Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Phosphorylation enhanced OTUD3 deubiquitination ARID3A promotes the progress of cholangiocarcinoma

Oncogene volume 44, pages 2040–2053 (2025)Cite this article

Subjects

Abstract

Cholangiocarcinoma (CCA) is a highly heterogeneous group of malignant tumors with different molecular etiologies and clinical manifestations. Post-translational modifications (PTM) such as ubiquitination and phosphorylation are widely involved in the progression of CCA. Our aim was to elucidate the effect of the deubiquitinating enzyme OTU domain-containing protein 3 (OTUD3) on the molecular pathogenesis of CAA. OTUD3 and ARID3A exhibit direct binding interactions and cytosolic substructural co-localization in CCA cells.OTUD3 specifically removes the ARID3A proteins K240 and K329 OTUD3 specifically removes the ubiquitinated chains at K240 and K329 of ARID3A protein, thereby enhancing ARID3A stability. OTUD3 promotes CCA proliferation and metastasis in vivo and in vitro by interacting with ARID3A.GSK3β interacts with OTUD3 protein and mediates phosphorylation of the Ser9 site of OTUD3, which enhances the affinity of OTUD3 to ARID3A and further stabilizes ARID3A protein. The expression of OTUD3 and ARID3A in CCA tissues is highly correlated, and the high expression of both proteins is closely related to the poor prognosis of patients. In conclusion, GSK3β phosphorylates OTUD3, enhances its binding ability to ARID3A, and ultimately inhibits the ubiquitination degradation of ARID3A, which promotes the progression of CCA. The GSK3β-OTUD3-ARID3A signaling pathway has been demonstrated for the first time in the progression of CCA.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: OTUD3 interacts with ARID3A.
Fig. 2: OTUD3 stabilizes ARID3A.
Fig. 3: OTUD3 removes the K48-linked ubiquitin chain from K240 and K329 of ARID3A.
Fig. 4: OTUD3 enhances the proliferation, invasion, and metastasis of CCA by stabilizing ARID3A.
Fig. 5: GSK3β phosphorylates OTUD3 to promote the interaction between OTUD3 and ARID3A proteins.
Fig. 6: Pharmacological inhibition of GSK3β suppresses ARID3A stability.
Fig. 7: OTUD3 and ARID3A are correlation with CCA patients survival.

Similar content being viewed by others

Data availability

All data relevant to the study are included in the paper or uploaded as supplementary materials. The data and sources associated with this study are available from the corresponding author upon request.

References

  1. Moris D, Palta M, Kim C, Allen PJ, Morse MA, Lidsky ME. Advances in the treatment of intrahepatic cholangiocarcinoma: An overview of the current and future therapeutic landscape for clinicians. CA Cancer J Clin. 2023;73:198–222.

    Article  PubMed  Google Scholar 

  2. Vithayathil M, Khan SA. Current epidemiology of cholangiocarcinoma in Western countries. J Hepatol. 2022;77:1690–8.

    Article  PubMed  Google Scholar 

  3. Kam AE, Masood A, Shroff RT. Current and emerging therapies for advanced biliary tract cancers. Lancet Gastroenterol Hepatol. 2021;6:956–69.

    Article  PubMed  Google Scholar 

  4. Valle JW, Kelley RK, Nervi B, Oh DY, Zhu AX. Biliary tract cancer. Lancet. 2021;397:428–44.

    Article  CAS  PubMed  Google Scholar 

  5. Li W, Li F, Zhang X, Lin HK, Xu C. Insights into the post-translational modification and its emerging role in shaping the tumor microenvironment. Signal Transduct Target Ther. 2021;6:422.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Chen L, Liu S, Tao Y. Regulating tumor suppressor genes: post-translational modifications. Signal Transduct Target Ther. 2020;5:90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Guo CC, Xu HE, Ma X. ARID3a from the ARID family: structure, role in autoimmune diseases and drug discovery. Acta Pharmacol Sin. 2023;44:2139–50.

  8. Wheat JC, Steidl U. Posttranscriptional Arid3a deregulation in AMKL. Blood. 2022;139:637–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Ratliff ML, Shankar M, Guthridge JM, James JA, Webb CF. TLR engagement induces ARID3a in human blood hematopoietic progenitors and modulates IFNalpha production. Cell Immunol. 2020;357:104201.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Alejo-Valle O, Weigert K, Bhayadia R, Ng M, Issa H, Beyer C, et al. The megakaryocytic transcription factor ARID3A suppresses leukemia pathogenesis. Blood. 2022;139:651–65.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Shen M, Li S, Zhao Y, Liu Y, Liu Z, Huan L, et al. Hepatic ARID3A facilitates liver cancer malignancy by cooperating with CEP131 to regulate an embryonic stem cell-like gene signature. Cell Death Dis. 2022;13:732.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Tang J, Yang L, Li Y, Ning X, Chaulagain A, Wang T, et al. ARID3A promotes the development of colorectal cancer by upregulating AURKA. Carcinogenesis. 2021;42:578–86.

    Article  CAS  PubMed  Google Scholar 

  13. Christine A, Park MK, Song SJ, Song MS. The equilibrium of tumor suppression: DUBs as active regulators of PTEN. Exp Mol Med. 2022;54:1814–21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Du T, Li H, Fan Y, Yuan L, Guo X, Zhu Q, et al. The deubiquitylase OTUD3 stabilizes GRP78 and promotes lung tumorigenesis. Nat Commun. 2019;10:2914.

    Article  PubMed  PubMed Central  Google Scholar 

  15. Wang M, Li Y, Xiao Y, Yang M, Chen J, Jian Y, et al. Nicotine-mediated OTUD3 downregulation inhibits VEGF-C mRNA decay to promote lymphatic metastasis of human esophageal cancer. Nat Commun. 2021;12:7006.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Xie P, Chen Y, Zhang H, Zhou G, Chao Q, Wang J, et al. The deubiquitinase OTUD3 stabilizes ACTN4 to drive growth and metastasis of hepatocellular carcinoma. Aging. 2021;13:19317–38.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Filipcik P, Curry JR, Mace PD. When worlds collide-mechanisms at the interface between phosphorylation and ubiquitination. J Mol Biol. 2017;429:1097–113.

    Article  CAS  PubMed  Google Scholar 

  18. Huang OW, Ma X, Yin J, Flinders J, Maurer T, Kayagaki N, et al. Phosphorylation-dependent activity of the deubiquitinase DUBA. Nat Struct Mol Biol. 2012;19:171–5.

    Article  CAS  PubMed  Google Scholar 

  19. Qin M, Xin Y, Bian Y, Yang X, Xi T, Xiong J. Phosphorylation-induced ubiquitination and degradation of PXR through CDK2-TRIM21 axis. Cells. 2022;11:264.

  20. Feng X, Jia Y, Zhang Y, Ma F, Zhu Y, Hong X, et al. Ubiquitination of UVRAG by SMURF1 promotes autophagosome maturation and inhibits hepatocellular carcinoma growth. Autophagy. 2019;15:1130–49.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Li T, Wang X, Ju E, da Silva SR, Chen L, Zhang X, et al. RNF167 activates mTORC1 and promotes tumorigenesis by targeting CASTOR1 for ubiquitination and degradation. Nat Commun. 2021;12:1055.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Zhang DY, Zhu Y, Wu Q, Ma S, Ma Y, Shen ZC, et al. USP1 promotes cholangiocarcinoma progression by deubiquitinating PARP1 to prevent its proteasomal degradation. Cell Death Dis. 2023;14:669.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Liu WW, Tu JF, Ying XH, Chen ZJ, Wang YB. Postoperative survival of extrahepatic and intrahepatic cholangiocarcinoma after surgery: a population-based cohort. BMJ Open. 2022;12:e049789.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Tran TB, Ethun CG, Pawlik TM, Schmidt C, Beal EW, Fields RC, et al. Actual 5-year survivors after surgical resection of hilar cholangiocarcinoma. Ann Surg Oncol. 2019;26:611–8.

    Article  PubMed  Google Scholar 

  25. Elvevi A, Laffusa A, Scaravaglio M, Rossi RE, Longarini R, Stagno AM, et al. Clinical treatment of cholangiocarcinoma: an updated comprehensive review. Ann Hepatol. 2022;27:100737.

    Article  PubMed  Google Scholar 

  26. Ohaegbulam KC, Koethe Y, Fung A, Mayo SC, Grossberg AJ, Chen EY, et al. The multidisciplinary management of cholangiocarcinoma. Cancer. 2023;129:184–214.

    Article  CAS  PubMed  Google Scholar 

  27. Li Y, Tang J, Li J, Du Y, Bai F, Yang L, et al. ARID3A promotes the chemosensitivity of colon cancer by inhibiting AKR1C3. Cell Biol Int. 2022;46:965–75.

    Article  CAS  PubMed  Google Scholar 

  28. Mao X, Xu J, Xiao M, Liang C, Hua J, Liu J, et al. ARID3A enhances chemoresistance of pancreatic cancer via inhibiting PTEN-induced ferroptosis. Redox Biol. 2024;73:103200.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Pu Q, Lv YR, Dong K, Geng WW, Gao HD. Tumor suppressor OTUD3 induces growth inhibition and apoptosis by directly deubiquitinating and stabilizing p53 in invasive breast carcinoma cells. BMC Cancer. 2020;20:583.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Zhang P, Li C, Li H, Yuan L, Dai H, Peng Z, et al. Ubiquitin ligase CHIP regulates OTUD3 stability and suppresses tumour metastasis in lung cancer. Cell Death Differ. 2020;27:3177–95.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Jin Y, Cheng H, Cao J, Shen W. MicroRNA 32 promotes cell proliferation, migration, and suppresses apoptosis in colon cancer cells by targeting OTU domain containing 3. J Cell Biochem. 2019;120:18629–39.

    Article  CAS  PubMed  Google Scholar 

  32. Zhao M, Yang F, Sang C, Yan C, Wang Z. BGL3 inhibits papillary thyroid carcinoma progression via regulating PTEN stability. J Endocrinol Investig. 2021;44:2165–74.

    Article  CAS  Google Scholar 

  33. Lacoursiere RE, Hadi D, Shaw GS. Acetylation, phosphorylation, ubiquitination (Oh My!): following post-translational modifications on the ubiquitin road. Biomolecules. 2022;12:467.

  34. Murakami T, Felinski EA, Antonetti DA. Occludin phosphorylation and ubiquitination regulate tight junction trafficking and vascular endothelial growth factor-induced permeability. J Biol Chem. 2009;284:21036–46.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Zhao Y, Mudge MC, Soll JM, Rodrigues RB, Byrum AK, Schwarzkopf EA, et al. OTUD4 is a phospho-activated K63 deubiquitinase that regulates MyD88-dependent signaling. Mol Cell. 2018;69:505–16.e5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Zhou N, Qi H, Liu J, Zhang G, Liu J, Liu N, et al. Deubiquitinase OTUD3 regulates metabolism homeostasis in response to nutritional stresses. Cell Metab. 2022;34:1023–41.e8.

    Article  CAS  PubMed  Google Scholar 

  37. Zhou A, Lin K, Zhang S, Chen Y, Zhang N, Xue J, et al. Nuclear GSK3beta promotes tumorigenesis by phosphorylating KDM1A and inducing its deubiquitylation by USP22. Nat Cell Biol. 2016;18:954–66.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Zhang S, Gao W, Tang J, Zhang H, Zhou Y, Liu J, et al. The roles of GSK-3beta in regulation of retinoid signaling and sorafenib treatment response in hepatocellular carcinoma. Theranostics. 2020;10:1230–44.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Tolosa E, Litvan I, Hoglinger GU, Burn D, Lees A, Andres MV, et al. A phase 2 trial of the GSK-3 inhibitor tideglusib in progressive supranuclear palsy. Mov Disord J Mov Disord Soc. 2014;29:470–8.

    Article  CAS  Google Scholar 

  40. Zhang Y, Du T, Liu N, Wang J, Zhang L, Cui CP, et al. Discovery of an OTUD3 inhibitor for the treatment of non-small cell lung cancer. Cell Death Dis. 2023;14:378.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This study was supported by Anhui province health scientific research project (AHWJ2023b015), Anhui Provincial Natural Science Foundation (202308085MH271), Anhui University Natural Science Research Project (2024AH051282), the sixth batch of “special support plan” leading talent projects in Anhui Province.

Author information

Author notes

  1. These authors contributed equally: Deng-Yong Zhang, Yan Zhu, Shuo-Shuo Ma, Chen-yang Xu.

Authors and Affiliations

  1. Department of General Surgery, The First Affiliated Hospital of Bengbu Medical University, Bengbu, Anhui, China

    Deng-Yong Zhang, Shuo-Shuo Ma, Si-hua Liu, Fang-fang Chen & Zheng Lu

  2. College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu, China

    Yan Zhu & Prit Benny Malgulwar

  3. Liver Transplantation Center and HBP Surgery, Sichuan Cancer Hospital & Institute, Sichuan Cancer Center, School of Medicine, University of Electronic Science and Technology of China, Chengdu, China

    Shuo-Shuo Ma, Jin Shang & Xiao-Lun Huang

  4. Medical College, Shaoxing University, Shaoxing, China

    Chen-yang Xu, Hui Wang & Wei-ying Zhao

  5. School of Public Policy & Management, Tsinghua University, Beijing, China

    Zhong-Lin Wang

Authors
  1. Deng-Yong Zhang
    View author publications

    Search author on:PubMed Google Scholar

  2. Yan Zhu
    View author publications

    Search author on:PubMed Google Scholar

  3. Shuo-Shuo Ma
    View author publications

    Search author on:PubMed Google Scholar

  4. Chen-yang Xu
    View author publications

    Search author on:PubMed Google Scholar

  5. Zhong-Lin Wang
    View author publications

    Search author on:PubMed Google Scholar

  6. Hui Wang
    View author publications

    Search author on:PubMed Google Scholar

  7. Si-hua Liu
    View author publications

    Search author on:PubMed Google Scholar

  8. Jin Shang
    View author publications

    Search author on:PubMed Google Scholar

  9. Xiao-Lun Huang
    View author publications

    Search author on:PubMed Google Scholar

  10. Prit Benny Malgulwar
    View author publications

    Search author on:PubMed Google Scholar

  11. Fang-fang Chen
    View author publications

    Search author on:PubMed Google Scholar

  12. Wei-ying Zhao
    View author publications

    Search author on:PubMed Google Scholar

  13. Zheng Lu
    View author publications

    Search author on:PubMed Google Scholar

Contributions

Deng-Yong Zhang, Wei-ying Zhao, Xiao-Lun Huang, and Zheng Lu conceived of and designed the study. Shuoshuo Ma, Yan Zhu, Deng-Yong Zhang, and Chen-yang Xu performed the main experiments, analyzed the data, and drafted the manuscript. Deng-Yong Zhang, Zhonglin Wang, Fangfang Chen, and Hui Wang helped with data analysis. Deng-Yong Zhang, Jin Shang, and Sihua Liu collected samples. Prit Benny Malgulwar assisted with drafting the manuscript. All authors read and agreed to the published version of the paper.

Corresponding authors

Correspondence to Wei-ying Zhao or Zheng Lu.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

This animal study was approved by the Animal Ethics Committee of Bengbu Medical College (No. 2021298). This study was approved by the Bengbu Medical College Ethics Committee (No. 2021230). All methods were performed in accordance with the relevant guidelines and regulations. The informed consent was obtained from all participants.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhang, DY., Zhu, Y., Ma, SS. et al. Phosphorylation enhanced OTUD3 deubiquitination ARID3A promotes the progress of cholangiocarcinoma. Oncogene 44, 2040–2053 (2025). https://doi.org/10.1038/s41388-025-03376-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue date:

  • DOI: https://doi.org/10.1038/s41388-025-03376-2

Search

Advanced search

Quick links