences, Beijing, 100049, China

    Fan-yu Liu, Yuan-jie Yang & Mei-yu Geng

  3. Lingang Laboratory, Shanghai, 200031, China

    Xue-long Wang & Ai-jun Shen

  4. School of Life Science and Technology, ShanghaiTech University, Shanghai, 201210, China

    Xiao-peng Ou-Yang

  5. “Belt and Road” Joint Laboratory for Natural Products and Drug Discovery, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China

    Lu Chang, Nan-lin Zhu & Jia Liu

  6. Department of Liver Surgery and Transplantation, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Key Laboratory of Carcinogenesis and Cancer Invasion (Fudan University), Ministry of Education, Shanghai, 200032, China

    Ao Huang

  7. School of Pharmaceutical Science and Technology, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences, Hangzhou, 310058, China

    Jia Liu

  8. Shandong Laboratory of Yantai Drug Discovery, Bohai Rim Advanced Research Institute for Drug Discovery, Yantai, 264117, China

    Mei-yu Geng

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Contributions

FYL and AJS designed the study and prepared the manuscript. FYL and YJY performed the experiments and analyzed the experimental data. XLW provided bioinformatic supports. YH cultured and amplified PDX-derived primary tumor cells. LC, NLZ and JL performed metabolomic and lipidomic studies and analyzed data. AH provided tumor samples. XPOY and MMZ provided technical and material support. AJS, MYG, and JL supervised the study. All authors have read and approved the article.

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Correspondence to Jia Liu, Mei-yu Geng or Ai-jun Shen.

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Liu, Fy., Yang, Yj., Wang, Xl. et al. Ginsenoside (20)S-APPT induces ferroptosis in hepatocellular carcinoma and cholangiocarcinoma by targeting FSP1. Acta Pharmacol Sin 46, 3273–3290 (2025). https://doi.org/10.1038/s41401-025-01589-5

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