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:

SPRTN is involved in hepatocellular carcinoma development through the ER stress response

Cancer Gene Therapy volume 31, pages 376–386 (2024)Cite this article

Subjects

A Correction to this article was published on 10 April 2024

This article has been updated

Abstract

Endoplasmic reticulum (ER) stress, prompted by the accumulation of misfolded or unfolded proteins, triggers the activation of the unfolded protein response (UPR) pathway to restore ER homeostasis. This stress response is implicated in the development of hepatocellular carcinoma (HCC). A biallelic mutation in SPRTN is currently the only known single-gene mutation implicated in the early onset of HCC. However, the exact mechanism linking SPRTN mutations to HCC remains unclear. In our study, we analyzed SPRTN and UPR in 21 human HCC tissue samples using RT-qPCR, immunoblot, and immunohistochemistry. We found alterations in the expression levels of SPRTN and UPR-related genes and proteins in HCC samples. The impact of SPRTN on the ER stress response was assessed in SPRTN-depleted HepG2 cells through RNA sequencing, pull-down assay, comet assay, and mitotic index calculation. We demonstrated that SPRTN interacts with the UPR sensor GRP78. Furthermore, we observed a decrease in SPRTN levels during ER stress, and increased sensitivity to ER stress in SPRTN-depleted cells. These findings suggest an essential role for SPRTN in the ER stress response and provide new insights into HCC pathogenesis. This newly discovered function of SPRTN could significantly enhance our understanding and treatment of HCC.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Fig. 1: SPRTN is involved in the ER stress response.
Fig. 2: Gene expression and protein level analysis under ER stress.
Fig. 3: SPRTN and ER stress response factors are altered in HCC.
Fig. 4: ER stress is a key element of HCC and SPRTN is essential for preventing DNA damage during ER stress.
Fig. 5: RNA degradation and protein aggregation in ER stress response are influenced by SPRTN.

Similar content being viewed by others

Data availability

The RNA sequencing dataset generated and analyzed during the current study is available in the Gene Expression Omnibus (GEO, NCBI) using GEO Series accession number GSE237978. The human hepatocellular carcinoma dataset used for the comparison during the current study is available in the Cancer Genome Atlas (TCGA) repository: https://www.cancer.gov/tcga.

Change history

References

  1. McGlynn KA, Petrick JL, El-Serag HB. Epidemiology of hepatocellular carcinoma. Hepatology. 2021;73:4–13.

    Article  CAS  PubMed  Google Scholar 

  2. Xie JL, Jarosz DF. Mutations, protein homeostasis, and epigenetic control of genome integrity. DNA Repair. 2018;71:23–32. https://doi.org/10.1016/j.dnarep.2018.08.004.

    Article  CAS  PubMed  Google Scholar 

  3. Hetz C, Papa FR. The unfolded protein response and cell fate control. Mol Cell. 2018;69:169–81.

    Article  CAS  PubMed  Google Scholar 

  4. Marciniak SJ. Endoplasmic reticulum stress: a key player in human disease. FEBS J. 2019;286:228–31.

    Article  CAS  PubMed  Google Scholar 

  5. Liu X, Green RM. Endoplasmic reticulum stress and liver diseases. Liver Res. 2019;3:55–64.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Wei J, Fang D. Endoplasmic reticulum stress signaling and the pathogenesis of hepatocarcinoma. Int J Mol Sci. 2021;22:1–15.

    Google Scholar 

  7. Pavlović N, Heindryckx F. Targeting ER stress in the hepatic tumor microenvironment. FEBS J. 2021;289:7163–76.

    Article  PubMed  Google Scholar 

  8. Rauschert N, Brändlein S, Holzinger E, Hensel F, Müller-Hermelink HK, Vollmers HP. A new tumor-specific variant of GRP78 as target for antibody-based therapy. Lab Investig. 2008;88:375–86.

    Article  CAS  PubMed  Google Scholar 

  9. Pavlović N, Heindryckx F. Exploring the role of endoplasmic reticulum stress in hepatocellular carcinoma through mining of the human protein atlas. Biology (Basel). 2021;10:640

    PubMed  Google Scholar 

  10. Cook KL, Soto-Pantoja DR, Clarke PAG, Cruz MI, Zwart A, Wärri A, et al. Endoplasmic reticulum stress protein GRP78 modulates lipid metabolism to control drug sensitivity and antitumor immunity in breast cancer. Cancer Res. 2016;76:5657–70.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Ron D, Walter P. Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol. 2007;8:519–29.

    Article  CAS  PubMed  Google Scholar 

  12. Lu Y, Cederbaum AI. CYP2E1 and oxidative liver injury by alcohol. Free Radic Biol Med. 2008;44:723–38.

    Article  CAS  PubMed  Google Scholar 

  13. Ren X, Ji Y, Jiang X, Qi X. Downregulation of CYP2A6 and CYP2C8 in tumor tissues is linked to worse overall survival and recurrence-free survival from hepatocellular carcinoma. Biomed Res Int. 2018;2018:1–9.

    Article  CAS  Google Scholar 

  14. Sampieri L, Di Giusto P, Alvarez C. CREB3 transcription factors: ER-golgi stress transducers as hubs for cellular homeostasis. Front Cell Dev Biol. 2019;7:1–13.

    Article  Google Scholar 

  15. Lopez-Mosqueda J, Maddi K, Prgomet S, Kalayil S, Marinovic-Terzic I, Terzic J, et al. SPRTN is a mammalian DNA-binding metalloprotease that resolves DNA-protein crosslinks. Elife. 2016;5:1–19.

    Article  Google Scholar 

  16. Vaz B, Popovic M, Newman JA, Fielden J, Aitkenhead H, Halder S, et al. Metalloprotease SPRTN/DVC1 orchestrates replication-coupled DNA-protein crosslink repair. Mol Cell. 2016;64:704–19. https://doi.org/10.1016/j.molcel.2016.09.032.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Perry M, Ghosal G. Mechanisms and regulation of DNA-protein crosslink repair during DNA replication by SPRTN protease. Front Mol Biosci. 2022;9:1–18.

    Article  Google Scholar 

  18. Ruijs MWG, van Andel RNJ, Oshima J, Madan K, Nieuwint AWM, Aalfs CM. Atypical progeroid syndrome: An unknown helicase gene defect? Am J Med Genet. 2003;116A:295–9.

    Article  CAS  PubMed  Google Scholar 

  19. Lessel D, Vaz B, Halder S, Lockhart PJ, Marinovic-Terzic I, Lopez-Mosqueda J, et al. Mutations in SPRTN cause early onset hepatocellular carcinoma, genomic instability and progeroid features. Nat Genet. 2014;46:1239–44.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Glumac M, Polović M, Batel A, Gelemanović A, Maček B, Velić A, et al. SPRTN-dependent DPC degradation precedes repair of damaged DNA: a proof of concept revealed by the STAR assay. Nucleic Acids Res. 2023;51:1–18.

    Article  Google Scholar 

  21. Oughtred R, Rust J, Chang C, Breitkreutz BJ, Stark C, Willems A, et al. The BioGRID database: a comprehensive biomedical resource of curated protein, genetic, and chemical interactions. Protein Sci. 2021;30:187–200.

    Article  CAS  PubMed  Google Scholar 

  22. Gillespie M, Jassal B, Stephan R, Milacic M, Rothfels K, Senff-Ribeiro A, et al. The reactome pathway knowledgebase 2022. Nucleic Acids Res. 2022;50:D687–92.

    Article  CAS  PubMed  Google Scholar 

  23. Perry M, Biegert M, Kollala SS, Mallard H, Su G, Kodavati M, et al. USP11 mediates repair of DNA–protein cross-links by deubiquitinating SPRTN metalloprotease. J Biol Chem. 2021;296:100396. https://doi.org/10.1016/j.jbc.2021.100396.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Leliveld SR, Bader V, Hendriks P, Prikulis I, Sajnani G, Requena JR, et al. Insolubility of disrupted-in-schizophrenia 1 disrupts oligomer-dependent interactions with nuclear distribution element 1 and is associated with sporadic mental disease. J Neurosci. 2008;28:3839–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Zaharija B, Odorčić M, Hart A, Samardžija B, Marreiros R, Prikulis I, et al. TRIOBP-1 protein aggregation exists in both major depressive disorder and schizophrenia, and can occur through two distinct regions of the protein. Int J Mol Sci. 2022;23:1–19.

    Article  Google Scholar 

  26. Wu J, Chen S, Liu H, Zhang Z, Ni Z, Chen J, et al. Tunicamycin specifically aggravates ER stress and overcomes chemoresistance in multidrug-resistant gastric cancer cells by inhibiting N-glycosylation. J Exp Clin Cancer Res. 2018;37:1–12.

    Article  Google Scholar 

  27. Sehgal P, Szalai P, Olesen C, Praetorius HA, Nissen P, Christensen SB, et al. Inhibition of the sarco/endoplasmic reticulum (ER) Ca2-ATPase by thapsigargin analogs induces cell death via ER Ca2 depletion and the unfolded protein response. J Biol Chem. 2017;292:19656–73.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Casas C. GRP78 at the centre of the stage in cancer and neuroprotection. Front Neurosci. 2017;11:1–15.

    Article  Google Scholar 

  29. Yamaguchi H, Wang HG. CHOP is involved in endoplasmic reticulum stress-induced apoptosis by enhancing DR5 expression in human carcinoma cells. J Biol Chem. 2004;279:45495–502. https://doi.org/10.1074/jbc.M406933200.

    Article  CAS  PubMed  Google Scholar 

  30. Moslehi A, Hamidi-Zad Z. Role of SREBPs in liver diseases: a mini-review. J Clin Transl Hepatol. 2018;6:332–8.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Gramantieri L, Chieco P, Giovannini C, Lacchini M, Treré D, Grazi GL, et al. GADD45-α expression in cirrhosis and hepatocellular carcinoma: relationship with DNA repair and proliferation. Hum Pathol. 2005;36:1154–62.

    Article  CAS  PubMed  Google Scholar 

  32. Xun R, Lu H, Wang X. Identification of CDC25C as a potential biomarker in hepatocellular carcinoma using bioinformatics analysis. Technol Cancer Res Treat. 2020;19:1–10.

    Article  Google Scholar 

  33. Hong J, Hu K, Yuan Y, Sang Y, Bu Q, Chen G, et al. CHK1 targets spleen tyrosine kinase (L) for proteolysis in hepatocellular carcinoma. J Clin Invest. 2012;122:2165–75.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Mah LJ, El-Osta A, Karagiannis TC. γh2AX: A sensitive molecular marker of DNA damage and repair. Leukemia. 2010;24:679–86. https://doi.org/10.1038/leu.2010.6.

    Article  CAS  PubMed  Google Scholar 

  35. Butler P, Pascheto I, Lizzi M, Onge RS, Lanner C, Guo B, et al. RNA disruption is a widespread phenomenon associated with stress ‑ induced cell death in tumour cells. Sci Rep. 2023;13:1–16. https://doi.org/10.1038/s41598-023-28635-8.

    Article  CAS  Google Scholar 

  36. Schroeder A, Mueller O, Stocker S, Salowsky R, Leiber M, Gassmann M, et al. The RIN: An RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol. 2006;7:1–14.

    Article  Google Scholar 

  37. Maskey RS, Flatten KS, Sieben CJ, Peterson KL, Baker DJ, Nam HJ, et al. Spartan deficiency causes accumulation of Topoisomerase 1 cleavage complexes and tumorigenesis. Nucleic Acids Res. 2017;45:4564–76.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Vandewynckel YP, Laukens D, Bogaerts E, Paridaens A, Van den Bussche A, Verhelst X, et al. Modulation of the unfolded protein response impedes tumor cell adaptation to proteotoxic stress: a PERK for hepatocellular carcinoma therapy. Hepatol Int. 2015;9:93–104.

    Article  PubMed  Google Scholar 

  39. Jaud M, Philippe C, Di Bella D, Tang W, Pyronnet S, Laurell H, et al. Translational regulations in response to endoplasmic reticulum stress in cancers. Cells. 2020;9:1–28.

    Article  Google Scholar 

  40. Yamamori T, Meike S, Nagane M, Yasui H, Inanami O. ER stress suppresses DNA double-strand break repair and sensitizes tumor cells to ionizing radiation by stimulating proteasomal degradation of Rad51. FEBS Lett. 2013;587:3348–53. https://doi.org/10.1016/j.febslet.2013.08.030.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Prof. Ivan Dikic for his valuable advice and support.

Funding

Funding was provided by the Croatian Science Foundation HRZZ (IP-2016-06-3097) and the University of Split School of Medicine. The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.

Author information

Author notes

  1. These authors contributed equally: Anja Batel, Mirjana Polović, Mateo Glumac.

Authors and Affiliations

  1. Laboratory for Cancer Research, University of Split School of Medicine, Šoltanska 2, 21000, Split, Croatia

    Anja Batel, Mirjana Polović, Mateo Glumac, Bernarda Lozić & Ivana Marinović Terzić

  2. Department of Abdominal Surgery, Merkur Clinical Hospital, Zajčeva 19, 10000, Zagreb, Croatia

    Oliver Šuman & Stipislav Jadrijević

  3. Laboratory for Human Genetics, University Hospital Split, Spinčićeva 1, 21000, Split, Croatia

    Bernarda Lozić & Marija Petrović

  4. Faculty of Biotechnology & Drug Development, University of Rijeka, Radmile Matejčić 2, 51000, Rijeka, Croatia

    Bobana Samardžija & Nicholas J. Bradshaw

  5. Selvita, Prilaz baruna Filipovića 29, 10000, Zagreb, Croatia

    Karlo Skube

  6. Department of Physiology, Faculty of Medicine, University of Helsinki, Haartmaninkatu 8, 00290, Helsinki, Finland

    Vinko Palada

  7. Omics solutions, trg 101. Brigade HV 1, 10000, Zagreb, Croatia

    Mislav Acman

Authors
  1. Anja Batel
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mirjana Polović
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mateo Glumac
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Oliver Šuman
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Stipislav Jadrijević
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Bernarda Lozić
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Marija Petrović
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Bobana Samardžija
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Nicholas J. Bradshaw
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Karlo Skube
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Vinko Palada
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Mislav Acman
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Ivana Marinović Terzić
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors contributed to the study conceptualization and design, material preparation, data collection, and analysis. The original draft of the manuscript was written by Anja Batel, Mirjana Polović, Mateo Glumac, and Ivana Marinović Terzić. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Ivana Marinović Terzić.

Ethics declarations

Competing interests

The authors declare no competing interests.

Compliance with ethical standards

The study protocol was approved by the Medical Ethics Committee of the University of Split, School of Medicine (003-08/20-03/0005) and the Clinical Hospital Merkur, Zagreb (0311-7732-3) in accordance with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Consent to participate

Informed written consent for the collection of liver specimens was obtained from each individual participant included in the study. No individual person’s data in any form, including individual details, images or videos was used in the publication.

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

Batel, A., Polović, M., Glumac, M. et al. SPRTN is involved in hepatocellular carcinoma development through the ER stress response. Cancer Gene Ther 31, 376–386 (2024). https://doi.org/10.1038/s41417-023-00708-w

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41417-023-00708-w

Search

Advanced search

Quick links