Microarray-based identification of differentially expressed genes in hypoxic term human trophoblasts and in placental villi of pregnancies with growth restricted fetuses.

Hypoxia adversely influences the function of the human placenta. We sought to identify a set of hypoxia-regulated transcripts in both term human trophoblasts in vitro and in villous trophoblasts in vivo. Using high-density oligonucleotide microarrays we initially examined differences in gene expression between trophoblast cultured in standard conditions (20% oxygen) vs. hypoxic conditions (< or =1% oxygen), as well as in placental tissues from pregnancies complicated by intrauterine growth restriction vs. matched controls. We used a novel computation method to compile data from the two approaches and identify transcripts that exhibited a marked expression change. Using quantitative PCR we confirmed an up-regulation of transcripts for vascular endothelial growth factor, connective tissue growth factor, follistatin-related protein, N-Myc down-regulated gene 1 and adipophilin in hypoxic term trophoblasts. In contrast, the expression of human placental lactogen and Beckwith-Wiedemann region 1 C was reduced in hypoxic trophoblast. Using in situ hybridization we validated the expression of each transcript in cultured term villous trophoblasts, and determined transcript expression in placental samples derived from four sets of dichorionic twins complicated by growth restriction of one twin. The identification of hypoxic trophoblast signature transcripts may implicate new mediators in pathways underlying trophoblast hypoxic injury and adaptation.