Department of Obstetrics and Gynecology, University of British Columbia, Vancouver, British Columbia, Canada V6H 3V5.
Endocrinology. 2012 Feb;153(2):925-36. doi: 10.1210/en.2011-1488. Epub 2011 Dec 13.
The invasion of extravillous cytotrophoblasts (EVT) into the underlying maternal tissues and vasculature is a key step in human placentation. The molecular mechanisms involved in the development of the invasive phenotype of EVT include many that were first discovered for their role in cancer cell metastasis. Previous studies have demonstrated that N-cadherin and its regulatory transcription factor Twist play important roles in the onset and progression of cancers, but their roles in human trophoblastic cell invasion is not clear. The goal of the study was to examine the role of Twist and N-cadherin in human trophoblastic cell invasion. Twist and N-cadherin mRNA and protein levels were determined by RT-PCR and Western blotting in human placental tissues, highly invasive EVT, and poorly invasive JEG-3 and BeWo cells. Whether IL-1β and TGF-β1 regulate Twist mRNA and protein levels in the EVT was also examined. A small interfering RNA strategy was employed to determine the role of Twist and N-cadherin in HTR-8/SVneo cell invasion. Matrigel assays were used to assess cell invasion. Twist and N-cadherin were highly expressed in EVT but were poorly expressed in JEG-3 and BeWo cells. IL-1β and TGF-β1 differentially regulated Twist expression in EVT in a time- and concentration-dependent manner. Small interfering RNA specific for Twist decreased N-cadherin and reduced invasion of HTR-8/SVneo cells. Similarly, a reduction in N-cadherin decreased the invasive capacity of HTR-8/SVneo cells. Twist is an upstream regulator of N-cadherin-mediated invasion of human trophoblastic cells.
滋养细胞外突（EVT）侵入母体组织和血管是人类胎盘形成的关键步骤。参与 EVT 侵袭表型形成的分子机制包括许多最初在癌细胞转移中发现的机制。先前的研究表明，N-钙黏蛋白及其调节转录因子 Twist 在癌症的发生和进展中发挥重要作用，但它们在人滋养细胞侵袭中的作用尚不清楚。本研究旨在探讨 Twist 和 N-钙黏蛋白在人滋养细胞侵袭中的作用。采用 RT-PCR 和 Western blot 法检测人胎盘组织、高侵袭性 EVT 及低侵袭性 JEG-3 和 BeWo 细胞中 Twist 和 N-钙黏蛋白 mRNA 和蛋白的表达。还检测了 IL-1β 和 TGF-β1 是否调节 EVT 中 Twist mRNA 和蛋白的水平。采用小干扰 RNA 策略确定 Twist 和 N-钙黏蛋白在 HTR-8/SVneo 细胞侵袭中的作用。采用 Matrigel 测定法评估细胞侵袭。Twist 和 N-钙黏蛋白在 EVT 中高表达，但在 JEG-3 和 BeWo 细胞中低表达。IL-1β 和 TGF-β1 以时间和浓度依赖的方式差异调节 EVT 中 Twist 的表达。针对 Twist 的小干扰 RNA 降低了 N-钙黏蛋白的表达，并减少了 HTR-8/SVneo 细胞的侵袭。同样，N-钙黏蛋白的减少降低了 HTR-8/SVneo 细胞的侵袭能力。Twist 是 N-钙黏蛋白介导的人滋养细胞侵袭的上游调节因子。
