Endothelial cell colony forming units derived from malignant breast diseases are resistant to tumor necrosis factor-α-induced apoptosis.

Chen-Pin Chou,Ya-Wen Chen,Huay-Ben Pan,Yu-Lin Chen,Hui-Hwa Tseng,Yi-Chen Yen,Ssu-Han Wang,Shih Sheng Jiang,Yu-Ting Hung

doi:10.1038/srep37450

Abstract

Mobilisation of endothelial progenitor cells (EPCs) from the bone marrow is a crucial step in the formation of de novo blood vessels, and levels of peripheral blood EPCs have been shown to be elevated in certain malignant states. Using flow cytometry and a Hill-based colony forming unit (CFU) assay, the present study indicated that higher levels of CD34 and vascular endothelial growth factor receptor 2 (VEGFR2) double-positive EPCs, as well as increased formation of endothelial cell colony-forming units (EC-CFUs) are associated with benign and malignant breast diseases, providing possible indicators for breast disease detection. Gene expression profiles revealed a genetic difference between CD34+ VEGFR2+ EPCs and EC-CFUs. Decreased expression of tumour necrosis factor receptor 2 (TNFR2) signalling-related genes and inhibition of tumour necrosis factor (TNF)-induced signalling were demonstrated in EC-CFUs derived from patients with malignant breast disease in comparison with those from healthy controls. Interestingly, our data provided the first evidence that EC-CFUs derived from patients with malignant breast disease were resistant to TNF-α-induced apoptosis, indicating a plausible target for future therapeutic interventions.

Highlights

Primitive haemangioblasts to more differentiated endothelial cells[10]
Higher numbers of CD34+ VEGFR2+ endothelial progenitor cells were observed in the peripheral blood of patients with benign and malignant breast diseases
Based on recent studies that have demonstrated circulating endothelial precursor cells (EPCs) using flow cytometry[8,24,25], the numbers of CD34+ VEGFR2+ double-positive cells were first determined in the peripheral blood of breast disease patients and healthy controls (Fig. 1A)

Summary

Introduction

Primitive haemangioblasts to more differentiated endothelial cells[10]. The EPCs were further determined to be included in the population of CD34+/CD133+ progenitor cells. Hill et al used a short-term culture assay with peripheral blood mononuclear cells grown in fibronectin-coated wells to quantify EPCs in men with a spectrum of cardiovascular risk and endothelial functions[2]. With this assay, the circulating levels of angioblast-like EPCs can be quantified by their ability to form an island-like colony called a designated endothelial cell colony-forming unit (EC-CFU). Given the uncertainty in defining phenotypes using cellular markers or the EC-CFU assay, in the present study we aimed to determine whether the genotypic characterization of the EC-CFUs and changes in their gene expression pattern would provide insight into endothelial function and the EPC differentiation capacity between malignant breast diseases and healthy subjects. This was achieved using transcriptomic analysis and validated in vitro to distinguish the detailed molecular fingerprints of the two EC-CFUs

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Conclusion