Amemiya K, Kurachi H, Adachi H, Morishige K I, Adachi K, Imai T, Miyake A
Department of Obstetrics and Gynecology, Osaka University Medical School, Japan.
J Endocrinol. 1994 Nov;143(2):291-301. doi: 10.1677/joe.0.1430291.
We have studied the expression of epidermal growth factor (EGF) and EGF receptors (EGF-R) in isolated human trophoblast cells at various stages of differentiation and also the biological significance of the EGF/EGF-R autocrine and paracrine mechanism. Cytotrophoblast cells were isolated from human placental tissues of 6-9 weeks of gestation. Trophoblast cells underwent morphological and functional differentiation during in vitro culture. The expression of EGF and EGF-R protein and mRNA was studied in trophoblast cells cultured for 0-5 days, using immunocytochemical staining, and reverse transcription and polymerase chain reaction. Monoclonal antibodies (mAbs) against EGF and EGF-R showed specific staining in trophoblast cells at all stages of differentiation. Both EGF and EGF-R gene transcripts were detected in RNA samples isolated from trophoblast cells at all stages. These data suggest the presence of an EGF/EGF-R autocrine and paracrine mechanism in human trophoblast cells. Next, we examined the biological significance of this mechanism on trophoblast cell differentiation in vitro. EGF added to the culture medium significantly increased human chorionic gonadotrophin-beta (hCG-beta) secretion and, more importantly, anti-EGF neutralizing mAbs significantly reduced both hCG-beta and human placental lactogen secretion from trophoblast cells in culture. All these results suggest that human trophoblast cells express both EGF and EGF-R, and that EGF may play an important role in the functional differentiation of human trophoblast cells.
我们研究了表皮生长因子（EGF）和EGF受体（EGF-R）在人滋养层细胞不同分化阶段的表达情况，以及EGF/EGF-R自分泌和旁分泌机制的生物学意义。从妊娠6 - 9周的人胎盘组织中分离出细胞滋养层细胞。滋养层细胞在体外培养过程中经历了形态和功能上的分化。使用免疫细胞化学染色、逆转录和聚合酶链反应，研究了培养0 - 5天的滋养层细胞中EGF和EGF-R蛋白及mRNA的表达。抗EGF和EGF-R的单克隆抗体（mAb）在滋养层细胞分化的各个阶段均显示出特异性染色。在从各个阶段的滋养层细胞中分离的RNA样本中均检测到了EGF和EGF-R基因转录本。这些数据表明人滋养层细胞中存在EGF/EGF-R自分泌和旁分泌机制。接下来，我们研究了该机制对体外滋养层细胞分化的生物学意义。向培养基中添加EGF显著增加了人绒毛膜促性腺激素-β（hCG-β）的分泌，更重要的是，抗EGF中和性单克隆抗体显著降低了培养的滋养层细胞中hCG-β和人胎盘催乳素的分泌。所有这些结果表明，人滋养层细胞同时表达EGF和EGF-R，并且EGF可能在人滋养层细胞的功能分化中发挥重要作用。
