Human umbilical vein endothelial cells express multiple prolactin isoforms.
Members of the prolactin (PRL) hormonal family have direct effects on endothelial cell proliferation, migration and tube formation. Moreover, isoforms of PRL may function as autocrine regulators of endothelial cells. Bovine brain capillary endothelial cells (BBCEC) express the PRL gene, while anti-PRL antibodies inhibit BBCEC proliferation. Here, we show the expression of the PRL gene into various PRL isoforms in endothelial cells from the human umbilical vein. Reverse transcription-polymerase chain reaction of total RNA from human umbilical vein endothelial cells (HUVEC) detected the full-length PRL mRNA as well as a 100 bp smaller PRL transcript similar to the one previously reported in BBCEC. HUVEC were positive to PRL immunocytochemistry. In addition, various PRL immunoreactive proteins were detected in HUVEC extracts and HUVEC conditioned media by metabolic labelling immunoprecipitation analysis. These PRL immunorelated proteins had apparent molecular masses of 60, 23, 21, 16 and 14 kDa. In contrast to previous findings in BBCEC, HUVEC conditioned media contained very little PRL bioactivity as determined by the selective bioassay of Nb2 cell proliferation. Moreover, some polyclonal or monoclonal antibodies directed against PRL stimulated HUVEC proliferation, in contrast to the inhibitory effect seen in BBCEC. The present findings extend the previous observations about the expression of PRL gene in endothelial cells from bovine brain capillaries to human cells of the umbilical vein, implicating that endothelium from different types of vessels and species share the expression of PRL gene but may differ in the putative autocrine role of the PRL isoforms expressed.
Corbacho
A M
AM
Centro de Neurobiología, Universidad Nacional Autónoma de México, 76001, Querétaro, Qro, México.
Macotela
Y
Y
Nava
G
G
Torner
L
L
Dueñas
Z
Z
Noris
G
G
Morales
M A
MA
Martínez De La Escalera
G
G
Clapp
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001681:Biological Assay; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D004730:Endothelium, Vascular / Q000737:chemistry*; D006801:Humans; D007150:Immunohistochemistry; D011233:Precipitin Tests; D011388:Prolactin / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D020033:Protein Isoforms / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013268:Stimulation, Chemical; D014471:Umbilical Veins
D000911:Antibodies, Monoclonal; D017077:Culture Media, Conditioned; D020033:Protein Isoforms; D012333:RNA, Messenger; D011388:Prolactin