Patients receiving recombinant human erythropoietin (rHuEPO) may experience side effects arising from the vascular system. The underlying mechanisms, how-ever, are largely unknown.
Methods
To elucidate downstream events following eryth-ropoietin receptor triggering, a differential display analysis of human vascular endothelial cell mRNA was performed.
Results
We identified eight genes that were upregulated by rHuEPO as confirmed in two further independent cell culture experiments using a semiquantitative reverse tran-scriptase polymerase chain reaction (RT-PCR) protocol. The genes coded for proteins that may be assigned to four differ-ent groups: 1) proteins implicated in the regulation of vas-cular functions (thrombospondin-1, 20 kDa myosin regula-tory light chain; relative increase of rHuEPO induced mRNA levels: 155.2%, P=0.043; 137.6%, P=0.046, respectively); 2) gene products involved in gene transcription and/or transla-tion (c-myc purine-binding transcription factor PuF, trypto-phanyl-tRNA synthetase, S19 ribosomal protein; increase of mRNA levels: 126.4%, P=0.032; 150.9%, P=0.012; 134.9%, P=0.038); 3) subunits of mitochondrial enzymes related to energy transfer (NADH dehydrogenase subunit 6, cyto-chrome C oxidase subunit 1; increase of mRNA concentra-tions: 141.7%, P=0.007; 140.3%, P=0.01); and 4) regulators of signal transduction (protein tyrosine phosphatase G1, in-crease of transcript level: 160.3%, P=0.016).
Conclusions
We report on novel molecular downstream events following rHuEPO receptor triggering of human vas-cular endothelial cells. We identified EPO-responsive imme-diate-early genes, coding for proteins involved in vascular functions, gene transcription, and/or translation, energy transfer, and signal transduction. Thus, our data provide new insights into the molecular changes induced by EPO in human vascular endothelial cells.
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