Skip to main content
SpringerLink
Log in

Decreased Expression of EHD2 Promotes Tumor Metastasis and Indicates Poor Prognosis in Hepatocellular Carcinoma

  • Original Article
  • Published:
Digestive Diseases and Sciences Aims and scope Submit manuscript

Abstract

Background

Metastasis remains the most common cause of lethal outcomes in hepatocellular carcinoma (HCC) after curative resection. Understanding molecular mechanisms that regulate metastasis process is crucial for improving treatment of hepatocellular carcinoma.

Aims

In this article, we examined whether Eps15 homology domain-containing 2 (EHD2) played a critical role in hepatocellular carcinoma metastasis and explored the possible mechanism.

Methods

EHD2 and E-cadherin expression levels in hepatocellular carcinoma patients were examined using Western blotting and immunohistochemistry. The cell migration and invasion were evaluated by wound-healing assay and trans-well assay. Epithelial-mesenchymal transition was analyzed by immunofluorescence, and the vital markers were detected by Western blotting. The correlation of EHD2 and E-cadherin was confirmed by co-immunoprecipitation.

Results

EHD2 expression, along with the epithelial marker E-cadherin, was markedly reduced in tumor tissues than in adjacent noncancerous tissues. Moreover, EHD2 was positively correlated with E-cadherin, histological grade, tumor metastasis, and microvascular invasion. Kaplan–Meier survival analysis showed that hepatocellular carcinoma patients with decreased EHD2 expression had shorter overall survival times than those with higher EHD2 expression. Knockdown of EHD2 induced an increase in cell invasion and changes characteristic of epithelial-mesenchymal transition, while overexpression of EHD2 inhibited these processes.

Conclusions

Molecular data indicated that EHD2 inhibited migration and invasion of hepatocellular carcinoma probably by interacting with E-cadherin and it might be an independent, significant risk factor for survival after curative resection.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

We’re sorry, something doesn't seem to be working properly.

Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

References

  1. Sun H, Gao Y, Lu K, et al. Overexpression of Klotho suppresses liver cancer progression and induces cell apoptosis by negatively regulating wnt/b-catenin signaling pathway. World J Surg Oncol. 2015;13:307.

    Article  PubMed  PubMed Central  Google Scholar 

  2. He L, Zhou X, Qu C, et al. Musashi2 predicts poor prognosis and invasion in hepatocellular carcinoma by driving epithelial-mesenchymal transition. J Cell Mol Med. 2014;18:49–58.

    Article  CAS  PubMed  Google Scholar 

  3. Huang W, Chen Z, Shang X, et al. Sox12, a direct target of FoxQ1, promotes hepatocellular carcinoma metastasis through up-regulating Twist1 and FGFBP1. Hepatology. 2015;61:1920–1933.

    Article  CAS  PubMed  Google Scholar 

  4. Frau M, Biasi F, Feo F, Pascale RM. Prognostic markers and putative therapeutic targets for hepatocellular carcinoma. Mol Aspects Med. 2010;31:179–193.

    Article  CAS  PubMed  Google Scholar 

  5. Maritzen T, Schachtner H, Legler DF. On the move: endocytic trafficking in cell migration. Cell Mol Life Sci. 2015;72:2119–2134.

    Article  CAS  PubMed  Google Scholar 

  6. Peng YF, Shi YH, Ding ZB, et al. Autophagy inhibition suppresses pulmonary metastasis of HCC in mice via impairing anoikis resistance and colonization of HCC cells. Autophagy. 2013;9:2056–2068.

    Article  CAS  PubMed  Google Scholar 

  7. Lu Q, Insinna C, Ott C, et al. Early steps in primary cilium assembly require EHD1/EHD3-dependent ciliary vesicle formation. Nat Cell Biol. 2015;17:228–240.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Naslavsky N, Caplan S. EHD proteins: key conductors of endocytic transport. Trends Cell Biol. 2011;21:122–131.

    Article  CAS  PubMed  Google Scholar 

  9. Marg A, Schoewel V, Timmel T, et al. Sarcolemmal repair is a slow process and includes EHD2. Traffic. 2012;13:1286–1294.

    Article  CAS  PubMed  Google Scholar 

  10. Naslavsky N, Caplan S. C-terminal EH-domain-containing proteins: consensus for a role in endocytic trafficking, EH? J Cell Sci. 2005;118:4093–4101.

    Article  CAS  PubMed  Google Scholar 

  11. Naslavsky N, Rahajeng J, Sharma M, Jovic M, Caplan S. Interactions between EHD proteins and Rab11-FIP2: a role for EHD3 in early endosomal transport. Mol Biol Cell. 2006;17:163–177.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Naslavsky N, McKenzie J, Altan-Bonnet N, Sheff D, Caplan S. EHD3 regulates early-endosome-to-Golgi transport and preserves Golgi morphology. J Cell Sci. 2009;122:389–400.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Barrett T, Troup DB, Wilhite SE, et al. NCBI GEO: archive for functional genomics data sets—10 years on. Nucleic Acids Res. 2011;39:D1005–D1010.

    Article  CAS  PubMed  Google Scholar 

  14. Yang X, Ren H, Yao L, Chen X, He A. Role of EHD2 in migration and invasion of human breast cancer cells. Tumour Biol. 2015;36:3717–3726.

    Article  CAS  PubMed  Google Scholar 

  15. Shi Y, Liu X, Sun Y, et al. Decreased expression and prognostic role of EHD2 in human breast carcinoma: correlation with E-cadherin. J Mol Histol. 2015;46:221–231.

    Article  CAS  PubMed  Google Scholar 

  16. Smith JS, Tachibana I, Pohl U, et al. A transcript map of the chromosome 19q-arm glioma tumor suppressor region. Genomics. 2000;64:44–50.

    Article  CAS  PubMed  Google Scholar 

  17. Li M, Yang X, Zhang J, et al. Effects of EHD2 interference on migration of esophageal squamous cell carcinoma. Med Oncol. 2013;30:396.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Liu J, Ni W, Xiao M, Jiang F, Ni R. Decreased expression and prognostic role of mitogen-activated protein kinase phosphatase 4 in hepatocellular carcinoma. J Gastrointest Surg. 2013;17:756–765.

    Article  PubMed  Google Scholar 

  19. Wan C, Hou S, Ni R, et al. MIF4G domain containing protein regulates cell cycle and hepatic carcinogenesis by antagonizing CDK2-dependent p27 stability. Oncogene. 2015;34:237–245.

    Article  CAS  PubMed  Google Scholar 

  20. Wang Y, Liu F, Mao F, et al. Interaction with cyclin H/cyclin-dependent kinase 7 (CCNH/CDK7) stabilizes C-terminal binding protein 2 (CtBP2) and promotes cancer cell migration. J Biol Chem. 2013;288:9028–9034.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Zhang S, Shi W, Chen Y, et al. Overexpression of SYF2 correlates with enhanced cell growth and poor prognosis in human hepatocellular carcinoma. Mol Cell Biochem. 2015;410:1–9.

    Article  CAS  PubMed  Google Scholar 

  22. Zhang J, Zhu J, Yang L, et al. Interaction with CCNH/CDK7 facilitates CtBP2 promoting esophageal squamous cell carcinoma (ESCC) metastasis via upregulating epithelial-mesenchymal transition (EMT) progression. Tumour Biol. 2015;36:6701–6714.

    Article  CAS  PubMed  Google Scholar 

  23. Schrader J, Gordon-Walker TT, Aucott RL, et al. Matrix stiffness modulates proliferation, chemotherapeutic response, and dormancy in hepatocellular carcinoma cells. Hepatology. 2011;53:1192–1205.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Polyak K, Weinberg RA. Transitions between epithelial and mesenchymal states: acquisition of malignant and stem cell traits. Nat Rev Cancer. 2009;9:265–273.

    Article  CAS  PubMed  Google Scholar 

  25. Li P, Liu X, Dong ZM, Ling ZQ. Epigenetic silencing of HIC1 promotes epithelial-mesenchymal transition and drives progression in esophageal squamous cell carcinoma. Oncotarget. 2015;6:38151–38165.

    PubMed  PubMed Central  Google Scholar 

  26. Fang JH, Zhou HC, Zhang C, et al. A novel vascular pattern promotes metastasis of hepatocellular carcinoma in an epithelial-mesenchymal transition-independent manner. Hepatology. 2015;62:452–465.

    Article  CAS  PubMed  Google Scholar 

  27. Han T, Xiang DM, Sun W, et al. PTPN11/Shp2 overexpression enhances liver cancer progression and predicts poor prognosis of patients. J Hepatol. 2015;63:651–660.

    Article  CAS  PubMed  Google Scholar 

  28. Chen L, Chan TH, Yuan YF, et al. CHD1L promotes hepatocellular carcinoma progression and metastasis in mice and is associated with these processes in human patients. J Clin Invest. 2010;120:1178–1191.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Hu QP, Kuang JY, Yang QK, Bian XW, Yu SC. Beyond a tumor suppressor: soluble E-cadherin promotes the progression of cancer. Int J Cancer. 2015;138:2804–2812.

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by the Natural Science Foundation of China (No. 81401985).

Author information

Authors and Affiliations

  1. Department of Gastroenterology, Affiliated Hospital of Nantong University, 20# Xisi Road, Nantong, 226001, Jiangsu, People’s Republic of China

    Jinxia Liu, Wenkai Ni, Lishuai Qu & Runzhou Ni

  2. Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, 226001, Jiangsu, People’s Republic of China

    Xiaopeng Cui

  3. Grade 14, Clinical Medicine, Medical College, Nantong University, Nantong, 226001, Jiangsu, People’s Republic of China

    Zhipeng Lin, Qingqing Liu & Huiling Zhou

Authors
  1. Jinxia Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wenkai Ni
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lishuai Qu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xiaopeng Cui
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhipeng Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Qingqing Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Huiling Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Runzhou Ni
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Runzhou Ni.

Ethics declarations

Conflict of interest

The authors declare that no conflict of interest exists.

Electronic supplementary material

Below is the link to the electronic supplementary material.

10620_2016_4202_MOESM1_ESM.tif

Fig. S1 a Wound-healing assay. Migration of cells (HepG2, Huh7, Hep3B, SMMC-7721 and SK-Hep1) to the wound was visualized at 0 h and 48 h with an inverted Leica phase-contrast microscope. b The bar chart demonstrated the relative migration distance of cells. The data were mean ± SEM of three independent experiments (*P < 0.05). (TIFF 19788 kb)

10620_2016_4202_MOESM2_ESM.tif

Fig. S2 a Trans-well assay. Invasion of cells (HepG2, Huh7, Hep3B, SMMC-7721 and SK-Hep1) travelled through the micropore membrane was visualized at 48 h. b The bar chart showed the percentage of migrant cells. The data were mean ± SEM of three independent experiments (*, P < 0.05). (TIFF 19190 kb)

10620_2016_4202_MOESM3_ESM.tif

Fig. S3 The bar chart showed the ratio of E-cadherin, Vimentin, N-cadherin, Snail, Slug, Twist and Zeb1 protein to GAPDH by densitometry. The data were mean ± SEM of three independent experiments. (*,#, P < 0.05) (TIFF 13623 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, J., Ni, W., Qu, L. et al. Decreased Expression of EHD2 Promotes Tumor Metastasis and Indicates Poor Prognosis in Hepatocellular Carcinoma. Dig Dis Sci 61, 2554–2567 (2016). https://doi.org/10.1007/s10620-016-4202-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10620-016-4202-6

Keywords

Search

Navigation