Open AccessErythropoietin receptor mRNA expression in human endothelial cells.
Author(s) -
Athanasius Anagnostou,
Ziyao Liu,
Manfred Steiner,
Kyung Chin,
Eun S. Lee,
Noubar Kessimian,
Constance Tom Noguchi
Publication year - 1994
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.91.9.3974
Subject(s) - erythropoietin receptor , erythropoietin , biology , microbiology and biotechnology , umbilical vein , receptor , endothelial stem cell , messenger rna , cell culture , endocrinology , in vitro , gene , biochemistry , genetics
A previous report demonstrated that endothelial cells have erythropoietin receptors and respond to this hormone with enhanced proliferation. The present study demonstrates the existence of mRNA for erythropoietin receptor in human umbilical vein endothelial cells. We have reverse transcribed mRNA of endothelial cells and then used different PCR primers to amplify erythropoietin receptor target cDNA between exons 5 and 6 as well as 3-5 in addition to an internal standard DNA fragment. Correspondence of size as well as location of restriction endonuclease scission (Ava II) was used in comparing the amplified fragments of human endothelial cell erythropoietin receptor to those of two human erythroleukemia cell lines, OCIM1 and K562. No alpha- or gamma-globin mRNA was detected in endothelial cells but was readily demonstrable in OCIM1 cells. In addition, to determine whether the expression of human erythropoietin receptor on endothelial cells occurs in vivo, sections of umbilical cord and placenta were immunostained with antibodies against the extracellular portion of the receptor; the results showed strong positive staining of the vascular endothelium.