title: The expression of Argonaute2 and related microRNA biogenesis proteins in normal and hypoxic trophoblasts.
abstract: Endogenous microRNAs (miRNAs) post-transcriptionally regulate mRNA and protein expression during tissue development and function. Whereas adaptation to environmental insults are tightly regulated in human tissues, the role of miRNAs and miRNA biogenesis proteins in this context is inadequately explored. We sought to analyse the expression of the key RNAi enzyme Argonaute2 (Ago2) and other miRNA biogenesis proteins in human trophoblasts during differentiation and in hypoxic environment. Using an in vitro analysis of primary term human trophoblasts, we identified the expression of the core miRNA biogenesis proteins in human villous trophoblasts, with expression levels unaffected by cellular differentiation. We found that the miRNA biosynthetic pathway was functional and produced miRNAs, with miR-93 up-regulated and miR-424 down-regulated in hypoxic environment. In contrast, hypoxia did not alter the expression of key miRNA machinery proteins. The pivotal miRNA processing enzyme Ago2, along with its interacting protein DP103, were expressed in normal placentas as well as in placentas from pregnancies complicated by placental hypoperfusion that resulted in fetal growth restriction. Ago2 and DP103 co-immunoprecipitated, and did not limit trophoblast response to hypoxic stress. We concluded that the core miRNA machinery proteins are expressed and functional in human trophoblasts. The influence of hypoxia on the expression of a subset of placental miRNA species is unlikely to reflect altered expression of key miRNA biogenesis proteins.
authors.affiliation: Department of Obstetrics and Gynecology, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: Rogier B
authors.initials: RB
authors.lastname: Donker
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Mouillet
authors.forename: D Michael
authors.initials: DM
authors.lastname: Nelson
authors.forename: Yoel
authors.initials: Y
authors.lastname: Sadovsky
mesh_terms: D060565:Argonaute Proteins; D015687:Cell Hypoxia / Q000235:genetics; D002478:Cells, Cultured; D055541:DEAD Box Protein 20; D053487:DEAD-box RNA Helicases / Q000378:metabolism; D015852:Eukaryotic Initiation Factor-2; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D010448:Peptide Initiation Factors / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034743:RNA-Induced Silencing Complex / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014327:Trophoblasts / Q000378:metabolism*
chemical_list: C466422:AGO2 protein, human; D060565:Argonaute Proteins; D015852:Eukaryotic Initiation Factor-2; D035683:MicroRNAs; D010448:Peptide Initiation Factors; D012333:RNA, Messenger; D034743:RNA-Induced Silencing Complex; C120168:DDX20 protein, human; D055541:DEAD Box Protein 20; D053487:DEAD-box RNA Helicases