Journal of Shandong University (Health Sciences) ›› 2025, Vol. 63 ›› Issue (11): 8-17.doi: 10.6040/j.issn.1671-7554.0.2025.0294

• Preclinical Medicine • Previous Articles    

NEDD4L regulates ferroptosis by mediating ubiquitination of ALDOA in glioblastoma cells

NIE Qiucheng1,2, LI Yunfeng2, TIAN Jing3, LIU Xinjing1,2, SUN Lili1,2, WEI Yiju1,2   

  1. 1. College of Life Sciences, Shandong First Medical University, Taian 271016, Shandong, China;
    2. Medical Science and Technology Innovation Center, Shandong First Medical University, Jinan 250117, Shandong, China;
    3. Department of Blood Transfusion, Second Affiliated Hospital of Shandong First Medical University, Taian 271016, Shandong, China
  • Published:2025-11-28

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Abstract: Objective To investigate the role of the E3 ubiquitin ligase neural precursor cell expressed developmentally down-regulated 4 like(NEDD4L)-mediating ferroptosis in glioblastoma, aiming to elucidate molecular mechanism of NEDD4L-mediated ubiquitination of aldolase A(ALDOA)which regulates ferroptosis pathway. Methods The effects of knockdown of NEDD4L on ferroptosis in glioblastoma(GBM)were assessed through cell death and lipid peroxidation assays. ALDOA was identified as a potential substrate modified by NEDD4L using protein Western blotting and co-immunoprecipitation(Co-IP)techniques. The impact of NEDD4L or ALDOA on ferroptosis and ferroptosis-related pathways in GBM cells were further investigated using siRNA/shRNA or CRISPR-Cas9 technology. Results Knockdown of NEDD4L significantly inhibited IKE-induced ferroptosis and reduced lipid peroxidation levels in GBM cells. This study validated NEDD4L as a novel E3 ubiquitin ligase for ALDOA, demonstrating its interaction with ALDOA to mediate monoubiquitination, thereby modulating GBM cell susceptibility to ferroptosis. Additionally, the knockdown of both NEDD4L and ALDOA promoted phosphorylation of 4EBP1 and ACC, and increased the expression of GPX4 protein, which suggested that NEDD4L-mediated ubiquitination of ALDOA might regulate ferroptosis susceptibility in GBM cells through the mTORC1-4EBP1 signalling pathway and lipid metabolism. All data were statistically analyzed using GraphPad Prism 9.0 software. Conclusion NEDD4L has been shown to mediate monoubiquitination of ALDOA, with potential to promote ferroptosis sensitivity by affecting the mTORC1-4EBP1 signaling pathway and lipid metabolism in GBM cells.

Key words: Glioblastoma, Neural precursor cell expressed developmentally down-regulated 4 like, Aldolase A, Ubiquitination, Ferroptosis

CLC Number: 

  • R574
[1] Gritsch S, Batchelor TT, Gonzalez Castro LN. Diagnostic, therapeutic, and prognostic implications of the 2021 World Health Organization classification of tumors of the central nervous system[J]. Cancer, 2022, 128(1): 47-58.
[2] Jiang XJ, Stockwell BR, Conrad M. Ferroptosis: mechanisms, biology and role in disease[J]. Nat Rev Mol Cell Biol, 2021, 22(4): 266-282.
[3] Dixon SJ, Olzmann JA. The cell biology of ferroptosis[J]. Nat Rev Mol Cell Biol, 2024, 25(6): 424-442.
[4] Yee PP, Wei YJ, Kim SY, et al. Neutrophil-induced ferroptosis promotes tumor necrosis in glioblastoma progression[J]. Nat Commun, 2020, 11(1): 5424. doi: 10.1038/s41467-020-19193-y
[5] Liu TQ, Zhu C, Chen X, et al. Ferroptosis, as the most enriched programmed cell death process in glioma, induces immunosuppression and immunotherapy resistance[J]. Neuro Oncol, 2022, 24(7): 1113-1125.
[6] Yan YL, Zhou SJ, Chen X, et al. Suppression of ITPKB degradation by Trim25 confers TMZ resistance in glioblastoma through ROS homeostasis[J]. Signal Transduct Target Ther, 2024, 9(1): 58. doi: 10.1038/s41392-024-01763-x
[7] 洪喆, 毛欣晨, 侯翔云, 等. 脑胶质瘤与铁死亡研究进展 [J]. 肿瘤学杂志, 2024, 30(3): 236-244. HONG Zhe, MAO Xinchen, HOU Xiangyun, et al. Research progress on relationship between brain glioma and ferroptosis [J]. Journal of Chinese Oncology, 2024, 30(3): 236-244.
[8] Minami JK, Morrow D, Bayley NA, et al. CDKN2A deletion remodels lipid metabolism to prime glioblastoma for ferroptosis[J]. Cancer Cell, 2023, 41(6): 1048-1060.e9.
[9] 王文佳, 夏启龙, 张迪, 等. 铁死亡通路泛素化调控的研究进展 [J]. 华中科技大学学报(医学版)2024, 53(4): 528-534. WANG Wenjia, XIA Qilong, ZHANG Di, et al. Research advances in the ubiquitination regulation of ferroptosis pathways [J]. Journal of Huazhong University of Science and Technology(Medical Sciences), 2024, 53(4): 528-534.
[10] Zhou YJ, Zhao Z, Jiang C, et al. LINC01088 prevents ferroptosis in glioblastoma by enhancing SLC7A11 via HLTF/USP7 axis[J]. Clin Transl Med, 2025, 15(3): e70257. doi: 10.1002/ctm2.70257
[11] Li DB, Wang YH, Dong C, et al. CST1 inhibits ferroptosis and promotes gastric cancer metastasis by regulating GPX4 protein stability via OTUB1[J]. Oncogene, 2023, 42(2): 83-98.
[12] Anandhan A, Dodson M, Shakya A, et al. NRF2 controls iron homeostasis and ferroptosis through HERC2 and VAMP8[J]. Sci Adv, 2023, 9(5): eade9585. doi: 10.1126/sciadv.ade9585
[13] Bao ZY, Liu YL, Chen BL, et al. Prokineticin-2 prevents neuronal cell deaths in a model of traumatic brain injury[J]. Nat Commun, 2021, 12(1): 4220. doi: 10.1038/s41467-021-24469-y
[14] Zhou QY, Yu HF, Chen YX, et al. The CRL3KCTD10 ubiquitin ligase-USP18 axis coordinately regulates cystine uptake and ferroptosis by modulating SLC7A11[J]. Proc Natl Acad Sci USA, 2024, 121(28): e2320655121. doi: 10.1073/pnas.2320655121
[15] Yang JJ, Zhou YL, Xie SD, et al. Metformin induces Ferroptosis by inhibiting UFMylation of SLC7A11 in breast cancer[J]. J Exp Clin Cancer Res, 2021, 40(1): 206. doi: 10.1186/s13046-021-02012-7
[16] Liang JJ, Wang N, Yao YH, et al. NEDD4L mediates intestinal epithelial cell ferroptosis to restrict inflammatory bowel diseases and colorectal tumorigenesis[J]. J Clin Invest, 2024, 135(3): e173994. doi: 10.1172/JCI173994
[17] Wang Y, Liu Y, Liu J, et al. NEDD4L-mediated LTF protein degradation limits ferroptosis[J]. Biochem Biophys Res Commun, 2020, 531(4): 581-587.
[18] Chen Z, Wang WL, Hou JH, et al. NEDD4L contributes to ferroptosis and cell growth inhibition in esophageal squamous cell carcinoma by facilitating xCT ubiquitination[J]. Cell Death Discov, 2024, 10: 473. doi: 10.1038/s41420-024-02243-5
[19] Wei YJ, Yee PP, Liu ZJ, et al. NEDD4L-mediated Merlin ubiquitination facilitates Hippo pathway activation[J]. EMBO Rep, 2020, 21(12): e50642. doi: 10.15252/embr.202050642
[20] Valerius AR, Webb LM, Sener U. Novel clinical trials and approaches in the management of glioblastoma[J]. Curr Oncol Rep, 2024, 26(5): 439-465.
[21] 王剑, 周文婧, 薛知, 等. 脑胶质母细胞瘤模型研究概况及类脑模型的研发应用 [J]. 山东大学学报(医学版), 2020, 58(8): 74-80. WANG Jian, ZHOU Wenjing, XUE Zhi, et al. Research overview of glioblastoma models and development application of brain-like models [J]. Journal of Shandong University(Health Sciences), 2020, 58(8): 74-80.
[22] de Souza I, Monteiro LS, Guedes CB, et al. High levels of NRF2 sensitize temozolomide-resistant glioblastoma cells to ferroptosis via ABCC1/MRP1 upregulation[J]. Cell Death Dis, 2022, 13(7): 591. doi: 10.1038/s41419-022-05044-9
[23] Wang YM, Wu XR, Ren Z, et al. Overcoming cancer chemotherapy resistance by the induction of ferroptosis[J]. Drug Resist Updat, 2023, 66: 100916. doi: 10.1016/j.drup.2022.100916
[24] Sehm T, Rauh M, Wiendieck K, et al. Temozolomide toxicity operates in a xCT/SLC7a11 dependent manner and is fostered by ferroptosis[J]. Oncotarget, 2016, 7(46): 74630-74647.
[25] Liu X, Wang W, Nie QC, et al. The role and mechanisms of ubiquitin-proteasome system-mediated ferroptosis in neurological disorders[J]. Neurosci Bull, 2025, 41(4): 691-706.
[26] 陈安静, 张训. 靶向小类泛素化修饰的胶质瘤治疗新策略 [J]. 山东大学学报(医学版), 2020, 58(8): 88-94. CHEN Anjing, ZHANG Xun. Novel therapeutic strategies for glioma targeting SUMOylation [J]. Journal of Shandong University(Health Sciences), 2020, 58(8): 88-94.
[27] He SM, Deng JP, Li G, et al. Down-regulation of Nedd4L is associated with the aggressive progression and worse prognosis of malignant glioma[J]. Jpn J Clin Oncol, 2012, 42(3): 196-201.
[28] Liu J, Kang R, Tang DL. Metabolic checkpoint of ferroptosis resistance[J]. Mol Cell Oncol, 2021, 8(3): 1901558. doi: 10.1080/23723556.2021.1901558
[29] Wu SQ, Mao C, Kondiparthi L, et al. A ferroptosis defense mechanism mediated by glycerol-3-phosphate dehydrogenase 2 in mitochondria[J]. Proc Natl Acad Sci USA, 2022, 119(26): e2121987119. doi: 10.1073/pnas.2121987119
[30] Song XX, Liu J, Kuang FM, et al. PDK4 dictates metabolic resistance to ferroptosis by suppressing pyruvate oxidation and fatty acid synthesis[J]. Cell Rep, 2021, 34(8): 108767. doi: 10.1016/j.celrep.2021.108767
[31] Li MQ, Zhang CS, Feng JW, et al. Aldolase is a sensor for both low and high glucose, linking to AMPK and mTORC1[J]. Cell Res, 2021, 31(4): 478-481.
[32] Luo K, Liu AQ, Wu H, et al. CircKIF4A promotes glioma growth and temozolomide resistance by accelerating glycolysis[J]. Cell Death Dis, 2022, 13(8): 740. doi: 10.1038/s41419-022-05175-z
[33] Ji SR, Zhang B, Liu J, et al. ALDOA functions as an oncogene in the highly metastatic pancreatic cancer[J]. Cancer Lett, 2016, 374(1): 127-135.
[34] Niu Y, Lin ZY, Wan A, et al. Loss-of-function genetic screening identifies aldolase a as an essential driver for liver cancer cell growth under hypoxia[J]. Hepatology, 2021, 74(3): 1461-1479.
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