Arterio-venous gradients of IL-6, plasma and serum VEGF and D-dimers in human cancer

Abstract

The circulating angiogenic factors vascular endothelial growth factor-A, interleukin-6 and the fibrin D-dimer fragment were measured in the mesenteric vein, the uterine vein, as well as in peripheral venous and arterial samples in 21 randomly selected patients with operable colorectal, ovarian and cervical carcinoma. In addition, immunohistochemistry for vascular endothelial growth factor-A and interleukin-6 was performed on colorectal tumours of such patients. Serum and plasma vascular endothelial growth factor-A were not significantly elevated in the vein draining the tumours, despite tumour cell expression of vascular endothelial growth factor-A. Serum vascular endothelial growth factor-A is therefore not all tumour-derived. In contrast, serum interleukin-6 was highly elevated in the draining veins in agreement with expression of interleukin-6 in the cytoplasm of tumour cells. In the megakaryoblastic cell line MEG-01, the expression of vascular endothelial growth factor-A was found to be regulated by interleukin-6. Thus, the higher platelet vascular endothelial growth factor-A load resulting in higher serum vascular endothelial growth factor levels in cancer patients may partly result from an interleukin-6 mediated up-regulation of the expression of vascular endothelial growth factor-A in the precursor of the platelet, i.e. the megakaryocyte. We also confirmed by immunohistochemistry that platelets adhere and aggregate on tumour endothelium. We propose that interleukin-6 indirectly promotes tumour angiogenesis through its up-regulation of the vascular endothelial growth factor-A load in platelets. In addition, the correlations found between peripheral venous interleukin-6 and peripheral venous fibrinogen and D-dimers levels, and the high D-dimer levels found in the draining vein of the tumour, in agreement with fibrin deposits found in the tumour stroma, suggest an important role for interleukin-6 in extra-vascular fibrinogen metabolism. Our results suggest a pivotal role for interleukin-6 in the intrinsic link between haemostasis and angiogenesis. This might be of importance in the development of anti-angiogenic agents based on interference with haemostasis.

Figure 1

(A) VEGF-immunoreactivity in colorectal tumour cells (arrows ×400). (B) Interleukin-6-immunoreactivity in colorectal tumour cells (arrow ×400). (C) Adherence, aggregation and extravasation of platelets (thin arrows) in the vicinity of colorectal tumour cells (thick arrows ×400). (D) Intra-vascular immunostaining of fibrin on endothelial cells (arrow ×400). (E) Extra-vascular stromal staining (thin arrows) of fibrin near tumour cells (thick arrow ×400).

Figure 2

Observed gradients between arterial samples (art.) and tumour efferent blood (EV: efferent vene: mesenteric and uterine vene) for (A) serum VEGF-A, (B) plasma VEGF-A, (C) Interleukin-6 and (D) D-dimers in patients with localised cervical, ovarian and colorectal cancer (n=15).

Figure 3

Observed correlation analysis between peripheral venous serum IL-6 and (A) platelets, (B) D-Dimers and (C) fibrinogen (n=15).

Figure 4

Cell culture experiment in which VEGF-A is measured with ELISA in the conditioned medium of MEG-01 cell lines treated with a blocking antibody against the IL-6 receptor.