Modulation of ezrin and E-cadherin expression by IL-1β and TGF-β1 in human trophoblasts
Modulation of ezrin and E-cadherin expression by IL-1β and TGF-β1 in human trophoblasts
复制标题
DOI：
10.1016/j.jri.2004.04.005
复制
发表时间：
2004-12-01
影响因子：
3.4
通讯作者：
Das, C
中科院分区：
文献类型：
--
作者：
Karmakar, S;Das, C
目的：本研究检测IL-1 β和TGF-β 1在ezrin调节中的作用。人滋养层细胞中的E-cadherin、CD 44和β-catenin表达可能导致它们在细胞与细胞和细胞与基质相互作用期间改变细胞骨架动力学。滋养层用IL-1 β或TGF-β 1刺激分离纯化的早期和足月胎盘细胞和人绒毛膜癌细胞系JEG-3，（10 ng/ml）孵育12 h，随后对埃兹蛋白、E-钙粘蛋白、CD 44和β-连环蛋白进行RT-PCR。这些蛋白质的免疫定位在绒毛膜绒毛以及在细胞因子刺激的培养细胞中进行。采用Western Blot方法研究ezrin和E-cadherin在原发性绒毛外膜中的表达调控。绒毛和绒毛滋养层细胞。结果：IL-1 β可下调滋养层细胞和JEG-3细胞ezrin、E-cadherin和β-catenin的表达，上调CD 44信使的表达。相反的，我不知道。TGF-β 1表现出相反的效果。即上调Ezrin。E-钙粘蛋白β-连环蛋白和下调CD 44。这些观察结果进一步证实了上述蛋白质在妊娠早期和足月绒毛组织中的免疫定位结果，前者以IL-1 β为主，后者以TGF-β 1为主[Am. J. Reprod. Immunol.48（2002）210]。通过SEM观察的细胞形态显示，IL-1 β刺激后细胞与基质的粘附增强，细胞与细胞的相互作用较差，TGF-β 1存在时细胞间粘附增强，细胞与基质的相互作用较弱。结晶紫染色和Matrigel侵袭显示IL-1 β刺激后侵袭指数较高，TGF-β 1刺激后侵袭指数较低。IL-1 β介导的细胞-基质相互作用增加，细胞-细胞粘附沿着降低，同时ezrin和E-钙粘蛋白表达降低，这与侵袭力增强有关，而TGF-β 1介导的细胞-细胞粘附上调，细胞-细胞粘附降低，基质相互作用沿着增加的埃兹蛋白和E-钙粘蛋白表达与侵袭性降低相关，沿着改变的细胞形态。因此，这些事实表明这两种细胞因子通过改变细胞-基质和细胞-细胞相互作用在细胞运动和侵袭过程中可能发挥作用。(C)2004爱思唯尔爱尔兰有限公司保留所有权利。
Objectives: The present study examines the effects of IL-1beta and TGF-beta1 in modulation of ezrin. E-cadherin, CD44 and beta-catenin expression in human trophoblast cells which may lead to their altered cytoskeleton dynamics during cell-to-cell and cell-to-matrix interactions.Methods: Trophoblast (extravillous and villous) cells isolated and purified front early and term placentae and human choriocarcinoma cell line JEG-3 used in this study were challenged with either IL-1beta or TGF-beta1 (10 ng/ml) for 12 h following which RT-PCR was performed for ezrin, E-cadherin, CD44 and beta-catenin. Immunolocalization of these proteins was carried out in the chorionic villi as well as in the cultured cells stimulated by the cytokines. Western Blot was performed to study the regulation of ezrin and E-cadherin in primary extravillous. villous and term trophoblast by these cytokines. Scanning electron microscopy (SEM) and Matrigel Invasion Assay was used to study the effect of these cytokines on cellular morphology and invasion.Results: IL-1beta induced a down regulation in the expression of ezrin, E-cadherin and beta-catenin while upregulation of CD44 message in both primary trophoblast and JEG-3 cells. On the contrary. TGF-beta1 exhibited just an opposite effect. i.e. up regulation of ezrin. E-cadherin, beta-catenin. and down regulation of CD44. These observations were further corroborated with the immunolocalization findings of the above proteins in first trimester and term villous tissue, the former having predominance of IL-1beta and the latter of TGF-beta1 [Am. J. Reprod. Immunol. 48 (2002) 210]. Cellular morphology as observed through SEM revealed ail enhanced cell-to-rnatrix adhesion with poor cell-cell interaction following IL-1beta challenge and a strong intercellular adhesion with weak cell-to-matrix interaction in presence of TGF-beta1. Crystal violet staining and Matrigel invasion revealed a higher invasion index following IL-1beta challenge and a low invasion index following TGF-beta1 challenge.Conclusion: IL-1beta mediated increased cell-to-matrix interaction with reduced cell-to-cell adhesion along with reduced ezrin and E-cadherin expression is associated with enhanced invasiveness while TGF-beta1 mediated up regulation of cell-to-cell adhesion with reduced cell-to-matrix interaction along with an increased ezrin and E-cadherin expression, is associated with reduced invasiveness, along with an altered cellular morphology. These facts therefore indicate the possible role of the two cytokines during cell motility and invasion through alteration of cell-matrix and cell-cell interaction. (C) 2004 Elsevier Ireland Ltd. All rights reserved.