CD147/CAS介导的EMT参与子宫内膜异位症发病机制的研究

Endometriosis is a common gynecological disease, which has become a common cause of female infertility. However, the involved mechanism remains exclusive. In our previous studies, we have demonstrated the high expression of CD147 in endometriosis lesions and its key roles in regulating the cell apoptosis and migration during the endometriosis progression. Our preliminary study found that CAS is a novel interacting protein of CD147. In human endometriotic tissues, CAS was down-regulated and negatively correlated with CD147 expression. Furthermore, CD147 could interact with CAS and E-cadherin and CAS was able to interact with CD147 and E-cadherin, suggesting that the three proteins were in a complex and involved in the EMT process of human endometrial epithelial cells during the endometriosis. In the present study, we will further confirm the expression pattern of CAS, E-cadherin and its downstream EMT related molecules and investigate their correlation among those molecules. We also propose using CD147/CAS overexpression or knockout cell models to test the interacting pattern of CD147, CAS and E-cadherin. EMT related signaling pathways will be also determined in these cell models. The findings of the proposed study would provide the possible molecular mechanisms under the regulation of CD147/CAS in the progress of endometriosis and may provide a potential target for diagnosis and treatment of endometriosis.

子宫内膜异位症（内异症）已成为导致女性不孕不育的主要疾病，然而其致病机制尚不清楚。我们的研究已揭示CD147在异位内膜中表达上调，它是调控子宫内膜上皮细胞凋亡和迁移的重要分子。我们前期研究发现一个与CD147相互作用的新蛋白CAS，高表达CD147的异位内膜中CAS的表达降低；CD147、CAS和E-cadherin形成的复合体参与了CD147介导的上皮-间充质转化（EMT）。上述结果提示，CD147与CAS的相互作用可能是调控子宫内膜上皮细胞EMT的关键因素。本研究将检测人异位内膜中CAS及EMT通路分子的表达，分析它们与CD147表达的相关性；利用基因敲除／过表达CD147/CAS的子宫内膜上皮细胞模型，研究CD147、CAS和E-cadherin的相互作用模式及其调节的EMT信号途径。研究结果将阐明CD147/CAS介导的EMT在内异症中的作用及分子机制，为内异症的诊治提供潜在靶点。

子宫内膜异位症（简称内异症）是一种常见妇科疾病，其中子宫内膜细胞的迁移以及恶性侵袭行为是内异症发生的关键步骤。有研究表明，上皮-间充质转化（Epithelial–mesenchymal transition, EMT）在内异症的发生发展过程中具有重要的作用。然而，对EMT过程是如何影响子宫内膜异位症的发生及其分子机制尚无深入的研究。我们前期研究发现CD147在子宫内膜异位症中的表达异常升高并通过CD147-erk-Bcl2信号通路增强子宫内膜上皮细胞的存活能力，但CD147调节子宫内膜上皮细胞的迁移并不依赖于CD147常见的下游调控分子-基质金属蛋白酶家族分子（MMPs）。本项目在前期工作的基础上，发现了一个新的CD147直接相互作用的蛋白- CAS，并阐明了CD147/CAS/E-cadherin复合体参与子宫内膜上皮细胞EMT过程的分子机制。我们的实验结果发现，过表达CD147可以促进子宫内膜上皮细胞的迁移以及EMT过程，而敲减CAS后，促进EMT的分子标志物基因Vimentin和下游分子Snail的表达也显著上调。过表达CAS后，细胞的迁移和侵袭被显著抑制，表明CAS具有负调控EMT的作用。免疫共沉淀结果发现CD147、CAS和E-cadherin形成了三聚复合体，过表达CD147可以破坏CAS / E-cadherin / β-catenin复合物的稳定性，进而导致激活的β-catenin入核，促进EMT过程。有趣的是，过表达CD147对CAS的表达并无影响，针对CAS启动子活性的双荧光素酶结果也进一步证明了CD147对CAS基因的转录没有调节作用。此外，在我们收集到的子宫内膜异位症患者的样本中也发现， CAS的表达显著降低并与CD147的高表达呈负相关；子宫异位内膜中上皮中的E-cadherin表达降低，而入核的β-catenin表达增加，表明CAS的下调促进了β-catenin信号的激活，进而调节子宫内膜上皮细胞的EMT过程。综上所述，我们的研究阐明了CD147 通过CAS介导的EMT过程在子宫内膜异位症中分子机制，CD147的异常高表达破坏了CD147/CAS/E-cadherin复合体的稳定结合，激活EMT相关的信号通路，介导了子宫内膜上皮细胞的EMT过程和细胞的迁移/侵袭，进而参与子宫内膜异位症的发生发展，为子宫内膜异位症的诊治提供了新的靶点。