Differential display analysis of oxygen-mediated changes in gene expression in first trimester human trophoblast cells.
Physiologic or pathologically induced periods of exposure to relatively low levels of oxygen during pregnancy affect the expression and function of certain genes in the placenta. In this study, the differential display technique was utilized to identify genes that are regulated in cultured cytotrophoblast cells by exposure to low levels of oxygen. Using this approach, four genes, which have been designated HRF-1, HRF-2, HRF-6, and HRF-8, were cloned and partially characterized. Northern blot analysis showed that clones HRF-1 and HRF-2 were downregulated in response to exposure to low levels of oxygen, whereas expression of HRF-6 and HRF-8 was increased. DNA sequencing and sequence analysis revealed that HRF-1 may represent an alternatively spliced or tissue-specific form of the Kruppel family zinc finger protein znfp104 gene. Clone HRF-2 showed a high degree of identity with exons 9, 10 and 11 of N33, a gene that is located within a homozygously deleted region of metastatic prostate cancer. Clones HRF-6 and HRF-8 did not exhibit significant sequence identity with known sequences in GenBank and may represent novel genes. None of these genes have previously been shown to be present in trophoblast cells, nor have their expressions been shown to be regulated by oxygen. This study demonstrates that the differential display technique is a novel and effective method to analyse oxygen-mediated changes in gene expression in trophoblast cells.
Department of Anatomy and Cell Biology, Queen's University, Kingston, Ontario, Canada.
B J
BJ
Pak
H
H
Park
E R
ER
Chang
S C
SC
Pang
C H
CH
Graham
D000328:Adult; D001483:Base Sequence; D015152:Blotting, Northern; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D018257:Helix-Loop-Helix Motifs / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010100:Oxygen / Q000494:pharmacology*; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics; D014158:Transcription, Genetic; D014327:Trophoblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism
D017931:DNA Primers; D012333:RNA, Messenger; D014157:Transcription Factors; D010100:Oxygen