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TRIM21-mediated ubiquitination of SIX2 attenuates breast cancer stemness via LGSN suppression

Oncogene (2025)Cite this article

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Abstract

Breast cancer stem cells (BCSCs) are pivotal drivers of breast tumor initiation, metastasis, and therapy resistance. Our previous studies identified the transcription factor SIX2 as a key regulator in maintaining breast cancer stemness. Here, we demonstrate that TRIM21, as an E3 ubiquitin ligase downregulated in breast cancer tissues, binds to SIX2 via its PRY-SPRY domain and catalyzes K48-type ubiquitination at lysine residues K82, K89, and K97. This modification promotes the degradation of SIX2 via the ubiquitin-proteasome pathway, consequently attenuating the stemness and metastatic potential of breast cancer cells. Furthermore, SIX2 transcriptionally activates LGSN expression through direct binding to its promoter region, thereby promoting the stemness and metastatic capabilities of breast cancer cells. Clinically, elevated expression of both SIX2 and LGSN correlates with poor prognosis in breast cancer patients. These results establish that TRIM21-mediated degradation of SIX2 suppresses LGSN expression, ultimately inhibiting the stemness and metastatic abilities of breast cancer cells, underscoring the critical regulatory role of the TRIM21-SIX2-LGSN axis in breast cancer progression.

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Fig. 1: E3 ubiquitin ligase TRIM21 directly interacts with SIX2.
Fig. 2: TRIM21 degrades SIX2 via the ubiquitin-proteasome pathway.
Fig. 3: TRIM21-mediated ubiquitination impairs SIX2 -induced enhancement of breast cancer stemness.
Fig. 4: TRIM21-mediated ubiquitination degradation impairs SIX2-induced metastasis of breast cancer cells.
Fig. 5: TRIM21 induces SIX2 ubiquitination through its RING structural domain C16A and thus impairs the stemness and metastasis of breast cancer cells.
Fig. 6: SIX2 knockdown-mediated attenuation of tumorigenic and metastatic ability can be rescued by TRIM21 knockdown.
Fig. 7: LGSN is identified as a direct downstream effector of SIX2.
Fig. 8: SIX2 promotes breast cancer cell stemness and metastatic capacity dependent on LGSN.
Fig. 9

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All data are presented within the article and supplementary online data.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant No. 82204432, 82473955, 82173842), the Science and Technology Research Project of Henan Province (No. 252102311191, No. 242102310257), the Fundamental Research Funds for the Central Universities (2632025TD04), and the Priority Academic Program Development (PAPD) of Jiangsu Higher Education Institutions.

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  1. These authors contributed equally: Haitao Chen, Yi Zhou, Yunnan Zhang.

Authors and Affiliations

  1. School of Life Science and Technology, China Pharmaceutical University, Nanjing, China

    Haitao Chen, Yi Zhou & Lufeng Zheng

  2. Department of Pharmacy, The Affiliated Cancer Hospital of Zhengzhou University & Henan Cancer Hospital, Zhengzhou, PR China

    Yunnan Zhang, Yannan Fan & Qianqian Guo

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Lufeng Zheng and Qianqian Guo designed the research. Haitao Chen, Yi Zhou analyzed the data. Haitao Chen, Yi Zhou, Yunnan Zhang, Yannan Fan performed the research. Haitao Chen and Yi Zhou wrote the paper. Lufeng Zheng, Qianqian Guo reviewed this paper. All authors read and approved the final manuscript.

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All methods were performed in accordance with the relevant guidelines and regulations. All animal care and use procedures were conducted in accordance with the Principles of Animal Care established by China Pharmaceutical University and the Guide for the Care and Use of Laboratory Animals. The study protocol was approved by the Ethics Committee of China Pharmaceutical University (Approval Number: 2025-01-011). All methods were performed in compliance with relevant guidelines and regulations. Informed consent was obtained from all participants involved in the study.

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Chen, H., Zhou, Y., Zhang, Y. et al. TRIM21-mediated ubiquitination of SIX2 attenuates breast cancer stemness via LGSN suppression. Oncogene (2025). https://doi.org/10.1038/s41388-025-03572-0

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