Abstract
Objective Gallbladder cancer (GBC) poses a significant health burden with dismal prognosis due to frequent metastasis and recurrence. Although evodiamine has demonstrated potent inhibitory effects on proliferation and metastasis in various cancers, its role in GBC remains unexplored, and the underlying mechanisms are yet to be elucidated. Methods: The inhibitory effects of evodiamine on GBC cells were evaluated in vitro using a cell viability assay. Cell migration capacity was assessed via wound healing and Transwell assays. Apoptosis and cell cycle distribution were analyzed by flow cytometry and Western blotting (WB). The molecular mechanisms were investigated using Quantitative polymerase chain reaction (qPCR) and WB to quantify ZEB1 gene expression. In vivo, the anti-tumor activity of evodiamine was verified in a nude mouse model.Results: Evodiamine significantly inhibited the proliferation of GBC cells. Flow cytometry and Western blotting revealed that evodiamine induced G2/M phase arrest and promoted apoptosis. mRNA-sequencing (mRNA-seq) demonstrated that evodiamine suppressed the transcription of ZEB1 and genes in the PI3K-Akt signaling pathway. Consistent with in vitro findings, evodiamine exhibited remarkable antitumor effects in a nude mouse model. Conclusion: This study confirms that evodiamine inhibits GBC cell proliferation and induces apoptosis. The mechanism involves suppression of ZEB1 expression and inactivation of the PI3K-Akt signaling pathway.
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The datasets generated and analysed during the current study are available in the GEO repository, GSE312961.
References
Zhao, C. et al. Ponicidin inhibited gallbladder cancer proliferation and metastasis by decreasing MAGEB2 expression through FOXO4. Phytomedicine 114, 154785 (2023).
Geng, Y. et al. Long-term exposure to genistein inhibits the proliferation of gallbladder cancer by downregulating the MCM complex. Sci. Bull. 67, 813–824 (2022).
Yang, Z. et al. Successful conversion surgery for locally advanced gallbladder cancer after gemcitabine and nab-paclitaxel chemotherapy. Front. Oncol. 12, 977963 (2022).
Cui, X. Y. et al. Modified FOLFIRINOX for unresectable locally advanced or metastatic gallbladder cancer, a comparison with GEMOX regimen. Hepatobiliary Surg. Nutr. 10, 498–506 (2021).
Liu, A. J. et al. Evodiamine, a plant alkaloid, induces calcium/JNK-mediated autophagy and calcium/mitochondria-mediated apoptosis in human glioblastoma cells. Chemico-Biol. Interact. 205, 20–28 (2013).
Sun, Q., Xie, L., Song, J., Li, X. & Evodiamine A review of its pharmacology, toxicity, pharmacokinetics and Preparation researches. J. Ethnopharmacol. 262, 113164 (2020).
Hu, X. et al. Antiproliferative effects of alkaloid Evodiamine and its derivatives. Int. J. Mol. Sci. 19, 3403 (2018).
Guo, X. X. et al. Evodiamine induces apoptosis in SMMC-7721 and HepG2 cells by suppressing NOD1 signal pathway. Int. J. Mol. Sci. 19, 3419 (2018).
Solanki, R., Rajput, P. K., Jodha, B., Yadav, U. C. S. & Patel, S. Enhancing apoptosis-mediated anticancer activity of Evodiamine through protein-based nanoparticles in breast cancer cells. Sci. Rep. 14, 2595 (2024).
Jiang, Z. B. et al. Evodiamine suppresses non-small cell lung cancer by elevating CD8 + T cells and downregulating the MUC1-C/PD-L1 axis. J. Exp. Clin. Cancer Res. 39, 249 (2020).
Li, L. et al. HN1L-mediated transcriptional axis AP-2γ/METTL13/TCF3-ZEB1 drives tumor growth and metastasis in hepatocellular carcinoma. Cell. Death Differ. 26, 2268–2283 (2019).
Sheng, W. et al. Musashi2 promotes EGF-induced EMT in pancreatic cancer via ZEB1-ERK/MAPK signaling. J. Exp. Clin. Cancer Res. 39, 16 (2020).
Zhang, D. et al. LINC01189-miR-586-ZEB1 feedback loop regulates breast cancer progression through Wnt/β-catenin signaling pathway. Mol. Ther. Nucleic Acids. 25, 455–467 (2021).
Liu, M. et al. Zinc-Dependent regulation of ZEB1 and YAP1 coactivation promotes Epithelial-Mesenchymal transition plasticity and metastasis in pancreatic cancer. Gastroenterology 160, 1771–1783e1 (2021).
Pastushenko, I. & Blanpain, C. EMT transition States during tumor progression and metastasis. Trends Cell Biol. 29, 212–226 (2019).
Jeng, K. S. et al. Glioma-Associated oncogene homolog inhibitors have the potential of suppressing cancer stem cells of breast cancer. Int. J. Mol. Sci. 19, 1375 (2018).
Jiang, M. et al. miR-1254 inhibits cell proliferation, migration, and invasion by down-regulating Smurf1 in gastric cancer. Cell. Death Dis. 10, 32 (2019).
Glaviano, A. et al. PI3K/AKT/mTOR signaling transduction pathway and targeted therapies in cancer. Mol. Cancer. 22, 138 (2023).
Li, L. et al. Research Advances in Antitumor Mechanism of Evodiamine. Journal of Chemistry 2784257 (2022).
Zhu, B. et al. Induction of phosphatase shatterproof 2 by Evodiamine suppresses the proliferation and invasion of human cholangiocarcinoma. Int. J. Biochem. Cell. Biol. 108, 98–110 (2019).
Zhao, S. et al. Evodiamine inhibits proliferation and promotes apoptosis of hepatocellular carcinoma cells via the Hippo-Yes-Associated protein signaling pathway. Life Sci. 251, 117424 (2020).
Wu, W. S., Chien, C. C., Liu, K. H., Chen, Y. C. & Chiu, W. T. Evodiamine prevents glioma Growth, induces glioblastoma cell apoptosis and cell cycle arrest through JNK activation. Am. J. Chin. Med. 45, 879–899 (2017).
Zhang, L. et al. Evodiamine inhibits ESCC by inducing M-phase cell-cycle arrest via CUL4A/p53/p21 axis and activating noxa-dependent intrinsic and DR4-dependent extrinsic apoptosis. Phytomedicine 108, 154493 (2023).
Turajlic, S. & Swanton, C. Metastasis as an evolutionary process. Science 352, 169–175 (2016).
Mittal, V. Epithelial mesenchymal transition in tumor metastasis. Annu. Rev. Pathol. 13, 395–412 (2018).
Drápela, S., Bouchal, J., Jolly, M. K., Culig, Z. & Souček, K. ZEB1: A critical regulator of cell Plasticity, DNA damage Response, and therapy resistance. Front. Mol. Biosci. 7, 36 (2020).
Zhou, Y. et al. ZEB1 enhances Warburg effect to facilitate tumorigenesis and metastasis of HCC by transcriptionally activating PFKM. Theranostics 11, 5926–5938 (2021).
Li, L. et al. ZEB1 serves as an oncogene in acute myeloid leukaemia via regulating the PTEN/PI3K/AKT signalling pathway by combining with P53. J. Cell. Mol. Med. 25, 5295–5304 (2021).
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Funding
This research was supported by Science and Technology Innovation Project of Shanghai Putuo District Health and Wellness Plan (No. ptkwws202308), Shanghai Health System Key Disciplines - General Surgery (No. 2024ZDXK0046),the Clinical Specialized Disease Construction Project of Shanghai Putuo District Municipal Health Commission (No.2024tszb01༉, Fujian Provincial Natural Science Foundation of China (No. 2024J01222) and the National Key Research and Development Program of China (No. 2024YFA1108604).
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Wei Li and Yafeng Chen designed the experiments. Yijie Li conducted the experiments and wrote the manuscript. Shufen Zhou, Hanzheng Xu, Hongye Zhou, Kunqi Sun, and Siyang Hu assisted with references collection. Jiahua Yang and Ke Xu reviewed and revised the manuscript. All authors have read and approved the final manuscript.
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This study adhered strictly to the ARRIVE guidelines in its design and reporting to ensure animal welfare and research transparency. All experiments were conducted in compliance with the ethical review requirements of [the Institutional Animal Care and Use Committee of Putuo Hospital, Shanghai University of Traditional Chinese Medicine, China] (Ethics Approval Number: DWEC-A-2023-01-1–69), and efforts were made to minimize the number of animals used and their suffering.This study did not involve human participants, human experimentation, or the use of human tissue samples. Therefore, no institutional review board or ethics committee approval was required or sought.
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Li, Y., Zhou, S., Xu, H. et al. Alkaloids from Evodia rutaecarpa inhibit the occurrence and development of gallbladder cancer in vivo and in vitro. Sci Rep (2026). https://doi.org/10.1038/s41598-026-35563-w
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DOI: https://doi.org/10.1038/s41598-026-35563-w
