Ogura K, Sakata M, Okamoto Y, Yasui Y, Tadokoro C, Yoshimoto Y, Yamaguchi M, Kurachi H, Maeda T, Murata Y
Department of Obstetrics and Gynecology, Osaka University Faculty of Medicine, 2-2 Yamadaoka Suita Osaka, 565-0871 Japan.
J Endocrinol. 2000 Feb;164(2):171-8. doi: 10.1677/joe.0.1640171.
Facilitative glucose transporter-1 (GLUT1) is abundant in trophoblast cells and is responsible for glucose transport in the placenta. However, the change in GLUT expression in human placenta upon trophoblast differentiation remains to be clarified. Therefore, we first examined the localization of GLUT1 and GLUT3 using human first-trimester chorionic villi. We found that GLUT1 and GLUT3 were mainly localized to syncytiotrophoblast and cytotrophoblast cells respectively. We analyzed whether placental GLUT1 and GLUT3 expression changes during differentiation using a human choriocarcinoma (BeWo) cell line which is known to show functional and morphological differentiation in response to cAMP in culture. Treatment of BeWo cells with 8-bromo-cyclicAMP (8-bromo-cAMP) increased the level of hCG secretion and induced cell fusion leading to the formation of large syncytia. Treatment of BeWo cells with 8-bromo-cAMP also resulted in a significant increase in glucose uptake on days 2-3 of culture. The stimulating effect of 8-bromo-cAMP on glucose uptake was concentration dependent. Northern and immunoblot analyses revealed that the levels of mRNA and protein of GLUT1, but not of GLUT3, were significantly increased by 8-bromo-cAMP. These findings suggest that 8-bromo-cAMP stimulates GLUT1 expression with differentiation in BeWo cells.
易化葡萄糖转运蛋白1(GLUT1)在滋养层细胞中含量丰富,负责胎盘内的葡萄糖转运。然而,人胎盘在滋养层细胞分化时GLUT表达的变化仍有待阐明。因此,我们首先利用人孕早期绒毛膜绒毛检测了GLUT1和GLUT3的定位。我们发现GLUT1和GLUT3分别主要定位于合体滋养层细胞和细胞滋养层细胞。我们使用人绒毛膜癌细胞系(BeWo)分析了胎盘GLUT1和GLUT3在分化过程中的表达变化,已知该细胞系在培养中对cAMP有功能和形态学上的分化反应。用8-溴环磷酸腺苷(8-bromo-cAMP)处理BeWo细胞可增加hCG分泌水平并诱导细胞融合,导致形成大的合体细胞。用8-溴环磷酸腺苷处理BeWo细胞在培养的第2-3天也导致葡萄糖摄取显著增加。8-溴环磷酸腺苷对葡萄糖摄取的刺激作用呈浓度依赖性。Northern印迹和免疫印迹分析显示,8-溴环磷酸腺苷可显著增加GLUT1的mRNA和蛋白水平,但不影响GLUT3。这些发现表明,8-溴环磷酸腺苷在BeWo细胞分化过程中刺激GLUT1表达。