title: Effects of netrin-1 and netrin-1 knockdown on human umbilical vein endothelial cells and angiogenesis of rat placenta.
abstract: Angiogenesis is an important process essential for the development of placenta. Netrin-1 was first discovered in nervous system and was later found to play roles in angiogenesis. In order to better understand the functional relevance of netrin-1 in placental angiogenesis, we investigated the effect of netrin-1 on human umbilical vein endothelial cells (HUVECs) and rat placenta by employing up-regulation and down-regulation strategies. HUVECs and rat placenta were treated with recombinant netrin-1, and netrin-1 expression in the cells and placenta was reduced by short hairpin RNA (shRNA) in vitro and in vivo. The inhibition efficiency was determined by real-time quantitative polymerase chain reaction (RT-PCR) and Western blotting. The expression of netrin-1 was immunohistochemically located. The results demonstrated that netrin-1 promoted viability, proliferation, migration and tube formation of HUVECs. A strong reduction in cell capability was observed in vitro after netrin-1 expression was inhibited with shRNA. Netrin-1 accelerated neovascularization of placenta in pregnant rats. Suppression of netrin-1 expression in placenta resulted in reduced vascular sprouting in vivo. These findings suggest that netrin-1 is essential for the proper functioning of HUVECs and angiogenesis of rat placenta, and it is involved in the development of placenta and fetus. The proangiogenic effect of netrin-1 might offer an alternative therapeutic approach for the treatment of vascular disease of placenta.
authors.affiliation: Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
authors.forename: H
authors.initials: H
authors.lastname: Xie
authors.forename: L
authors.initials: L
authors.lastname: Zou
authors.forename: J
authors.initials: J
authors.lastname: Zhu
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
mesh_terms: D000818:Animals; D002465:Cell Movement / Q000187:drug effects; D015536:Down-Regulation; D042783:Endothelial Cells / Q000187:drug effects / Q000502:physiology*; D005260:Female; D055785:Gene Knockdown Techniques; D006801:Humans; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D009414:Nerve Growth Factors / Q000037:antagonists & inhibitors / Q000502:physiology*; D000075388:Netrin-1; D010920:Placenta / Q000098:blood supply*; D011247:Pregnancy; D034741:RNA, Small Interfering / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D014471:Umbilical Veins / Q000166:cytology; D015854:Up-Regulation
chemical_list: C000617351:NTN1 protein, human; D009414:Nerve Growth Factors; C000617354:Ntn1 protein, rat; D034741:RNA, Small Interfering; D025521:Tumor Suppressor Proteins; D000075388:Netrin-1