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Digestive Diseases and Sciences

, Volume 61, Issue 9, pp 2554–2567 | Cite as

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

  • Jinxia Liu
  • Wenkai Ni
  • Lishuai Qu
  • Xiaopeng Cui
  • Zhipeng Lin
  • Qingqing Liu
  • Huiling Zhou
  • Runzhou NiEmail author
Original Article

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.

Keywords

EHD2 E-cadherin Metastasis Hepatocellular carcinoma 

Notes

Acknowledgments

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

Compliance with ethical standards

Conflict of interest

The authors declare that no conflict of interest exists.

Supplementary material

10620_2016_4202_MOESM1_ESM.tif (19.3 mb)
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 (18.7 mb)
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 (13.3 mb)
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)

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Jinxia Liu
    • 1
  • Wenkai Ni
    • 1
  • Lishuai Qu
    • 1
  • Xiaopeng Cui
    • 2
  • Zhipeng Lin
    • 3
  • Qingqing Liu
    • 3
  • Huiling Zhou
    • 3
  • Runzhou Ni
    • 1
    Email author
  1. 1.Department of GastroenterologyAffiliated Hospital of Nantong UniversityNantongPeople’s Republic of China
  2. 2.Department of General SurgeryAffiliated Hospital of Nantong UniversityNantongPeople’s Republic of China
  3. 3.Grade 14, Clinical Medicine, Medical CollegeNantong UniversityNantongPeople’s Republic of China

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