Analysis of the involvemetnt of CD9 in the regulation of human trophoblast invasion

CD9参与调节人滋养细胞侵袭的分析

• 批准号: 09671674
• 负责人: INOUE Takuya
• 金额: $ 2.24万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671674/
• 关键词: Implantation; CD9; Trophoblast; Invasion; Integrin; Monoclonal Antibody

Trophoblast invasion is one of the most important steps of implantation and placentation. Although many elements of trophoblast invasion resemble the events that occur during malignant tumor cell invasion, trophoblast invasion is confined spatially to the uterus and temporally to early pregnancy. However, the regulatory mechanisms which prevent the extravillous trophoblasts from further invasion are not well understood.The CD9 molecule is a 24-27 kDa cell surface glycoprotein, which may be related to Schwann cell migration and adhesion. In this study, we showed that CD9 was expressed on the extravillous trophoblasts in the cell columns of first trimester placentae, which invade into endometrium during implantation and placentation, but not on villous trophoblasts. Moreover, proteins were purified from chorion leave by affinity chromatography with anti-integrin alpha3 and alpha5 monoclonal antibodies and Western blotting, revealed that CD9 was associated with both integrins. In order to estimate the functional role of CD9 on trophoblasts, we used the human trophoblast-like choriocarcinoma cell line BeWo. Anti-CD9 monoclonal antibody (mAb) showed significant stimulatory effect on BeWo cell invasion. On the other hand, mAbs against integrins alpha3 and alpha5 inhibited BeWo cell invasion. Anti-CD9 mAb also showed a stimulatory effect on BeWo cell invasion under the treatment with mAb against integrin alpha3. In contrast, it had no effect under the treatment with mAb against integrin alpha5.These findings indicate that CD9 is deeply related to the invasive properties of BeWo cells and that the role of CD9 in WeBo cell invasion is partially mediated by integrin alpha5, suggesting the involvement of CD9 in trophoblastic invasion.

滋养细胞的入侵是植入和胎盘的最重要步骤之一。尽管许多滋养细胞侵袭的元素类似于恶性肿瘤细胞入侵期间发生的事件，但滋养细胞的侵袭在空间上局限于子宫，并在时间上限制为早期妊娠。然而，尚不清楚防止牙周滋养细胞进一步侵袭的调节机制。CD9分子是24-27 kDa细胞表面糖蛋白，这可能与Schwann细胞迁移和粘附有关。在这项研究中，我们表明CD9是在妊娠中期胎盘的细胞柱上的跨滋养细胞上表达的，在植入和胎盘植入期间，它们侵入子宫内膜，但在绒毛细胞上不侵入子宫内膜。此外，蛋白质是通过与抗积蛋白α3和α5单克隆抗体和蛋白质印迹的亲和色谱从绒毛膜休假中纯化的，发现CD9与两种整合蛋白有关。为了估计CD9在滋养层细胞上的功能作用，我们使用了人类滋养细胞样的绒毛膜绒毛膜细胞系。抗CD9单克隆抗体（MAB）对BOWO细胞侵袭显示出显着的刺激作用。另一方面，针对整联蛋白α3和α5的mAb抑制了BEWO细胞侵袭。抗CD9 MAB还显示出用MAB对整合蛋白α3治疗的BOWO细胞侵袭的刺激作用。相比之下，在用MAB治疗中对整合蛋白alpha的处理没有影响。这些发现表明CD9与Bewo细胞的侵袭性深度相关，并且CD9在Webo细胞入侵中的作用部分由整合素α5介导，这表明CD9在滋养性粒细胞性侵袭中的参与。

项目成果

Takashi Hirano: "CD9 is involved in invasion of human trophoblast-like choriocarcinoma cell line,BeWo cells" Molecular Human Reproduction.

Takashi Hirano：“CD9 参与人类滋养细胞样绒毛膜癌细胞系 BeWo 细胞的侵袭”《分子人类生殖》。

Hiroshi, Katsuragawa: "Monoclonal Antibody against Phosphatidylserine Inhibits In Vitro Human Trophoblastic Hormone Production and Invasion." Biology of Reproduction. 56. 50-58 (1997)

Hiroshi, Katsurakawa：“抗磷脂酰丝氨酸的单克隆抗体可抑制体外人类滋养细胞激素的产生和侵袭。”

Takeshi, Hirano: "CD9 is expressed in extravillous trophoblasts in association with integrin α3 and integrin α5" Molecular Human Reproduction. 5. 162-167 (1999)

Takeshi, Hirano：“CD9 在绒毛外滋养细胞中表达，与整合素 α3 和整合素 α5 相关”《分子人类生殖》(Molecular Human Reproductive) 5. 162-167 (1999)。

Hirohiko, Watanabe: "Bcl-2 and Fas expression in eutopic and ectopic human endometrium during the menstrual cycle, in relation to endometrial cell apoptosis." American Journal of Obstetrics and Gynecology. 176. 360-367 (1997)

Hirohiko, Watanabe：“月经周期期间在位和异位人类子宫内膜中 Bcl-2 和 Fas 的表达与子宫内膜细胞凋亡的关系。”

Watanabe H,Kanzaki H,Marukawa S,Inoue T,Katsuragawa H,Kaneko Y,Mori T.: "Bcl-2 and Fas expression in eutopic and ectopic human endometrium during the menstrual cycle, in relation to endometrial cell apoptosis." Am J Obstet Gynecol. 176. 360-367 (1997)

Watanabe H、Kanzaki H、Marukawa S、Inoue T、Katsurakawa H、Kaneko Y、Mori T.：“月经周期期间在位和异位人子宫内膜中 Bcl-2 和 Fas 的表达与子宫内膜细胞凋亡的关系。”