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Ephrin-A2 promotes prostate cancer metastasis by enhancing angiogenesis and promoting EMT

  • Original Article – Cancer Research
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

Background

Ephrin-A2, a member of the Eph receptor subgroup, is used in diagnosing and determining the prognosis of prostate cancer. However, the role of ephrin-A2 in prostate cancer is remains elusive.

Methods

We established stable clones overexpressing or silencing ephrin-A2 from prostate cancer cells. Then, CCK-8 was used in analyzing the proliferation ability of cells. CD31 staining was used in evaluating angiogenesis. Migration and invasion assay were conducted in vivo and in vitro. The expression of EMT-related markers was evaluated in prostate cancer cells through Western blotting.

Results

We revealed that the ectopic expression of ephrin-A2 in prostate cancer cells facilitated cell migration and invasion in vitro and promoted tumor metastasis and angiogenesis in vivo and that the silencing of ephrin-A2 completely reversed this effect. Although ephrin-A2 did not affect tumor cell proliferation in vitro, ephrin-A2 significantly promoted primary tumor growth in vivo. Furthermore, to determine the biological function of ephrin-A2, we assayed the expression of EMT-related markers in stable-established cell lines. Results showed that the overexpression of ephrin-A2 in prostate cancer cells down-regulated the expression of epithelial markers (ZO-1, E-cadherin, and claudin-1) and up-regulated the expression of mesenchymal markers (N-cadherin, β-catenin, vimentin, Slug, and Snail), but the knocking out of ephrin-A2 opposed the effects on the expression of EMT markers.

Conclusions

These findings indicate that ephrin-A2 promotes prostate cancer metastasis by enhancing angiogenesis and promoting EMT and may be a potentially therapeutic target in metastatic prostate cancer.

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Data availability statement

The raw data used and analyzed in the current study are available from the corresponding author upon a reasonable request.

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Funding

This study was supported by the grant from the Postdoctoral Science Foundation of China (No.2020M670076ZX), the National Natural Science Foundation of China (No. 81702061), the Natural Science Foundation of Jiangsu Province (No. BK20160233) and the Jiangsu Provincial Medical Youth Talent (No. QNRC2016781).

Author information

Author notes

  1. Yao Zhao and Chenchen Cai contributed equally to this work.

Authors and Affiliations

  1. Department of Clinical Laboratory, The Second Affiliated Hospital of Soochow University, Suzhou, 215004, China

    Yao Zhao

  2. Medical Technology School of Xuzhou Medical University, Xuzhou, 221004, China

    Yao Zhao, Chenchen Cai, Miaomiao Zhang, Lubing Shi, Jiwei Wang, Ping Ma & Shibao Li

  3. Xuzhou Central Hospital, The Affiliated Xuzhou Hospital of Medical College of Southeast University, Xuzhou, 221002, China

    Chenchen Cai

  4. Department of Laboratory Medicine, Affiliated Hospital of Xuzhou Medical University, 99 West Huaihai Road, Xuzhou, 221002, China

    Haoliang Zhang, Ping Ma & Shibao Li

Authors
  1. Yao Zhao
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  2. Chenchen Cai
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  3. Miaomiao Zhang
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  4. Lubing Shi
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  7. Ping Ma
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Contributions

SBL, YZ and CCC contributed to the idea, drafting and editing of the manuscript. LUS and JWW completed statistical analysis. MMZ and HLZ contributed to literature search and feeding animals and data analysis. PM and SBL revised the manuscript with critical reviews and comments and all the authors approved the final version.

Corresponding authors

Correspondence to Ping Ma or Shibao Li.

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Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Ethical statement

The animals in this study were obtained in accordance with the ethical standards of the ethics committee of the affiliated Hospital of Xuzhou Medical University.

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Supplementary Information

Below is the link to the electronic supplementary material.

432_2021_3618_MOESM1_ESM.tif

Supplementary file1 Supporting Fig. 1 Ephrin-A2 promotes tumor angiogenesis. VEGF was evaluated in tumor tissues of tumor-bearing mice (DU145 group) through immunohistochemical staining. The criterion for a positive immune reaction for VEGF is a dark brown cytoplasmic precipitate. Scale bar, 100 µm. (TIF 1420 KB)

432_2021_3618_MOESM2_ESM.tif

Supplementary file2 Supporting Fig. 2 Effects of EphrinA2 on migration of PC-3 cells in vitro. Wound areas of overexpression ephrin-A2 PC-3 cells. The cells were cultured in regular medium and wound healing abilities were quantified by measuring the average gap area between 0 and 24 h with ImageJ. (TIF 179 KB)

432_2021_3618_MOESM3_ESM.tif

Supplementary file3 Supporting Fig. 3 Effects of EphrinA2 on migration of LNCaP cells in vitro. Wound areas of knockdown ephrin-A2 LNCaP cells. The cells were cultured in regular medium and wound healing abilities were quantified by measuring the average gap area between 0 and 24 h with ImageJ. (TIF 187 KB)

432_2021_3618_MOESM4_ESM.tif

Supplementary file4 Supporting Fig. 4 Effects of EphrinA2 on migration of DU145 cells in vitro. Wound areas of knockdown ephrin-A2 DU145 cells. The cells were cultured in regular medium and wound healing abilities were quantified by measuring the average gap area between 0 and 24 h with ImageJ. (TIF 159 KB)

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Zhao, Y., Cai, C., Zhang, M. et al. Ephrin-A2 promotes prostate cancer metastasis by enhancing angiogenesis and promoting EMT. J Cancer Res Clin Oncol 147, 2013–2023 (2021). https://doi.org/10.1007/s00432-021-03618-2

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  • DOI: https://doi.org/10.1007/s00432-021-03618-2

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