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


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.

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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.


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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) (, for editing the English text of a draft of this manuscript.


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).

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Authors and Affiliations



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.

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

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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.

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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).

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