Effects of 3,5,3′-triiodothyronine on the invasive potential and the expression of integrins and matrix metalloproteinases in cultured early placental extravillous trophoblasts
Effects of 3,5,3′-triiodothyronine on the invasive potential and the expression of integrins and matrix metalloproteinases in cultured early placental extravillous trophoblasts
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DOI：
10.1210/jc.2004-0352
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发表时间：
2004-10-01
影响因子：
5.8
通讯作者：
Maruo, T
中科院分区：
文献类型：
--
作者：
Oki, N;Matsuo, H;Maruo, T
众所周知，T(3)通过诱导绒毛滋养细胞的内分泌功能，在维持早期妊娠中发挥着至关重要的作用。然而，T(3) 对绒毛外滋养层 (EVT) 功能的影响仍有待阐明。为了研究T（3）在调节EVT侵袭蜕膜中的可能作用，我们检查了T（3）是否影响EVT侵袭潜力以及培养的早期胎盘EVT中基质金属蛋白酶-2（MMP-2）、MMP-3、组织抑制剂金属蛋白酶-1、胎儿纤连蛋白（FN）和整合素α（5）β（1）的表达。使用人 FN 预包被的培养皿和 FN 预包被的基质胶 Transwell，通过锚定绒毛膜绒毛的酶消化来分离和纯化分化为 EVT 的滋养层。将附着在培养皿上的细胞在补充有10%胎牛血清的DMEM中传代培养48小时，并在24小时传代培养后通过RT-PCR分析进行表征，并在48小时传代培养后通过免疫细胞化学分析进行特异性EVT标记物的表征。此后，将培养的细胞降至4％胎牛血清条件，并在存在或不存在T(3)(10(-8)M)的情况下培养随后的72小时。 Matrigel 侵袭测定表明，T3 处理显着增加了随后 24、48 和 72 小时培养的 EVT 的细胞投射数量。 RT-PCR分析显示，与对照培养物相比，在随后的24小时培养的EVT中，T(3)处理增加了MMP-2、MMP-3、胎儿FN和整合素α(5)β(1) mRNA的表达。免疫细胞化学和Western免疫印迹分析表明，与对照培养物相比，T(3)处理在随后的48小时培养的EVT中增加了MMP-2和MMP-3的表达。目前的结果表明，T(3) (10(-8) M) 可能在上调 EVT 侵入蜕膜的潜力中发挥重要作用。
It is well known that T(3) plays a crucial role in the maintenance of early pregnancy through the induction of endocrine function in villous trophoblasts. The effects of T(3) on extravillous trophoblast (EVT) function, however, remain to be elucidated. To investigate the possible role of T(3) in the regulation of EVT invasion to the decidua, we have examined whether T(3) affects EVT invasive potential and the expression of matrix metalloproteinase-2 (MMP-2), MMP-3, tissue inhibitor metalloproteinase-1, fetal fibronectin (FN), and integrin alpha(5)beta(1) in cultured early placental EVTs. Isolation and purification of trophoblasts differentiating into EVTs were performed by the enzymatic digestion of the anchoring chorionic villi, with the use of human FN-precoated culture dishes and FN-precoated Matrigel Transwells. The cells attached to the dishes were subcultured in DMEM supplemented with 10% fetal bovine serum for 48 h and were characterized by RT-PCR analysis after 24-h subculture and immunocytochemical analysis after 48-h subculture for specific EVT markers. Thereafter, the cultured cells were stepped down to a 4% fetal bovine serum condition and cultured in the presence or absence of T(3) (10(-8) M) for the subsequent 72 h. Matrigel invasion assay demonstrated that the treatment with T3 significantly increased the number of cell projections of subsequent 24-, 48-, and 72-h cultured EVTs. RT-PCR analysis revealed that the treatment with T(3) increased the expression of MMP-2, MMP-3, fetal FN, and integrin alpha(5)beta(1) mRNA in subsequent 24- h cultured EVTs compared with those in control cultures. Immunocytochemical and Western immunoblot analyses revealed that treatment with T(3) increased the expression of MMP-2 and MMP-3 in subsequent 48- h cultured EVTs compared with those in control cultures. The present results suggest that T(3) (10(-8) M) may play a vital role in up-regulating the invasive potential of EVTs into the decidua.