Brevilin A induces mitochondrial dysfunction and endoplasmic reticulum stress in breast cancer through WWP2-mediated KLF5 degradation

i

This article has been updated. A Correction to this article was published on .

i

The Original Article was published on

i

The Original Article was published on

i

This article has been updated

i

This article was retracted on

i

This article has been updated

Abstract

Breast cancer (BC) remains a significant challenge to women’s health, and identifying effective molecular targets is crucial for therapeutic advancement. Krüppel-like factor 5 (KLF5), an oncogenic transcription factor, is highly expressed in BC, yet its development as a therapeutic target has been hindered by its intrinsically disordered structure. In this study, we identified Brevilin A (BrA), a natural sesquiterpenoid fromCentipeda minima, as a potent KLF5 degrader. We demonstrated that BrA directly binds to KLF5 and promotes its ubiquitin-mediated degradation through the E3 ligase WWP2. BrA significantly inhibited BC cell proliferation and induced severe mitochondrial dysfunction and endoplasmic reticulum (ER) stress, as evidenced by reduced mitochondrial respiration and activation of the PERK-EIF2α pathway. Mechanistically, BrA enhanced the recruitment of WWP2 to KLF5, leading to its polyubiquitination and subsequent proteasomal degradation. KLF5 overexpression effectively rescued the mitochondrial and ER homeostasis disrupted by BrA, whereas KLF5 knockdown rendered cells insensitive to BrA-induced cytotoxicity. In vivo studies using zebrafish and nude mouse xenograft models confirmed that BrA suppressed tumor growth by downregulating KLF5 and its downstream metabolic effectors. Collectively, our findings reveal that BrA exerts antitumor effects by promoting WWP2-mediated ubiquitination and degradation of KLF5, thereby disrupting mitochondrial and ER integrity, highlighting its potential as a natural lead compound for BC treatment.

Keywords

Brevilin A / Breast cancer / KLF5 / Mitochondrial dysfunction / Endoplasmic reticulum stress / WWP2

CHANGE HISTORY

A Correction to this paper has been published:
https://doi.org

The Retraction Note was updated to reflect reception of statements of disagreement from all authors.

This article has been retracted.Please see the Retraction Notice for more detail:
https://doi.org

References

[1] Bray F, Laversanne M, Sung H, et al. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2024; 74(3):229-263. https://doi.org/10.3322/caac.21834.

[2] Yang R, Li Y, Wang H, et al. Therapeutic progress and challenges for triple negative breast cancer: targeted therapy and immunotherapy. Mol Biomed. 2022; 3(1):8. https://doi.org/10.1186/s43556-022-00071-6.

[3] Yuce K, Ozkan AI . The kruppel-like factor (KLF) family, diseases, and physiological events. Gene. 2024; 895:148027. https://doi.org/10.1016/j.gene.2023.148027.

[4] Gao Y, Ding Y, Chen H, et al. Targeting Krüppel-like factor 5 (KLF5) for cancer therapy. Curr Top Med Chem. 2015; 15(8):699-713. https://doi.org/10.2174/1568026615666150302105052.

[5] Yang S, Feng T, Li H . KLF5, a novel therapeutic target in squamous cell carcinoma. DNA Cell Biol. 2021; 40(12):1503-1512. https://doi.org/10.1089/dna.2021.0674.

[6] Ekstrom TL, Dai Z, Thiel J, et al. KLF5 controls subtype-independent highly interactive enhancers in pancreatic cancer to regulate cell survival. Sci Adv. 2026;12(12):eaea2106. https://doi.org/10.1126/sciadv.aea2106.

[7] Zeng J, Li Q, Zhang J, et al. KLF5 activation promotes malignant transformation to drive development of lung squamous cell carcinoma. Cancer Res.. 2026; 86(7):1622-1638. https://doi.org/10.1158/0008-5472.CAN-25-2139.

[8] Luo Y, Chen C . The roles and regulation of the KLF5 transcription factor in cancers. Cancer Sci. 2021; 112(6):2097-2117. https://doi.org/10.1111/cas.14910.

[9] Tang Y, Liu R, Zhu J, et al. Positive feedback regulation between KLF5 and XPO1 promotes cell cycle progression of basal like breast cancer. Adv Sci (Weinh). 2025; 12(16):e2412096. https://doi.org/10.1002/advs.202412096.

[10] Li J, Liang Y, Zhou S, et al. UCHL1 contributes to insensitivity to endocrine therapy in triple-negative breast cancer by deubiquitinating and stabilizing KLF5. Breast Cancer Res.. 2024; 26(1):44. https://doi.org/10.1186/s13058-024-01800-1.

[11] Xie B, Wang M, Xiao Y, et al. Tryptophan metabolic gatekeeping in epithelial repair: GPR35-KLF5 circuitry decodes mucosal damage signals for repair programming. Cell Death Dis. 2026; 17(1):25. https://doi.org/10.1038/s41419-025-08237-0.

[12] Zhang X, Liu F, Wang Q, et al. Overexpressed microRNA-506 and microRNA-124 alleviate H₂O₂-induced human cardiomyocyte dysfunction by targeting krüppel-like factor 4/5. Mol Med Rep. 2017; 16(4):5363-5369. https://doi.org/10.3892/mmr.2017.7243.

[13] Li LY, Yang Q, Jiang YY, et al. Interplay and cooperation between SREBF1 and master transcription factors regulate lipid metabolism and tumor-promoting pathways in squamous cancer. Nat Commun. 2021; 12(1):4362. https://doi.org/10.1038/s41467-021-24656-x.

[14] Zhang Y, Chen JC, Zheng JH, et al. Pterosin B improves cognitive dysfunction by promoting microglia M1/M2 polarization through inhibiting Klf5/Parp14 pathway. Phytomedicine. 2024; 135:156152. https://doi.org/10.1016/j.phymed.2024.156152.

[15] Ma D, Zheng B, Liu HL, et al. Klf5 down-regulation induces vascular senescence through eIF5a depletion and mitochondrial fission. PLoS Biol. 2020; 18(8):e3000808. https://doi.org/10.1371/journal.pbio.3000808.

[16] Gao L, Yang X, Li Y, et al. Curcumol inhibits KLF5-dependent angiogenesis by blocking the ROS/ERK signaling in liver sinusoidal endothelial cells. Life Sci.. 2021; 264:118696. https://doi.org/10.1016/j.lfs.2020.118696.

[17] Ma L, Lin C, Xiao Z, et al. Acetylated KLF5 inhibits LACTB transcription, mediates mitochondrial dynamics, and regulates colorectal cancer cell stemness and differentiation. Int J Biol Macromol. 2025; 333(Pt 2):148905. https://doi.org/10.1016/j.ijbiomac.2025.148905.

[18] Deng J, Wang D, Jiang K, et al. Targeting PDK4 to mitigate osimertinib-induced cardiotoxicity: insights into mitochondria-endoplasmic reticulum crosstalk and necroptosis. Free Radic Biol Med. 2025; 240:267-283. https://doi.org/10.1016/j.freeradbiomed.2025.08.017.

[19] Liu Y, Mao ZH, Huang J, et al. Mitochondria-associated endoplasmic reticulum membranes in human health and diseases. MedComm (2020). 2025; 6(7):e70259. https://doi.org/10.1002/mco2.70259.

[20] Alizade A, Evyapan G, Celik IS, et al. Metformin induces mitochondria-mediated and endoplasmic reticulum stress-mediated apoptosis and inhibits angiogenesis-related gene expression in breast cancer cells via targeting VEGF-A/VEGFR2/NRP1. Croat Med J. 2025; 66(2):115-124. https://doi.org/10.3325/cmj.2025.66.115.

[21] Chen C, Sun X, Ran Q, et al. Ubiquitin-proteasome degradation of KLF5 transcription factor in cancer and untransformed epithelial cells. Oncogene. 2005; 24(20):3319-3327. https://doi.org/10.1038/sj.onc.1208497.

[22] Wang TT, Zhang LL, Li FB, et al. LN-439A, a novel BAP1 inhibitor, suppresses the growth of basal-like breast cancer by degrading KLF5. Acta Pharmacol Sin. 2025; 46(3):715-727. https://doi.org/10.1038/s41401-024-01361-1.

[23] Zhao D, Zhi X, Zhou Z, et al. TAZ antagonizes the WWP1-mediated KLF5 degradation and promotes breast cell proliferation and tumorigenesis. Carcinogenesis. 2012; 33(1):59-67. https://doi.org/10.1093/carcin/bgr242.

[24] Shi P, Liu W, Tala, et al. Metformin suppresses triple-negative breast cancer stem cells by targeting KLF5 for degradation. Cell Discov. 2017; 3:17010. https://doi.org/10.1038/celldisc.2017.10.

[25] Du JX, Hagos EG, Nandan MO, et al. The E3 ubiquitin ligase SMAD ubiquitination regulatory factor 2 negatively regulates Krüppel-like factor 5 protein. J Biol Chem. 2011; 286(46):40354-40364. https://doi.org/10.1074/jbc.M111.258707.

[26] Zhao KW, Sikriwal D, Dong X, et al. Oestrogen causes degradation of KLF5 by inducing the E3 ubiquitin ligase EFP in ER-positive breast cancer cells. Biochem J. 2011; 437(2):323-333. https://doi.org/10.1042/BJ20101388.

[27] Wan F, Qian C, Liu X, et al. Sculponeatin A induces mitochondrial dysfunction in non-small cell lung cancer through WWP2-mediated degradation of mitochondrial STAT3. Br J Pharmacol. 2025; 182(12):2662-2681. https://doi.org/10.1111/bph.17460.

[28] Zhang Z, Lu YX, Liu F, et al. lncRNA BREA2 promotes metastasis by disrupting the WWP2-mediated ubiquitination of Notch1. Proc Natl Acad Sci U S A. 2023; 120(8):e2206694120. https://doi.org/10.1073/pnas.2206694120.

[29] Chan CO, Xie XJ, Wan SW, et al. Qualitative and quantitative analysis of sesquiterpene lactones in Centipeda minima by UPLC-Orbitrap-MS & UPLC-QQQ-MS. J Pharm Biomed Anal. 2019; 174:360-366. https://doi.org/10.1016/j.jpba.2019.05.067.

[30] Liu YQ, Zhou GB . Promising anticancer activities and mechanisms of action of active compounds from the medicinal herb Centipeda minima (L.) A. Braun & Asch. Phytomedicine. 2022; 106:154397. https://doi.org/10.1016/j.phymed.2022.154397.

[31] Qu Z, Lin Y, Mok DK, et al. Brevilin A, a natural sesquiterpene lactone inhibited the growth of triple-negative breast cancer cells via Akt/mTOR and STAT3 signaling pathways. Onco Targets Ther. 2020; 13:5363-5373. https://doi.org/10.2147/OTT.S256833.

[32] Saleem MZ, Nisar MA, Alshwmi M, et al. Brevilin A inhibits STAT3 signaling and induces ROS-dependent apoptosis, mitochondrial stress and endoplasmic reticulum stress in MCF-7 breast cancer cells. Onco Targets Ther. 2020; 13:435-450. https://doi.org/10.2147/OTT.S228702.

[33] Xiang Y, Wan F, Ren Y, et al. Polyphyllin VII induces autophagy-dependent ferroptosis in human gastric cancer through targeting T-lymphokine-activated killer cell-originated protein kinase. Phytotherapy Research. 2023; 37(12):5803-5820. https://doi.org/10.1002/ptr.7986.

[34] Peng P, Ren Y, Wan F, et al. Sculponeatin A promotes the ETS1-SYVN1 interaction to induce SLC7A11/xCT-dependent ferroptosis in breast cancer. Phytomedicine. 2023; 117:154921. https://doi.org/10.1016/j.phymed.2023.154921.

[35] Ren Y, Chen J, Zhan X, et al. Liquid-liquid phase separation of GPS2-LATS1 promotes colorectal cancer progression by reprogramming lipid metabolism. Oncogene. 2025; 44(39):3741-3754. https://doi.org/10.1038/s41388-025-03498-7.

[36] Si Y, Ou H, Jin X, et al. G protein pathway suppressor 2 suppresses aerobic glycolysis through RACK1-mediated HIF-1α degradation in breast cancer. Free Radic Biol Med. 2024; 222:478-492. https://doi.org/10.1016/j.freeradbiomed.2024.06.021.

[37] Xiang YC, Shen J, Si Y, et al. Paris saponin VII, a direct activator of AMPK, induces autophagy and exhibits therapeutic potential in non-small-cell lung cancer. Chin J Nat Med. 2021; 19(3):195-204. https://doi.org/10.1016/S1875-5364(21)60021-3.

[38] Wu Y, Si Y, Xiang Y, et al. Polyphyllin I activates AMPK to suppress the growth of non-small-cell lung cancer via induction of autophagy. Arch Biochem Biophys. 2020; 687:108285. https://doi.org/10.1016/j.abb.2020.108285.

[39] Xiang YC, Peng P, Liu XW, et al. Paris saponin VII, a Hippo pathway activator, induces autophagy and exhibits therapeutic potential against human breast cancer cells. Acta Pharmacol Sin. 2022; 43(6):1568-1580. https://doi.org/10.1038/s41401-021-00755-9.

[40] Xu J, Chen Y, Yang R, et al. Cucurbitacin B inhibits gastric cancer progression by suppressing STAT3 activity. Arch Biochem Biophys. 2020; 684:108314. https://doi.org/10.1016/j.abb.2020.108314.

[41] Si Y, Zhang H, Peng P, et al. G protein pathway suppressor 2 suppresses gastric cancer by destabilizing epidermal growth factor receptor. Cancer Sci.. 2021; 112(12):4867-4882. https://doi.org/10.1111/cas.15151.

[42] Ren Y, Wu H, Tan M, et al. Acetylation of MOB1 mediates polyphyllin II-reduced lysosome biogenesis in breast cancer by promoting the cytoplasmic retention of the YAP/TFEB coactivator complex. Phytomedicine. 2024; 122:155152. https://doi.org/10.1016/j.phymed.2023.155152.

[43] Dittmar G, Winklhofer KF . Linear ubiquitin chains: cellular functions and strategies for detection and quantification. Front Chem. 2019; 7:915. https://doi.org/10.3389/fchem.2019.00915.

[44] Ziegler DV, Parashar K, Leal-Esteban L, et al. CDK4 inactivation inhibits apoptosis via mitochondria-ER contact remodeling in triple-negative breast cancer. Nat Commun. 2025; 16(1):541. https://doi.org/10.1038/s41467-024-55605-z.

[45] Sassano ML, Tyurina YY, Diokmetzidou A, et al. Endoplasmic reticulum-mitochondria contacts are prime hotspots of phospholipid peroxidation driving ferroptosis. Nat Cell Biol. 2025; 27(6):902-917. https://doi.org/10.1038/s41556-025-01668-z.

[46] Urra H, Dufey E, Avril T, et al. Endoplasmic reticulum stress and the hallmarks of cancer. Trends Cancer. 2016; 2(5):252-262. https://doi.org/10.1016/j.trecan.2016.03.007.

[47] Chen J, Ying K, Sun J, et al. NEDD4L affects KLF5 stability through ubiquitination to control ferroptosis and radiotherapy resistance in oesophageal squamous cell carcinoma. J Cell Mol Med. 2024; 28(18):e70062. https://doi.org/10.1111/jcmm.70062.