Irradiation of mechanically-injured human arterial endothelial cells leads to increased gene expression and secretion of inflammatory and growth promoting cytokines.
Radiation therapy is applied to inhibit neointima formation after percutaneous transluminal coronary angioplasty (PTCA). In this study, we evaluated the effect of irradiation on re-endothelialisation of circular denuded tracks made in post-confluent cultures of arterial endothelial cells (ECs) and on cellular factors involved in this process. Image analysis and time-lapse microcinematography revealed cell migration into denuded areas starting 4h after injury. Fifty percent coverage was achieved at 14.8 +/- 2.0 h. Using competitive PCR and flow cytometry techniques, no significant changes in mRNA expression of interleukin-1beta (IL-1beta), interleukin-8 (IL-8), basic fibroblast growth factor (bFGF or FGF-2), transforming growth factor-beta1 (TGF-beta1), platelet-derived growth factor A (PDGF-A), platelet-derived growth factor B (PDGF-B) and tissue factor (TF), and surface molecule expression of anti-intercellular adhesion molecule-1 (ICAM-1), anti-vascular cell adhesion molecule-1 (VCAM-1), anti-platelet/endothelial cell adhesion molecule-1 (PECAM-1), MHC-1, TF and Fas were observed. However, injury did significantly (P < 0.05) elevate the release of IL-8 and FGF-2 protein in the cell culture supernatant, as assessed by ELISA. Radiation (15Gy) given immediately after injury did not affect the kinetics of re-endothelialisation up to 48 h, in spite of the fact that no cell divisions were observed. Thereafter cell density decreased and cultures deteriorated. Compared to cultures exposed to injury alone, radiation induced significant (P < 0.05) increases in mRNA levels of IL-8 (1.35 +/- 0.10-fold increase at 4h), FGF-2 (1.62 +/- 0.10-fold at 4h; 1.76 +/- 0.33-fold at 24h) and IL-1beta (2.76 +/- 0.40-fold at 24h), whereas mRNA levels of TGF-beta1, PDGF-A and PDGF-B increased about 1.2-fold. IL-8 and FGF-2 protein concentrations in the media were higher than those observed in non-irradiated injured cell cultures; however, this difference was not significant. Radiation induced a 2.3 +/- 0.3-fold increase (P < 0.05) in Fas surface expression only. In conclusion, irradiation of mechanically-injured human EC leads to increased gene expression and protein secretion of inflammatory and growth promoting cytokines.
Wondergem
J
J
Department of Clinical Oncology, K1-P, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden, The Netherlands. j.wondergem@lumc.nl
Wedekind
L E
LE
Bart
C I
CI
Chin
A
A
van der Laarse
A
A
Beekhuizen
H
H
D015815:Cell Adhesion Molecules / Q000378:metabolism; D002455:Cell Division / Q000528:radiation effects; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000293:injuries* / Q000378:metabolism / Q000473:pathology / Q000528:radiation effects*; D015870:Gene Expression / Q000528:radiation effects*; D006133:Growth Substances / Q000378:metabolism; D006801:Humans; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis
D015815:Cell Adhesion Molecules; D016207:Cytokines; D006133:Growth Substances; D012333:RNA, Messenger