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.

Identification of a novel Calpain-2-SRC feed-back loop as necessity for β-Catenin accumulation and signaling activation in hepatocellular carcinoma

Abstract

Rapid progression is the major cause of the poor prognosis of hepatocellular carcinoma (HCC); however, the underlying mechanism remained unclear. Here, we found Calpain-2 (CAPN2), a well-established protease that accelerates tumor progression in several malignancies, is overexpressed in HCC and acts as an independent predictor for poor outcomes. Furthermore, CAPN2 promoted the proliferation and invasion of HCC, and showed a positive correlation with the levels of invasion-related markers. Mechanistically, a novel CAPN2-SRC positive regulatory loop was identified upstream of β-catenin to prevent its ubiquitination and degradation, and subsequently promoted HCC progression: CAPN2 could proteolyze PTP1B to form a truncation of approximately 42 kDa with increased phosphatase activity, resulting in reduced SRC Y530 phosphorylation and increased SRC kinase activity; meanwhile, CAPN2 itself was a bone fide substrate of SRC that was primarily phosphorylated at Y625 by SRC and exhibited increased proteolysis activity upon phosphorylation. Interestingly, the CAPN2-SRC loop could not only restrain most of cytoplasmic β-catenin degradation by inhibiting GSK3β pathway, but also prevented TRIM33-induced nuclear β-catenin degradation even in β-catenin-mutant cells. Present study identified a CAPN2-SRC positive loop responsible for intracellular β-catenin accumulation and signaling activation, and targeting CAPN2 protease activity might be a promising approach for preventing HCC progression.

Your institute does not have access to this article

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Fig. 1: CAPN2 was preferentially expressed in hepatocellular carcinoma (HCC) and predicted poor outcomes.
Fig. 2: CAPN2 promoted HCC progression in vitro and in vivo.
Fig. 3: CAPN2 enhanced β-catenin signaling to promote HCC progression.
Fig. 4: CAPN2 prevented β-catenin from ubiquitination-mediated degradation.
Fig. 5: CAPN2 primarily enhanced β-catenin signaling via activating SRC.
Fig. 6: The CAPN2/SRC regulatory axis enhanced β-catenin signaling directly by phosphorylating β-catenin and indirectly through PI3K/AKT.
Fig. 7: The CAPN2/SRC axis prevented β-catenin degradation by both GSK3β-dependent and -independent pathways.
Fig. 8: SRC transactivated CAPN2 primarily by phosphorylating it at Y625, thus forming a CAPN2/SRC positive regulatory loop.

Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

  1. Zhu XD, Sun HC. Emerging agents and regimens for hepatocellular carcinoma. J Hematol Oncol. 2019;12:110.

    Article  Google Scholar 

  2. Finn RS, Zhu AX. Evolution of systemic therapy for hepatocellular carcinoma. Hepatology. 2021;73:150–7.

    Article  Google Scholar 

  3. Lee TK, Guan XY, Ma S. Cancer stem cells in hepatocellular carcinoma - from origin to clinical implications. Nat Rev Gastroenterol Hepatol. 2022;19:26–44.

  4. Ahn JC, Teng PC, Chen PJ, Posadas E, Tseng HS, Lu S, et al. Detection of circulating tumor cells and their implications as a biomarker for diagnosis, prognostication, and therapeutic monitoring in hepatocellular carcinoma. Hepatology. 2021;73:422–36.

    Article  Google Scholar 

  5. Kong FH, Ye QF, Miao XY, Liu X, Huang SQ, Xiong L, et al. Current status of sorafenib nanoparticle delivery systems in the treatment of hepatocellular carcinoma. Theranostics. 2021;11:5464–90.

    CAS  Article  Google Scholar 

  6. Ye QH, Zhu WW, Zhang JB, Qin Y, Lu M, Lin GL, et al. GOLM1 modulates EGFR/RTK cell-surface recycling to drive hepatocellular carcinoma metastasis. Cancer Cell. 2016;30:444–58.

    CAS  Article  Google Scholar 

  7. Baudry M, Bi X. Calpain-1 and Calpain-2: the Yin and Yang of synaptic plasticity and neurodegeneration. Trends Neurosci. 2016;39:235–45.

    CAS  Article  Google Scholar 

  8. Xie XQ, Wang MJ, Li Y, Lei LP, Wang N, Lv ZY, et al. miR-124 intensified oxaliplatin-based chemotherapy by targeting CAPN2 in colorectal cancer. Mol Ther Oncolytics. 2020;17:320–31.

    CAS  Article  Google Scholar 

  9. Xu F, Gu J, Lu C, Mao W, Wang L, Zhu QL, et al. Calpain-2 enhances non-small cell lung cancer progression and chemoresistance to paclitaxel via EGFR-pAKT pathway. Int J Biol Sci. 2019;15:127–37.

    CAS  Article  Google Scholar 

  10. Storr SJ, Lee KW, Woolston CM, Safuan S, Green AR, Macmillan RD, et al. Calpain system protein expression in basal-like and triple-negative invasive breast cancer. Ann Oncol. 2012;23:2289–96.

    CAS  Article  Google Scholar 

  11. Lee WJ, Shin CH, Ji H, Jeong SD, Park MS, Won HH, et al. hnRNPK-regulated LINC00263 promotes malignant phenotypes through miR-147a/CAPN2. Cell Death Dis. 2021;12:290.

    CAS  Article  Google Scholar 

  12. Zhang Y, Zhu X, Qiao X, Gu X, Xue JQ, Han YS, et al. LIPH promotes metastasis by enriching stem-like cells in triple-negative breast cancer. J Cell Mol Med. 2020;24:9125–34.

    CAS  Article  Google Scholar 

  13. Shen C, Yu Y, Li H, Liu MQ, Shen HL, Yang PY. Global profiling of proteolytically modified proteins in human metastatic hepatocellular carcinoma cell lines reveals CAPN2 centered network. Proteomics. 2012;12:1917–27.

    CAS  Article  Google Scholar 

  14. Perugorria MJ, Olaizola P, Labiano I, Esparza-Baquer A, Marzioni M, Marin JJG, et al. Wnt-β-catenin signalling in liver development, health and disease. Nat Rev Gastroenterol Hepatol. 2019;16:121–36.

    CAS  Article  Google Scholar 

  15. Liu Y, Chen H, Zheng P, Zheng YX, Luo Q, Xie GH, et al. ICG-001 suppresses growth of gastric cancer cells and reduces chemoresistance of cancer stem cell-like population. J Exp Clin Cancer Res. 2017;36:125.

    Article  Google Scholar 

  16. Liu Q, He L, Li S, Li FY, Deng GZ, Huang XJ, et al. HOMER3 facilitates growth factor-mediated β-Catenin tyrosine phosphorylation and activation to promote metastasis in triple negative breast cancer. J Hematol Oncol. 2021;14:6.

    CAS  Article  Google Scholar 

  17. Moro L, Simoneschi D, Kurz E, Arbini AA, Jang SW, Guaragnella N, et al. Epigenetic silencing of the ubiquitin ligase subunit FBXL7 impairs c-SRC degradation and promotes epithelial-to-mesenchymal transition and metastasis. Nat Cell Biol. 2020;22:1130–42.

    CAS  Article  Google Scholar 

  18. Leung HW, Leung CON, Lau EY, Chung KPS, Mok EH, Lei MML, et al. EPHB2 activates β-catenin to enhance cancer stem cell properties and drive sorafenib resistance in hepatocellular carcinoma. Cancer Res. 2021;81:3229–40.

    CAS  Article  Google Scholar 

  19. Pleiman CM, Hertz WM, Cambier JC. Activation of phosphatidylinositol-3’ kinase by Src-family kinase SH3 binding to the p85 subunit. Science. 1994;263:1609–12.

    CAS  Article  Google Scholar 

  20. Ngan E, Stoletov K, Smith HW, Common J, Muller WJ, Lewis JD, et al. LPP is a Src substrate required for invadopodia formation and efficient breast cancer lung metastasis. Nat Commun. 2017;8:15059.

    Article  Google Scholar 

  21. Liu L, Wang T, Yang X, Xu CX, Liao ZH, Wang XD, et al. MTNR1B loss promotes chordoma recurrence by abrogating melatonin-mediated β-catenin signaling repression. J Pineal Res. 2019;67:e12588.

    Article  Google Scholar 

  22. Lin J, Song T, Li C, Mao W. GSK-3β in DNA repair, apoptosis, and resistance of chemotherapy, radiotherapy of cancer. Biochim Biophys Acta Mol Cell Res. 2020;1867:118659.

    CAS  Article  Google Scholar 

  23. Song Y, Liu Y, Pan S, Xie S, Wang ZW, Zhu X. Role of the COP1 protein in cancer development and therapy. Semin Cancer Biol. 2020;67:43–52.

    CAS  Article  Google Scholar 

  24. Lyle CL, Belghasem M, Chitalia VC. c-Cbl: an important regulator and a target in angiogenesis and tumorigenesis. Cells. 2019;8:498.

  25. Xue J, Chen Y, Wu Y, Wang ZY, Zhou AD, Zhang SC, et al. Tumour suppressor TRIM33 targets nuclear β-catenin degradation. Nat Commun. 2015;6:6156.

    CAS  Article  Google Scholar 

  26. Cortesio CL, Chan KT, Perrin BJ, Burton NO, Zhang S, Zhang ZY, et al. Calpain 2 and PTP1B function in a novel pathway with Src to regulate invadopodia dynamics and breast cancer cell invasion. J Cell Biol. 2008;180:957–71.

    CAS  Article  Google Scholar 

  27. Blom N, Sicheritz-Pontén T, Gupta R, Gammeltoft S, Brunak S. Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence. Proteomics. 2004;4:1633–49.

    CAS  Article  Google Scholar 

  28. Wang C, Xu H, Lin S, Deng WK, Zhou JQ, Zhang Y, et al. GPS 5.0: an update on the prediction of kinase-specific phosphorylation sites in proteins. Genomics Proteom Bioinforma. 2020;18:72–80.

    CAS  Article  Google Scholar 

  29. Gao Q, Zhu H, Dong L, Shi WW, Chen R, Song ZJ, et al. Integrated proteogenomic characterization of HBV-related hepatocellular carcinoma. Cell. 2019;179:561–77.

    CAS  Article  Google Scholar 

  30. Caruso S, Calatayud AL, Pilet J, Le Bella T, Rekik S, Imbeaud S, et al. Analysis of liver cancer cell lines identifies agents with likely efficacy against hepatocellular carcinoma and markers of response. Gastroenterology. 2019;157:760–76.

    CAS  Article  Google Scholar 

  31. Chitalia VC, Foy RL, Bachschmid MM, Zeng LL, Panchenko MV, Zhou MI, et al. Jade-1 inhibits Wnt signalling by ubiquitylating beta-catenin and mediates Wnt pathway inhibition by pVHL. Nat Cell Biol. 2008;10:1208–16.

    CAS  Article  Google Scholar 

  32. Shao H, Chou J, Baty CJ, Burke NA, Watkins SC, Stolz DB, et al. Spatial localization of m-calpain to the plasma membrane by phosphoinositide biphosphate binding during epidermal growth factor receptor-mediated activation. Mol Cell Biol. 2006;26:5481–96.

    CAS  Article  Google Scholar 

  33. Tai WT, Chen YL, Chu PY, Chen LJ, Hung MH, Shiau CW, et al. Protein tyrosine phosphatase 1B dephosphorylates PITX1 and regulates p120RasGAP in hepatocellular carcinoma. Hepatology. 2016;63:1528–43.

    CAS  Article  Google Scholar 

  34. Bruix J, Sherman M. Management of hepatocellular carcinoma: an update. Hepatology. 2011;53:1020–2.

    Article  Google Scholar 

  35. Jiang Y, Sun A, Zhao Y, Ying WT, Sun HC, Yang XR, et al. Proteomics identifies new therapeutic targets of early-stage hepatocellular carcinoma. Nature. 2019;567:257–61.

    CAS  Article  Google Scholar 

  36. Sun YF, Xu Y, Yang XR, Guo W, Zhang X, Qiu SJ, et al. Circulating stem cell-like epithelial cell adhesion molecule-positive tumor cells indicate poor prognosis of hepatocellular carcinoma after curative resection. Hepatology. 2013;57:1458–68.

    CAS  Article  Google Scholar 

  37. Dong ZR, Sun D, Yang YF, Zhou W, Wu R, Wang XW, et al. TMPRSS4 drives angiogenesis in hepatocellular carcinoma by promoting HB-EGF expression and proteolytic cleavage. Hepatology. 2020;72:923–39.

    CAS  Article  Google Scholar 

  38. Hu B, Xu Y, Li YC, Huang JF, Cheng JW, Guo W, et al. CD13 promotes hepatocellular carcinogenesis and sorafenib resistance by activating HDAC5-LSD1-NF-κB oncogenic signaling. Clin Transl Med. 2020;10:e233.

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank all the patient enrolled for their agreement for using their tissue samples. Moreover, we thank James P. Mahaffey, PhD, from Liwen Bianji (Edanz) (www.liwenbianji.cn/), for editing the English text of a draft of this manuscript.

Funding

LR-Q was sponsored by National Natural Science Foundation of China (81772774, 82072876); Program of scientific and technological innovation action plan, Shanghai Municipal Commission of science and technology (20XD1434200, 20Y11903500). GL was sponsored by National Natural Science Foundation of China (81772808). RN was sponsored by the Shanghai International Science and Technology Collaboration Program (18410721900), and the National Natural Science Foundation of China (81472672). MX-L was sponsored by Shanghai Sailing Program (20YF1407900); National Natural Science Foundation of China (82002618). WY-C was sponsored by National Natural Science Foundation of China (81800190). ZH was sponsored by Shanghai Natural Science Foundation, Shanghai Municipal Commission of science and technology (19ZR1410300).

Author information

Authors and Affiliations

Authors

Contributions

RQL, LG, NR, and XLM contributed to the study design. XLM, KYZ, YDC, YT, HZ, YCW, and WGT contributed to the acquisition, analysis, and interpretation of the data. XLM, SHX, NR, LG, and RQL helped to draft the manuscript. NR, LG, and RQL critically revised the manuscript. All authors read and approve the final manuscript.

Corresponding authors

Correspondence to Ning Ren, Lin Guo or Ren-Quan Lu.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

This study was approved by the Shanghai Cancer Center Research Ethics Committee and Zhongshan Hospital Research Ethics Committee, and all individuals provided informed consent for inclusion of their tissue in this study. Moreover, animal study was approved by the Animal Experimentation Ethics Committee of Shanghai Cancer Center, Fudan University.

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

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Ma, XL., Zhu, KY., Chen, YD. et al. Identification of a novel Calpain-2-SRC feed-back loop as necessity for β-Catenin accumulation and signaling activation in hepatocellular carcinoma. Oncogene 41, 3554–3569 (2022). https://doi.org/10.1038/s41388-022-02367-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-022-02367-x

Search

Quick links