Skip to main content

Thank you for visiting 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.

Cellular and Molecular Biology

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



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.


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.


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.


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.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Prices vary by article type



Prices may be subject to local taxes which are calculated during checkout

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.

Data availability

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


  1. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68:394–424.

    Article  PubMed  Google Scholar 

  2. Loh JJ, Li TW, Zhou L, Wong TL, Liu X, Ma VW, et al. FSTL1 secreted by activated fibroblasts promotes hepatocellular carcinoma metastasis and stemness. Cancer Res. 2021;81:5692–705.

    Article  CAS  PubMed  Google Scholar 

  3. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. CA A Cancer J Clin. 2019;69:7–34.

    Article  Google Scholar 

  4. Yang P, Li QJ, Feng Y, Zhang Y, Markowitz GJ, Ning S, et al. TGF-β-miR-34a-CCL22 signaling-induced treg cell recruitment promotes venous metastases of HBV-positive hepatocellular carcinoma. Cancer Cell. 2012;22:291–303.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Fan X, Li Y, Yi X, Chen G, Jin S, et al. Epigenome-wide DNA methylation profiling of portal vein tumor thrombosis (PVTT) tissues in hepatocellular carcinoma patients. Neoplasia. 2020;22:630–43.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Zheng H, Pomyen Y, Hernandez MO, Li C, Livak F, Tang W, et al. Single-cell analysis reveals cancer stem cell heterogeneity in hepatocellular carcinoma. Hepatology. 2018;68:127–40.

    Article  PubMed  Google Scholar 

  7. Yamashita T, Wang XW. Cancer stem cells in the development of liver cancer. J Clin Invest. 2013;123:1911–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Ling S, Shan Q, Zhan Q, Ye Q, Liu P, Xu S, et al. USP22 promotes hypoxia-induced hepatocellular carcinoma stemness by a HIF1α/USP22 positive feedback loop upon TP53 inactivation. Gut. 2020;69:1322–34.

    Article  CAS  PubMed  Google Scholar 

  9. Yamashita T, Ji J, Budhu A, Forgues M, Yang W, Wang HY, et al. EpCAM-positive hepatocellular carcinoma cells are tumor-initiating cells with stem/progenitor cell features. Gastroenterology. 2009;136:1012–24.

    Article  CAS  PubMed  Google Scholar 

  10. Tang KH, Ma S, Lee TK, Chan YP, Kwan PS, Tong CM, et al. CD133(+) liver tumor-initiating cells promote tumor angiogenesis, growth, and self-renewal through neurotensin/interleukin-8/CXCL1 signaling. Hepatology. 2012;55:807–20.

    Article  CAS  PubMed  Google Scholar 

  11. Asai R, Tsuchiya H, Amisaki M, Makimoto K, Takenaga A, Sakabe T, et al. CD44 standard isoform is involved in maintenance of cancer stem cells of a hepatocellular carcinoma cell line. Cancer Med. 2019;8:773–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Mani SK, Zhang H, Diab A, Pascuzzi PE, Lefrançois L, Fares N, et al. EpCAM‑regulated intramembrane proteolysis induces a cancer stem cell‑like gene signature in hepatitis B virus‑infected hepatocytes. J Hepatol. 2016;65:888–98.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Münz M, Kieu C, Mack B, Schmitt B, Zeidler R, Gires O. The carcinoma-associated antigen EpCAM upregulates c-myc and induces cell proliferation. Oncogene. 2004;23:5748–58.

    Article  PubMed  Google Scholar 

  14. Nio K, Yamashita T, Okada H, Kondo M, Hayashi T, Hara Y, et al. Defeating EpCAM(+) liver cancer stem cells by targeting chromatin remodeling enzyme CHD4 in human hepatocellular carcinoma. J Hepatol. 2015;63:1164–72.

    Article  CAS  PubMed  Google Scholar 

  15. Bae DH, Jansson PJ, Huang ML, Kovacevic Z, Kalinowski D, Lee CS, et al. The role of NDRG1 in the pathology and potential treatment of human cancers. J Clin Pathol. 2013;66:911–7.

    Article  CAS  PubMed  Google Scholar 

  16. Zhang P, Tchou-Wong KM, Costa M. Egr-1 mediates hypoxia-inducible transcription of the NDRG1 gene through an overlapping Egr-1/Sp1 binding site in the promoter. Cancer Res. 2007;67:9125–33.

    Article  CAS  PubMed  Google Scholar 

  17. Lane DJ, Saletta F, Suryo Rahmanto Y, Kovacevic Z, Richardson DR. N-myc downstream regulated 1 (NDRG1) is regulated by eukaryotic initiation factor 3a (eIF3a) during cellular stress caused by iron depletion. PLoS ONE. 2013;8:e57273.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Merlot AM, Porter GM, Sahni S, Lim EG, Peres P, Richardson DR. The metastasis suppressor, NDRG1, differentially modulates the endoplasmic reticulum stress response. Biochim Biophys Acta Mol Basis Dis. 2019;1865:2094–110.

    Article  CAS  PubMed  Google Scholar 

  19. Sevinsky CJ, Khan F, Kokabee L, Darehshouri A, Maddipati KR, Conklin DS. NDRG1 regulates neutral lipid metabolism in breast cancer cells. Breast Cancer Res. 2018;20:55.

    Article  PubMed  PubMed Central  Google Scholar 

  20. Villodre ES, Hu X, Eckhardt BL, Larson R, Huo L, Yoon EC, et al. NDRG1 in aggressive breast cancer progression and brain metastasis. J Natl Cancer Inst. 2022;114:579–91.

    Article  PubMed  Google Scholar 

  21. Ito H, Watari K, Shibata T, Miyamoto T, Murakami Y, Nakahara Y, et al. Bidirectional regulation between NDRG1 and GSK3b controls tumor growth and is targeted by differentiation inducing factor-1 in glioblastoma. Cancer Res. 2020;80:234–48.

    Article  CAS  PubMed  Google Scholar 

  22. Mi L, Zhu F, Yang X, Lu J, Zheng Y, Zhao Q, et al. The metastatic suppressor NDRG1 inhibits EMT, migration and invasion through interaction and promotion of caveolin-1 ubiquitylation in human colorectal cancer cells. Oncogene. 2017;36:4323–35.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Maruyama Y, Ono M, Kawahara A, Yokoyama T, Basaki Y, Kage M, et al. Tumor growth suppression in pancreatic cancer by a putative metastasis suppressor gene Cap43/NDRG1/Drg-1 through modulation of angiogenesis. Cancer Res. 2006;66:6233–42.

    Article  CAS  PubMed  Google Scholar 

  24. Bandyopadhyay S, Pai SK, Hirota S, Hosobe S, Takano Y, Saito K, et al. Role of the putative tumor metastasis suppressor gene Drg-1 in breast cancer progression. Oncogene. 2004;23:5675–81.

    Article  CAS  PubMed  Google Scholar 

  25. Ai R, Sun Y, Guo Z, Wei W, Zhou L, Liu F, et al. NDRG1 overexpression promotes the progression of esophageal squamous cell carcinoma through modulating Wnt signaling pathway. Cancer Biol Ther. 2016;17:943–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Lu WJ, Chua MS, So SK. Suppressing N-Myc downstream regulated gene 1 reactivates senescence signaling and inhibits tumor growth in hepatocellular carcinoma. Carcinogenesis. 2014;35:915–22.

    Article  CAS  PubMed  Google Scholar 

  27. Berghoff AS, Liao Y, Karreman MA, Ilhan-Mutlu A, Gunkel K, Sprick MR, et al. Identification and characterization of cancer cells that initiate metastases to the brain and other organs. Mol Cancer Res. 2021;19:688–701.

    Article  CAS  PubMed  Google Scholar 

  28. Wang T, Hu HS, Feng YX, Shi J, Li N, Guo WX, et al. Characterisation of a novel cell line (CSQT-2) with high metastatic activity derived from portal vein tumour thrombus of hepatocellular carcinoma. Br J Cancer. 2010;102:1618–26.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Tang Y, Liu S, Li N, Guo W, Shi J, Yu H, et al. 14-3-3ζ promotes hepatocellular carcinoma venous metastasis by modulating hypoxia-inducible factor-1α. Oncotarget. 2016;7:15854–67.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Hu Y, Smyth GK. ELDA: Extreme limiting dilution analysis for comparing depleted and enriched populations in stem cell and other assays. J Immunol Methods. 2016;347:70–8.

    Article  Google Scholar 

  31. Wang Y, Zhu P, Luo J, Wang J, Liu Z, Wu W, et al. LncRNA HAND2-AS1 promotes liver cancer stem cell self-renewal via BMP signaling. EMBO J. 2019;38:e101110.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Cheng J, Xie HY, Xu X, Wu J, Wei X, Su R, et al. NDRG1 as a biomarker for metastasis, recurrence and of poor prognosis in hepatocellular carcinoma. Cancer Lett. 2011;310:35–45.

    Article  CAS  PubMed  Google Scholar 

  33. Dang H, Chen L, Tang P, Cai X, Zhang W, Zhang R, et al. LINC01419 promotes cell proliferation and metastasis in hepatocellular carcinoma by enhancing NDRG1 promoter activity. Cell Oncol. 2020;43:931–47.

    Article  CAS  Google Scholar 

  34. Akiba J, Ogasawara S, Kawahara A, Nishida N, Sanada S, Moriya F, et al. N-myc downstream regulated gene 1 (NDRG1)/Cap43 enhances portal vein invasion and intrahepatic metastasis in human hepatocellular carcinoma. Oncol Rep. 2008;20:1329–35.

    PubMed  Google Scholar 

  35. Chang X, Xu X, Xue X, Ma J, Li Z, Deng P, et al. N-myc downstream-regulated gene 1 promotes tumor inflammatory angiogenesis through JNK activation and autocrine loop of interleukin-1α by human gastric cancer cells. J Biol Chem. 2013;288:25025–37.

    Article  Google Scholar 

  36. Wang Y, Zhou Y, Tao F, Chai S, Xu X, Yang Y, et al. N-myc downstream regulated gene 1(NDRG1) promotes the stem-like properties of lung cancer cells through stabilized c-Myc. Cancer Lett. 2017;401:53–62.

    Article  CAS  PubMed  Google Scholar 

  37. Zhao T, Meng Y, Wang Y, Wang W. NDRG1 regulates osteosarcoma cells via mediating the mitochondrial function and CSCs differentiation. J Orthop Surg Res. 2021;16:364.

    Article  PubMed  PubMed Central  Google Scholar 

  38. Wangpu X, Yang X, Zhao J, Lu J, Guan S, Lu J, et al. The metastasis suppressor, NDRG1, inhibits “stemness” of colorectal cancer via down-regulation of nuclear beta-catenin and CD44. Oncotarget. 2015;6:33893–911.

    Article  PubMed  PubMed Central  Google Scholar 

Download references


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


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

Author information

Authors and Affiliations



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.

Corresponding authors

Correspondence to Qian Cheng or Jiye Zhu.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

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.

Consent for publication

Not applicable.

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

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


Quick links