ABSTRACT
BACKGROUND: There is accumulating evidence from preclinical studies that circulating endothelial cells (CECs) play an important role in neovascularization and tumor growth. The role of CECs in human cancer progression is sparsely investigated. We therefore analyzed CECs in peripheral blood of cancer patients. In addition, we correlated CEC levels in these patients with plasma levels of cytokines that are known to mobilize CECs in experimental models. PATIENTS AND METHODS: Viable CECs were isolated, quantified and cultured from cancer patients' whole blood by using magnetic beads coupled to an antibody directed against CD146, a pan-endothelial marker. Viable cells were visualized by calceinAM staining. Positive staining for specific endothelial cell markers [i.e. von Willebrand factor, CD31, vascular endothelial cell growth factor (VEGF) receptor-2] was used to confirm the endothelial phenotype. RESULTS: Cancer patients with progressive disease (95 patients) had on average 3.6-fold more CECs than healthy subjects (46 patients, P <0.001). Patients (17) with stable disease had CEC numbers equal to that circulating in healthy subjects (P = 0.69). A subset of in vitro cultured CECs incorporated into endothelial layers and formed colonies. Plasma levels of cytokines that are thought to mobilize CECs from the bone marrow [VEGF, placental growth factor, stromal cell derived factor 1alpha and stem cell factor (71 patients)] did not correlate with CEC amounts. The levels of viable CECs in cancer patients were modified by granulocyte colony-stimulating factor treatment and chemotherapy. CONCLUSION: In progressive cancer patients, the amount of CECs is increased. These CECs are viable and may contribute to vessel formation. The number of CECs is influenced by anticancer treatment.
Subject(s)
Biomarkers, Tumor/analysis , Endothelium, Vascular/cytology , Neoplasms/blood supply , Neoplasms/pathology , Angiogenesis Inhibitors , Cytokines/blood , Disease Progression , Endothelial Cells/pathology , Endothelial Growth Factors/blood , Endothelium, Vascular/pathology , Fluorescent Antibody Technique , Follow-Up Studies , Humans , Neovascularization, Pathologic/drug therapy , Neovascularization, Pathologic/pathology , Vascular Endothelial Growth Factor A , von Willebrand Factor/analysisABSTRACT
BACKGROUND: Tumour-induced microvascular networks have become attractive targets in cancer therapy. Strategies that target both tumour cells and vasculature have not been investigated in models of early metastatic colorectal disease. The efficacy of a combination of conventional chemotherapy with a potent angiogenesis inhibitor (endostatin or angiostatin) in a murine model of early colorectal liver metastasis was studied. METHODS: Sixty-six mice were subjected to intrasplenic injection of C26 tumour cells to induce colorectal liver metastases. Control animals received phosphate-buffered saline (n = 8) or citrate buffer (n = 8). Treatment included conventional chemotherapy (n = 9), endostatin (n = 8), high-dose (n = 5) or low-dose (one-tenth of optimal dose; n = 10) angiostatin, as well as the combination of either of these drugs with chemotherapy (n > 5). Clinical appearance was scored daily using a semiquantitative scale. Liver weight, macroscopic and histological tumour involvement (hepatic replacement area; HRA) were measured upon death at day 12. RESULTS: Treated mice displayed significantly better clinical scores than controls, except for those animals treated with low-dose angiostatin with or without chemotherapy. Treatment with conventional chemotherapy resulted in a decrease in HRA from 42.3 to 29.1 per cent (P < 0.001). The addition of angiostatin or endostatin to conventional chemotherapy improved antitumoral efficacy, in a multiplicative manner, resulting in a HRA of approximately 3.5 per cent (P < 0.001). CONCLUSION: The addition of angiostatin or endostatin to conventional chemotherapy enhanced antitumoral efficacy in a murine model of early colorectal liver metastasis.
