Isolation and characterization of a human amnion epithelial cell line that expresses the pregnancy-specific beta 1-glycoprotein gene.
Human pregnancy-specific beta 1-glycoprotein (PSG) is a family of closely related glycoproteins of 72K, 64K, 62K, and 54K. Together with the carcinoembryonic antigen, they form new members of the immunoglobulin superfamily. To study the molecular mechanisms that regulate expression of the PSG gene, we established a human amnion cell line, HAA58OD-8C, immortalized with an origin-defective simian virus-40 (SV40) temperature-sensitive A58 mutant virus. HAA58OD-8C cells were temperature sensitive for maintenance of transformation and expressed genes encoding PSG and the alpha- and beta-subunits of hCG. At the permissive temperature (33 C; transformed phenotype), they expressed low levels of PSG, hCG alpha, and hCG beta mRNAs and synthesized low levels of a 48K PSG polypeptide. At the nonpermissive temperature (39.5 C), HAA58OD-8C cells exhibited a differentiated phenotype, expressed increased levels of PSG, hCG alpha, and hCG beta mRNAs, and produced high levels of PSG polypeptides of 72K and 48K. Sodium butyrate induced PSG mRNA expression, and in the presence of butyrate, HAA58OD-8C cells produced high amounts of PSG polypeptides of 72K, 62K, and 48K. Ribonuclease protection analysis indicated that similar PSG transcripts were expressed by HAA58OD-8C cells and human term placenta. However, these amnion cells expressed selectively a certain population of PSG transcripts. Our results show that this amnion cell line provides a suitable model for studies of PSG gene expression and regulation.
Plouzek
C A
CA
Human Genetics Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892.
Chou
J Y
JY
D000650:Amnion / Q000378:metabolism* / Q000648:ultrastructure; D002460:Cell Line; D002469:Cell Separation; D006063:Chorionic Gonadotropin / Q000235:genetics; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure; D005260:Female; D015870:Gene Expression; D005805:Genes, MHC Class I / Q000235:genetics; D006680:HLA Antigens / Q000235:genetics; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I / Q000235:genetics; D006801:Humans; D008854:Microscopy, Electron; D009693:Nucleic Acid Hybridization; D011247:Pregnancy; D011268:Pregnancy-Specific beta 1-Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012260:Ribonucleases; D013696:Temperature
D006063:Chorionic Gonadotropin; D006680:HLA Antigens; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I; D011268:Pregnancy-Specific beta 1-Glycoproteins; D012333:RNA, Messenger; D012260:Ribonucleases