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Epithelial membrane protein 2: a novel biomarker for circulating tumor cell recovery in breast cancer

  • Q. Chen
  • L. Yao
  • D. Burner
  • B. Minev
  • L. Lu
  • M. Wang
  • W. Ma
Research Article

Abstract

Purpose

EpCAM is a common marker used in the detection of circulating tumor cells (CTC). Disseminated cancer cells display the characteristics of epithelial-to-mesenchymal transition events. The purpose of this study was to assess the potential of epithelial membrane protein 2 (EMP2) as a novel biomarker for CTC retrieval in breast cancer.

Methods

MCF7 and MDA-MB-231 cells were stained with either anti-EpCAM or anti-EMP2 mAbs, respectively, followed by flow cytometric assay to measure their expression levels. PBMCs isolated from healthy donors were used for breast cancer cell spiking. CD45-depleted PBMCs from breast cancer patients’ blood were used for CTC capturing. Immunomagnetic separation was used to enrich breast cancer cells. Cytospin centrifugation was performed to concentrate the captured cells, followed by immunofluorescence staining with anti-CD45 mAb, anti-pan cytokeratin mAb and DAPI. Fluorescent images were taken using a confocal microscope for CTC counts.

Result

MDA-MB-231 cells had 2.56 times higher EMP2 expression than MCF7 cells, and EMP2 had a significantly higher capture efficiency than EpCAM for MCF7 cells. Furthermore, anti-EMP2 was capable of capturing MCF7 cells that escaped in the flow-through of anti-EpCAM. Likewise, EMP2 had a significantly higher capture efficiency on MDA-MB-231 cells when compared to MCF7 cells. Most importantly, EMP2 biomarker was successfully used for CTC capture in patients with primary breast cancer.

Conclusions

EMP2 is superior to EpCAM for capturing both MCF7 and MDA-MB-231 cells. Additionally, EMP2 is a novel biomarker and capable of capturing breast cancer cells in patient blood samples.

Keywords

Epithelial membrane protein 2 Circulating tumor cells Breast cancer Biomarker Capture 

Notes

Acknowledgements

The authors would like to thank Dr. Shengwen Shao at the Institute of Microbiology and Immunology, Huzhou University School of Medicine for his professional help on cell-staining and images taking.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical statement

This study was reviewed and approved by the ethics committee of the institutional review board (IRB, HU2014-04), the First Hospital Affiliated to Huzhou University School of Medicine. All procedures performed in this study involving human participants were in accordance with the ethical standards of the institutional and/or national research committee.

Informed consent

Informed consent forms were obtained from all individual participants who were included in the study.

Supplementary material

12094_2018_1941_MOESM1_ESM.docx (114 kb)
Supplementary material 1 (DOCX 114 kb)

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Copyright information

© Federación de Sociedades Españolas de Oncología (FESEO) 2018

Authors and Affiliations

  1. 1.Key Laboratory for Translational Medicine and the Division of Breast SurgeryThe First Affiliated Hospital of Huzhou University School of MedicineHuzhouChina
  2. 2.Department of Medicine and Moores Cancer CenterUniversity of California San DiegoLa JollaUSA
  3. 3.Calidi BiotherapeuticsSan DiegoUSA
  4. 4.Department of Chronic Disease Epidemiology, Yale School of Public Health, Yale School of MedicineYale Cancer CenterNew HavenUSA
  5. 5.Division of Gastrointestinal SurgeryThe First Affiliated Hospital and the Department of Clinical MedicineJiaxing University School of MedicineJiaxingChina

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