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A20 interacts with mTORC2 to inhibit the mTORC2/Akt/Rac1 signaling axis in hepatocellular carcinoma cells

Cancer Gene Therapy volume 30, pages 424–436 (2023)Cite this article

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Abstract

A20 acts as a tumor suppressor in hepatocellular carcinoma, especially inhibiting metastasis of the malignant cells. However, the mechanisms whereby A20 plays the inhibitory roles are not understood completely. Rac1 signaling is essential for cell migration in hepatocellular carcinoma metastasis. Nevertheless, it is not known whether and how A20 inhibits Rac1 signaling to suppress the migration of hepatocellular carcinoma cell. Thereby, we analyzed the relationship between A20 and Rac1 activation, as well as the activity of Akt and mTORC2, two signaling components upstream of Rac1, using gain and loss of function experiments. We found that the overexpression of A20 repressed, while the knockdown or knockout of A20 promoted, the activation of Rac1, Akt and mTORC2 in hepatocellular carcinoma cells. Moreover, the inhibitory effect of A20 on the mTORC2/Akt/Rac1 signaling axis was due to the interaction between A20 and mTORC2 complex. The binding of A20 to mTORC2 was mediated by the ZnF7 domain of A20 and M1 ubiquitin chain in the mTORC2 complex. Furthermore, A20 inhibited metastasis of hepatocellular carcinoma cells via restraining mTORC2 in a hepatocellular carcinoma xenograft mouse model. These findings revealed the relationship between A20 and mTORC2, and explained the molecular mechanisms of A20 in inhibition of hepatocellular carcinoma metastasis.

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Fig. 1: A20 inhibits HCC cell migration by suppressing Rac1 activation in vitro and in vivo.
Fig. 2: A20 inhibits Rac1 activity via suppressing Akt-activation in HCC cells.
Fig. 3: A20 inhibits the activity of mTORC2.
Fig. 4: A20 binds to mTORC2 complex.
Fig. 5: A20 interacts with mTORC2 via the ZnF7 domain of A20 and the M1 linear ubiquitin in the mTORC2.
Fig. 6: A20 inhibits migration of HCC cells via restraining mTORC2 activity in vitro and in vivo.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

This work was supported by the National Nature Science Foundation of China [grant numbers 81672806 and 81972685] and the Key Research and Development Program of Shandong Province [grant number 2017GSF218066].

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Authors and Affiliations

  1. Department of Immunology and Shandong Key Laboratory of Infection and Immunity, School of Basic Medical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, 250012, China

    Xinyu Wang, Yanlei Dong, Jing Yi, Jianing Wang, Xiaoyan Wang, Lining Zhang & Yongyu Shi

  2. Laboratory of Cellular and Molecular Medicine, School of Basic Medical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China

    Ying Xiao

  3. Department of Clinical Laboratory, Shandong Second Provincial General Hospital, Jinan, China

    Zhida Wang

  4. Department of Pathogen Biology, School of Basic Medical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, China

    Huaiyu Zhou

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Contributions

XW designed and carried out most of the experiments, and wrote the original draft. YX and ZW contributed to cell culture. YD and JY performed in vivo experiments. JW and XW contributed to data analysis. HZ and LZ contributed to interpretation of the results and writing-reviewing. YS conceived the study, supervised the research and wrote the paper.

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Correspondence to Yongyu Shi.

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All procedures for animal experiments were approved by the Animal Care and Utilization Committee of Shandong University and performed in a manner compliant with all relevant ethical regulations regarding animal research.

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Wang, X., Xiao, Y., Dong, Y. et al. A20 interacts with mTORC2 to inhibit the mTORC2/Akt/Rac1 signaling axis in hepatocellular carcinoma cells. Cancer Gene Ther 30, 424–436 (2023). https://doi.org/10.1038/s41417-022-00562-2

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