HUVEC respond to radiation by inducing the expression of pro-angiogenic microRNAs.

MicroRNAs (miRNAs) represent a class of small non-coding RNAs that control gene expression by targeting mRNAs and triggering either repression of translation or RNA degradation. They have been shown to be involved in a variety of biological processes such as development, differentiation and cell cycle control, but little is known about their involvement in the response to irradiation. We showed here that in human umbilical vein endothelial cells (HUVEC) some miRNAs previously shown to have a crucial role in vascular biology are transiently modulated in response to a clinically relevant dose of ionizing radiation. In particular we identified an early transcriptional induction of several members of the microRNA cluster 17-92 and other microRNAs already known to be related to angiogenesis. At the same time we observed a peculiar behavior of the miR-221/222 cluster, suggesting an important role of these microRNAs in HUVEC homeostasis. We observed an increased efficiency in the formation of capillary-like structures in irradiated HUVEC. These results could lead to a new interpretation of the effect of ionizing radiation on endothelial cells and on the response of tumor endothelial bed cells to radiotherapy.

Low-dose ionizing radiation stimulates transcription and production of endothelin by human vein endothelial cells.

A transient increase of EDN1 mRNA accumulation is observed in human vein endothelial cells (HUVECs) after a low dose of ionizing radiation. The kinetics of this mRNA accumulation parallels that of other AP1-regulated transcripts, showing a sharp peak 2 h after irradiation. This accumulation is followed by a net increase of endothelin 1 and big endothelin 1 in the cytoplasm that reaches a peak 4 h after irradiation. We followed the kinetics of endothelin 1 secretion in cell culture medium and did not find a detectable increase in the rate of secretion by the irradiated cells compared to sham-irradiated cells. We conclude that in HUVEC monolayers, an increase in endothelin production does not automatically correspond to an increase in secretion. These findings suggest that endothelin is an important component in the response of endothelial cells to ionizing radiation and that it could be used as a biomarker for low-dose irradiation of endothelial tissues.

Transcriptional response of human umbilical vein endothelial cells to low doses of ionizing radiation.

We used cDNA microarray hybridization technology to monitor the transcriptional response of Human Umbilical Vein Endothelial (HUVEC) cells to x-rays doses ranging from 2 to 200 cGy. An early time window from irradiation (4h) was selected in order to minimize the effects of the cell cycle blockage eventually induced at high doses of irradiation. Three different gene-clustering algorithms have been used to group the 4134 monitored ORF based on their transcriptional response in function of the irradiation dose. The results show that while few genes exhibit a typical dose-dependent modulation with a variable threshold, most of them have a different modulation pattern, peaking at the two intermediate doses. Strikingly even the lowest dose used (2 cGy) seems to be very effective in transcriptional modulation. These results confirm the physiological relevance of sublethal-dose exposures of endothelial cells and strengthens the hypothesis that alternative dose-specific pathways of radioadaptive response exist in the mammalian cells. 111 genes were found to be modulated at all doses of irradiation. These genes were functionally classified by cellular process or by molecular function. Genes involved in coagulation and peroxidase activity and structural constituent of ribosomes were over-represented among the up-regulated genes as compared with their expected statistical occurrence. Three genes coding for regulatory kinase activities (CDK6; PRCKB1 and TIE) are found down-regulated at all doses of irradiation.