Phosphorylation of EphA2 receptor and vasculogenic mimicry is an indicator of poor prognosis in invasive carcinoma of the breast

  • Debarpan Mitra
  • Sayantan Bhattacharyya
  • Neyaz Alam
  • Sagar Sen
  • Saunak Mitra
  • Syamsundar Mandal
  • Shivani Vignesh
  • Biswanath Majumder
  • Nabendu MurmuEmail author
Preclinical study



The occurrence of vasculogenic mimicry (VM) and EphA2-mediated tumour progression are associated with poor prognosis in various solid tumours. Here, we aimed to investigate the prognostic implications of VM and its association with phosphorylated EphA2 receptor in invasive carcinoma of the breast.


The patients were stratified based on CD-31/PAS dual staining and subsequently the expression status of phospho-EphA2 (S897), FAK, phospho-ERK1/2 and Laminin 5Ƴ2 was analysed by immunohistochemistry. Survival of patients was correlated within the stratified cohort.


The pathologically defined VM phenotype and phospho-EphA2 (S897) expression status were significantly associated with lower disease-free survival (DFS) and overall survival (OS). Both the features were also found to be significantly associated with higher nodal status, poor Nottingham Prognostic Index (NPI) and were more prevalent in the triple-negative breast cancer (TNBC) group. Incidentally, there were no significant association between age of the patient, grade and size of the tumour with VM and phospho-EphA2 (S897). The effector molecules of phospho-EphA2 (S897) viz., Focal Adhesion Kinase (FAK), phospho-ERK1/2 and Laminin 5Ƴ2 were significantly upregulated in the VM-positive cohort. Survival analysis revealed that the VM and phospho-EphA2 (S897) dual-positive cohort had poorest DFS [mean time = 48.313 (39.992–56.633) months] and OS [mean time = 56.692 (49.055–64.328) months]. Individually, VM-positive [Hazard Ratio (HR) 6.005; 95% confidence interval (CI) 2.002–18.018; P = 0.001 for DFS and HR 11.654; 95% CI 3.195–42.508; P < 0.0001 for OS] and phospho-EphA2 (S897)-positive (HR 4.342; 95% CI 1.717–10.983; P = 0.002 for DFS and HR 5.853; 95% CI 1.663–20.602; P = 0.006 for OS) expression proved to be independent indicators of prognosis.


This study evaluated tumour dependency on oncogenic EphA2 receptor regulation and VM in invasive carcinoma of the breast and their prognostic significance. Significant correlations between VM, phospho-EphA2 and several clinicopathologic parameters of breast cancer were found. Subsequently, the occurrence of VM or phospho-EphA2 expression proved to be major contributors for poor prognosis in patients with breast cancer but their simultaneous expression failed to be an independent risk factor.


Breast cancer Vasculogenic mimicry EphA2 Prognosis Overall survival 



We wish to thank Dr. Jayanta Chakrabarti, Director, CNCI, for his immense support throughout the project.

Author contributions

DM and NM conceptualised and designed the study. DM and SB performed IHC experiments and collected patient data. DM and SSM analysed the data. DM, SM, SV, BM and NM interpreted the data. NA and SS provided clinical insights and interpreted clinical data and did follow-up of patients. DM, SB, BM and NM wrote the manuscript.


This project was supported by Chittaranjan National Cancer Institute, Kolkata (Intramural fund, Sanction No. A-4.482/2017/271).

Compliance with ethical standards

Conflict of interest

Authors have no conflict of interest to declare.

Ethical approval

This study was approved by the Institutional ethics committee at Chittaranjan National Cancer Institute, Kolkata (IEC Ref: A-4.311/NM/26/11/2018). All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Informed consent

Written informed consent was not required as this was a retrospective study and patient information was de-identified prior to analysis.

Supplementary material

10549_2019_5482_MOESM1_ESM.tif (888 kb)
Supplementary material 1 (TIFF 888 kb)
10549_2019_5482_MOESM2_ESM.doc (33 kb)
Supplementary material 2 (DOC 33 kb)
10549_2019_5482_MOESM3_ESM.doc (32 kb)
Supplementary material 3 (DOC 32 kb)


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Authors and Affiliations

  1. 1.Department of Signal Transduction and Biogenic AminesChittaranjan National Cancer InstituteKolkataIndia
  2. 2.Department of Surgical OncologyChittaranjan National Cancer InstituteKolkataIndia
  3. 3.Department of PathologyChittaranjan National Cancer InstituteKolkataIndia
  4. 4.Department of Epidemiology and BiostatisticsChittaranjan National Cancer InstituteKolkataIndia
  5. 5.Department of Molecular PathologyMitra BiotechBengaluruIndia
  6. 6.Department of Cancer BiologyMitra BiotechBengaluruIndia

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