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.

滋养层侵袭是着床和胎盘形成的重要环节之一。虽然滋养层细胞侵袭的许多因素类似于恶性肿瘤细胞侵袭过程，但滋养层细胞侵袭在空间上仅限于子宫，在时间上仅限于早期妊娠。然而，阻止绒毛外滋养细胞进一步侵袭的调控机制尚不清楚。CD9分子是一种24-27 kDa的细胞表面糖蛋白，可能与雪旺细胞迁移和黏附有关。在本研究中，我们发现CD9表达于早孕胎盘细胞柱中的绒毛外滋养层细胞，CD9在植入和胎盘形成过程中侵入子宫内膜，但不表达在绒毛滋养层细胞上。此外，用抗整合素α3和α5的单抗亲和层析和Western blotting从绒毛膜叶片中纯化了蛋白质，表明CD9与这两种整合素都是相关的。为了评估CD9对滋养层细胞的功能作用，我们使用了人滋养层细胞样绒癌细胞系BeWo。抗CD9单抗对BeWo细胞的侵袭有明显的刺激作用。另一方面，抗整合素α3和α5的单抗可抑制BeWo细胞的侵袭。在抗整合素α3单抗作用下，抗CD9单抗对BeWo细胞的侵袭也有促进作用。提示CD9与BeWo细胞的侵袭特性密切相关，CD9在Webo细胞侵袭中的作用部分是通过整合素α5介导的，提示CD9参与了滋养细胞的侵袭。

项目成果

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。”

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》。

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。”