Identification of a stress-induced protein during human trophoblast differentiation by differential display analysis.
Identification of a stress-induced protein during human trophoblast differentiation by differential display analysis.
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DOI：
10.1095/biolreprod61.3.681
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发表时间：
1999-09
影响因子：
3.6
通讯作者：
Bo Xu;Lin Lin-Lin;N. Rote
中科院分区：
文献类型：
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作者：
Bo Xu;Lin Lin-Lin;N. Rote
人胎盘滋养层细胞的分化特征是单核绒毛细胞滋养层融合形成多核合胞体。这一事件与基因表达的巨大变化有关。在本研究中，我们已经采用了一种敏感的方法-差异显示分析，以评估在体外毛喉素诱导分化的人滋养层细胞模型，绒毛膜癌BeWo的基因表达的变化。我们确定了7个基因上调，其表达和功能以前没有报道在滋养层。通过与GenBank数据库的序列比较，发现4个上调基因是新的。其他三个基因编码人细胞色素p450 IIC，肌苷一磷酸脱氢酶II型，还原剂和衣霉素反应蛋白（RTP）。北方印迹分析表明，RTP mRNA的表达诱导BeWo与毛喉素孵育24小时后的3倍，并增加到11倍毛喉素治疗72小时。用原位杂交技术进一步研究RTP在孕中期和足月胎盘组织中的表达模式。RTP mRNA主要表达在合体滋养细胞在中期妊娠和足月胎盘。BeWo细胞中RTP基因的表达具有蛋白激酶C依赖性。这是第一次描述RTP基因在胎盘中的表达，也是第一次阐明RTP基因表达调控的信号通路。这些结果表明RTP可能在滋养层细胞增殖和分化中起作用。
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.