First-trimester trophoblast cell model gene response to hypoxia.

Abstract

Trophoblast invasion, which sets the stage for placentation and pregnancy outcome, likely occurs in a hypoxic environment. We used microarray technology in a trophoblast cell line to identify hypoxia-responsive genes that may impact placentation. An immortalized extravillous cytotrophoblast cell line, HTR-8/SVneo, was exposed to normoxia (20% oxygen) or hypoxia (1% oxygen) for 6 hours. Total RNA was harvested and prepared for microarray study. Quantitative reverse transcriptase polymerase chain reaction was performed for array confirmation. We confirmed the up- and down-regulation of 10 hypoxia-responsive genes using quantitative reverse transcriptase polymerase chain reaction. Ontologic gene categories that were found to be hypoxia-responsive included motility/migration, angiogenesis, and apoptosis. Specific genes that were found to be up-regulated in this first-trimester array (such as plasminogen activator inhibitor-1 and tissue inhibitor of metalloproteinase 3) have been described in preeclampsia. The hypoxia-responsive genes that we identified may be physiologic in early pregnancy. However, up-regulation of these same genes in later pregnancy augurs poorly.