Investigation of controlled human extravillous trophoblast invasion

人类滋养层细胞分化为两种主要细胞系：绒毛细胞和绒毛外细胞。绒毛状滋养层覆盖在胎盘的绒毛膜绒毛上，其功能是将氧气和营养物质从母体输送给胎儿。相反，绒毛外滋养层细胞（EVT）侵入母体子宫，并沿螺旋动脉向上迁移，将其转化为低阻力的大血管。EVT侵袭的许多方面与肿瘤细胞侵袭期间发生的事件相似。然而，与肿瘤侵袭不同，EVT侵袭是精确调节的，在空间上局限于子宫，在时间上局限于妊娠早期。由于EVT受控制的侵袭机制尚未完全阐明，因此本项目首先通过多种方法鉴定EVT特异表达的分子。此后，我们检测了每种分子在人滋养层细胞中的作用，特别是在它们的受控侵袭中。 ...更多信息 nt EVT），以鉴定EVT中表达的新分子，获得了几个与EVT反应的mAb克隆，并通过氨基酸序列测定、cDNA淘选或免疫筛选等方法对它们的抗原进行了分析。这些分析表明，整联蛋白α 5，细胞角蛋白18和黑素瘤细胞粘附分子（MCAM）在EVT中表达。此外，mAb 5-23识别未知的200 kD细胞表面蛋白。除了mAb分析外，我们进行了cDNA微阵列分析，以确定在非肿瘤组织中差异表达的基因，妊娠晚期绒毛膜层侵入性EVT与妊娠早期胎盘仍有侵入性EVT比较。鉴定了几个功能未知的表达序列标签（EST）。其中EST 9主要表达于妊娠早期胎盘组织中，经5 'RACE分析，EST 9基因全长为6 kb，编码40 kD的转录因子。EST 9在人类滋养层细胞中的功能正在研究中。少

Human trophoblasts differentiate into two main cell lineages : villous and extravillous cells. Villous trophoblasts cover the chorionic villi of the placenta, their function being to transport oxygen and nutrients from the mother to the fetus. In contrast, extravillous trophoblasts (EVT) invade maternal uterus and also migrate up the spiral arteries, transforming them into large vessels of low resistance. Many aspects of EVT invasion are similar to events that occur during tumor cell invasion. However, unlike tumor invasion, EVT invasion is precisely regulated, confined spatially to the uterus and temporally to early pregnancy. The mechanism of controlled EVT invasion is not clarified well.Therefore in this project, we first identified molecules which specifically express in EVT by several methods. Thereafter, we examined the role of each molecule in human trophoblasts, especially in their controlled invasiveness.We raised mouse mAb against human term chorion laeve (its contains abunda … More nt EVT) to identify novel molecules which are expressed in EVT.Several mAb clones which react with EVT were obtained, and their antigens were analyzed by amino acid sequencing, cDNA panning, or by immunoscreening method. These analyses revealed that integrin alpha 5, cytokeratin 18, and melanoma cell adhesion molecule (MCAM) are expressed in EVT.In addition, mAb 5-23 recognized the unknown cell surface protein of 200 kD.Besides mAb analyses, we carried out cDNA microarray analysis to identify genes which is differently expressed in non-invasive EVT of 3rd trimester chorion laeve compared with still invasive EVT of 1st trimester placenta. Several expressed sequence tag (EST) of unknown function were identified. Among these EST, EST9 was predominantly expressed in 1st trimester placenta which contains invasive EVT.Full length cDNA was determined by 5'RACE, and resultant EST9 mRNA of 6kb encoded a putative transcription factor of 40kD protein. Functions of EST9 in human trophoblast are now under investigation. Less