In vivo gene manipulations of epithelial cell sheets: a novel model to study epithelial-to-mesenchymal transition.
In vivo gene manipulations of epithelial cell sheets: a novel model to study epithelial-to-mesenchymal transition.
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上皮细胞片的体内基因操作：研究上皮间质转化的新模型。
DOI：
10.1111/j.1440-169x.2011.01252.x
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Y.
中科院分区：
文献类型：
--
作者：
Yoshino;T.;Saito;D.;Tadokoro;R. and Takahashi;Y.
胚胎细胞按其形态可分为两类：上皮细胞和间充质细胞。在发育过程中的动态形态发生过程中，上皮细胞通常通过上皮向间充质转化(EMT)的过程向间充质转化。EMT是肿瘤转移过程中的一个中心问题，在肿瘤转移过程中，上皮来源的肿瘤细胞转化为具有高迁移性的间质细胞。虽然许多分子已被确定参与EMT，主要是通过体外研究，但在活体模型系统中一直有限。我们在这里建立了一种新的模型，利用该模型可以直接在活体内分析EMT。通过电穿孔技术，我们瞄准了侧板中胚层的一部分，它形成了描绘肾脏区域的上皮细胞片，称为肾体腔上皮(NEPH-CE)。增强的绿色荧光蛋白-电穿孔的NEPH-CE保持了上皮的完整性，而不会侵犯下面的间质(中肾)。NEPH-CE转基因使我们进一步探索体内EMT的诱导物，发现Ras-Raf和RhoA信号是有效的诱导物。实时成像共聚焦显微镜显示，在EMT过程中，细胞开始向基质延伸细胞突起，随后它们的细胞体移位。此外，我们建立了对EMT诱导的细胞的长期跟踪，这些细胞动态地重新定位在肾间质中。NEPH-CE转基因将为直接在实际体内研究EMT的细胞和分子机制开辟了一条途径。
Embryonic cells are classified into two types of cells by their morphology, epithelial and mesenchymal cells. During dynamic morphogenesis in development, epithelial cells often switch to mesenchymal by the process known as epithelial‐to‐mesenchymal transition (EMT). EMT is a central issue in cancer metastasis where epithelial‐derived tumor cells are converted to mesenchymal with high mobility. Although many molecules have been identified to be involved in the EMT mostly byin vitrostudies,in vivomodel systems have been limited. We here established a novel model with which EMT can be analyzed directly in the living body. By an electroporation technique, we targeted a portion of the lateral plate mesoderm that forms epithelial cell sheets delineating the kidney region, called nephric coelomic epithelium (Neph‐CE). Enhanced green fluorescent protein‐electroporated Neph‐CE retained the epithelial integrity without invading into the underling stroma (mesonephros). The Neph‐CE transgenesis further allowed us to explore EMT inducersin vivo, and to find that Ras‐Raf and RhoA signals were potent inducers. Live‐imaging confocal microscopy revealed that during EMT processes cells started extending cellular protrusions toward the stroma, followed by translocation of their cell bodies. Furthermore, we established a long‐term tracing of EMT‐induced cells, which were dynamically relocated within the kidney stroma. The Neph‐CE‐transgenesis will open a way to study cellular and molecular mechanisms underlying EMT directly in actual body.