Identification of a stress-induced protein during human trophoblast differentiation by differential display analysis.
Differentiation of human placental trophoblast is characterized by a process during which mononuclear villous cytotrophoblasts fuse to form a multinucleate syncytium. This event is associated with dramatic changes in gene expression. In the present study, we have applied a sensitive approach-differential display analysis-to evaluate changes in gene expression during in vitro forskolin-induced differentiation of a model of human trophoblast, the choriocarcinoma BeWo. We identified seven genes that were up-regulated; their expression and function have not previously been reported in trophoblast. Four up-regulated genes were novel upon comparison of their sequences to the GenBank database. The other three genes encode human cytochrome p450 IIC, inosine monophosphate dehydrogenase type II, and reducing agent and tunicamycin-responsive protein (RTP). Northern blot analysis revealed that RTP mRNA expression was induced to 3-fold in BeWo after 24-h incubation with forskolin and increased up to 11-fold by 72 h of forskolin treatment. The expression pattern of RTP was further investigated by in situ hybridization on second trimester and term placenta tissues. RTP mRNA was predominantly expressed in syncytiotrophoblasts in both second trimester and term placentae. The expression of RTP gene in BeWo cells was protein kinase C dependent. This is the first description of RTP gene expression in placenta and the first study elucidating the signaling pathway involved in the regulation of RTP gene expression. These results suggest that RTP may play a role in trophoblast cell proliferation and differentiation.
Xu
B
B
Department of Microbiology & Immunology, Wright State University School of Medicine, Dayton, Ohio 45435, USA.
Lin
L
L
Rote
N S
NS
D015152:Blotting, Northern; D002454:Cell Differentiation / Q000235:genetics*; D002455:Cell Division; D002822:Choriocarcinoma; D005576:Colforsin / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D017403:In Situ Hybridization; D010920:Placenta / Q000737:chemistry; D011506:Proteins / Q000235:genetics*; D012313:RNA / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D015398:Signal Transduction; D014327:Trophoblasts / Q000166:cytology* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D011506:Proteins; D012333:RNA, Messenger; C118748:reducing agent and tunicamycin-responsive protein, human; D005576:Colforsin; D012313:RNA