Effect of proton beam on blood vessel formation in early developing zebrafish (Danio rerio) embryos.
Proton beam therapy can kill tumor cells while saving normal cells because of its specific energy delivery properties and so is used to various tumor patients. However, the effect of proton beam on angiogenesis in the development of blood vessels has not been determined. Here we used the zebrafish model to determine in vivo whether proton beam inhibits angiogenesis. Flk-1-GFP transgenic embryos irradiated with protons (35 MeV, spread out Bragg peak, SOBP) demonstrated a marked inhibition of embryonic growth and an altered fluorescent blood vessel development in the trunk region. When cells were stained with acridine orange to evaluate DNA damage, the number of green fluorescent cell death spots was increased in trunk regions of irradiated embryos compared to non-irradiated control embryos. Proton beam also significantly increased the cell death rate in human umbilical vein endothelial cells (HUVEC), but pretreatment with N-acetyl cystein (NAC), an antioxidant, reduced the proton-induced cell death rate (p<0.01). Moreover, pretreatment with NAC abrogated the inhibition of trunk vessel development and prevented the trunk malformation caused by proton irradiation. In conclusion, proton irradiation significantly inhibited in vivo vascular development possibly due to increased vascular cell death via reactive oxygen species formation.
School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University, Daegu, 702-701, Korea.
Gun Hyuk
GH
Jang
Ji-Hong
JH
Ha
Tae-Lin
TL
Huh
You Mie
YM
Lee
D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D030801:Animals, Genetically Modified; D000975:Antioxidants / Q000494:pharmacology; D001808:Blood Vessels / Q000187:drug effects / Q000196:embryology / Q000378:metabolism / Q000528:radiation effects*; D016923:Cell Death / Q000528:radiation effects; D002478:Cells, Cultured; D004249:DNA Damage; D004307:Dose-Response Relationship, Radiation; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology / Q000528:radiation effects; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D018919:Neovascularization, Physiologic / Q000187:drug effects / Q000528:radiation effects*; D011522:Protons; D017382:Reactive Oxygen Species / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics / Q000378:metabolism
D000975:Antioxidants; D011522:Protons; D017382:Reactive Oxygen Species; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D040301:Vascular Endothelial Growth Factor Receptor-2; D000111:Acetylcysteine