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Cellular and Molecular Biology

NDRG1 facilitates self-renewal of liver cancer stem cells by preventing EpCAM ubiquitination

British Journal of Cancer volume 129, pages 237–248 (2023)Cite this article

Subjects

Abstract

Background

Portal vein tumour thrombus (PVTT) is the main pathway of HCC intrahepatic metastasis and is responsible for the poor prognosis of patients with HCC. However, the molecular mechanisms underlying PVTT vascular metastases have not been fully elucidated.

Methods

NDRG1 expression was assessed by immunohistochemistry and immunoblotting in clinical specimens obtained from curative surgery. The functional relevance of NDRG1 was evaluated using sphere formation and animal models of tumorigenicity and metastasis. The relationship between NDRG1 and EpCAM was explored using molecular biological techniques.

Results

NDRG1 protein was upregulated in HCC samples compared to non-tumorous tissues. Furthermore, NDRG1 expression was enhanced in the PVTT samples. Our functional study showed that NDRG1 was required for the self-renewal of tumour-initiating/cancer stem cells (CSCs). In addition, NDRG1 knockdown inhibited the proliferation and migration of PVTT-1 cells in vitro and in vivo. NDRG1 was found to stabilise the functional tumour-initiating cell marker EpCAM through protein–protein interactions and inhibition of EpCAM ubiquitination.

Conclusion

Our findings suggest that NDRG1 enhances CSCs expansion, PVTT formation and growth capability through the regulation of EpCAM stability. NDRG1 may be a promising target for the treatment of patients with HCC and PVTT.

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Fig. 1: NDRG1 expression is upregulated in HCC and indicates the unfavourable prognosis of patients with HCC.
Fig. 2: NDRG1 expression is upregulated in PVTT samples compared to primary cancer and para-cancerous tissues.
Fig. 3: NDRG1 promotes tumour proliferation and carcinogenesis in HCC.
Fig. 4: NDRG1 enhances tumour-initiating capacity in HCC tumours.
Fig. 5: EpCAM is a functional target of NDRG1 in HCC.
Fig. 6: NDRG1 interacts with and increases EpCAM protein stability.

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Data availability

All results presented in the article are available upon request from the corresponding author.

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Acknowledgements

We would like to thank the Prof. Hongyang Wang (Shanghai Eastern Hepatobiliary Surgery Hospital) for providing technical support.

Funding

This work was supported by National Natural Science Foundation of China (81502581, 82172693, 81872508) and the China Postdoctoral Science Foundation (2022M711911).

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Author notes

  1. These authors contributed equally: Qian Cheng, Shanglei Ning.

Authors and Affiliations

  1. Department of Hepatobiliary Surgery, Beijing Key Laboratory of HCC and Liver Cirrhosis, Peking University People’s Hospital, 100044, Beijing, China

    Qian Cheng & Jiye Zhu

  2. Key Laboratory of Infection and Immunity, Institute of Biophysics, Chinese Academy of Sciences, 100101, Beijing, China

    Qian Cheng, Lei Zhu, Changlu Zhang & Shaodong Jiang

  3. Department of General Surgery, Qilu Hospital of Shandong University, 250012, Shandong, China

    Shanglei Ning

  4. Key Laboratory of RNA Biology, Center for Big Data Research in Health, Institute of Biophysics, Chinese Academy of Sciences, 100101, Beijing, China

    Yajing Hao

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Contributions

QC and JZ designed the work and provided material support. QC and SN acquired the main data. LZ performed several molecular biology experiments. CZ and SJ performed several cell experiments. YH provided bioinformatics analysis. QC, LZ and SN drafted the manuscript. JZ revised the manuscript.

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Correspondence to Qian Cheng or Jiye Zhu.

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The authors declare no competing interests.

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The experiments involved in clinical specimens were performed in accordance with the Declaration of Helsinki and were approved by the Research Ethics Committee of Peking University People’s Hospital. Informed consent was obtained from all participants. In addition, the animal studies were approved by the Institutional Animal Welfare and Ethics Committee of Peking University People’s Hospital.

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Cheng, Q., Ning, S., Zhu, L. et al. NDRG1 facilitates self-renewal of liver cancer stem cells by preventing EpCAM ubiquitination. Br J Cancer 129, 237–248 (2023). https://doi.org/10.1038/s41416-023-02278-y

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