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Updated: Mar 25, 2026
Optimization of a Multiplex RNA-based Expression Assay Using Breast Cancer Archival Material
Published on: August 1, 2018
EPH/ephrin profile and EPHB2 expression predicts patient survival in breast cancer
Anna-Maria Husa1,2, Željana Magić1, Malin Larsson3
1Department of Clinical and Experimental Medicine, Division of Oncology, Linköping University, Linköping, Sweden.
Abstract:
The EPH and ephrins function as both receptor and ligands and the output on their complex signaling is currently investigated in cancer. Previous work shows that some EPH family members have clinical value in breast cancer, suggesting that this family could be a source of novel clinical targets. Here we quantified the mRNA expression levels of EPH receptors and their ligands, ephrins, in 65 node positive breast cancer samples by RT-PCR with TaqMan® Micro Fluidics Cards Microarray. Upon hierarchical clustering of the mRNA expression levels, we identified a subgroup of patients with high expression, and poor clinical outcome. EPHA2, EPHA4, EFNB1, EFNB2, EPHB2 and EPHB6 were significantly correlated with the cluster groups and particularly EPHB2 was an independent prognostic factor in multivariate analysis and in four public databases. The EPHB2 protein expression was also analyzed by immunohistochemistry in paraffin embedded material (cohort 2). EPHB2 was detected in the membrane and cytoplasmic cell compartments and there was an inverse correlation between membranous and cytoplasmic EPHB2. Membranous EPHB2 predicted longer breast cancer survival in both univariate and multivariate analysis while cytoplasmic EPHB2 indicated shorter breast cancer survival in univariate analysis. Concluding: the EPH/EFN cluster analysis revealed that high EPH/EFN mRNA expression is an independent prognostic factor for poor survival. Especially EPHB2 predicted poor breast cancer survival in several materials and EPHB2 protein expression has also prognostic value depending on cell localization.
Insights
High expression of EPH receptors and ephrins (EFN) correlates with poor breast cancer survival. Specifically, EPHB2 expression, depending on its cellular location, serves as a significant prognostic factor for patient outcomes.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- The EPH receptor tyrosine kinase and ephrin ligand system plays a crucial role in cellular signaling, with emerging implications in cancer biology.
- Previous research indicates the clinical relevance of certain EPH family members in breast cancer, highlighting their potential as therapeutic targets.
Purpose of the Study:
- To investigate the mRNA expression levels of EPH receptors and ephrins in node-positive breast cancer samples.
- To identify correlations between EPH/ephrin expression patterns and clinical outcomes, particularly patient survival.
- To evaluate the prognostic value of EPHB2, a specific EPH family member, at both mRNA and protein levels.
Main Methods:
- Quantitative mRNA expression analysis using RT-PCR with TaqMan® Micro Fluidics Cards Microarray in 65 node-positive breast cancer samples.
- Hierarchical clustering to identify patient subgroups based on EPH/ephrin mRNA expression levels.
- Immunohistochemistry to assess EPHB2 protein expression and localization in a separate cohort of paraffin-embedded breast cancer tissues.
Main Results:
- A subgroup of patients with high EPH/ephrin mRNA expression exhibited a poor clinical outcome.
- EPHA2, EPHA4, EFNB1, EFNB2, EPHB2, and EPHB6 mRNA levels were significantly correlated with identified patient clusters.
- EPHB2 emerged as an independent prognostic factor for poor survival in multivariate analysis and public databases.
- Membranous EPHB2 protein expression correlated with longer survival, whereas cytoplasmic EPHB2 indicated shorter survival.
Conclusions:
- EPH/ephrin cluster analysis demonstrates that high mRNA expression is an independent prognostic indicator of poor survival in breast cancer.
- EPHB2 is a key predictor of poor breast cancer survival, with its prognostic value influenced by its subcellular localization (membranous vs. cytoplasmic).
- The study underscores the potential of targeting the EPH/ephrin pathway for novel breast cancer therapies.
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