EpCAM associates with integrin and regulates cell adhesion in cancer cells
EpCAM associates with integrin and regulates cell adhesion in cancer cells
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DOI：
10.1016/j.bbrc.2019.11.152
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发表时间：
2020-02-19
影响因子：
3.1
通讯作者：
Gu, Jianguo
中科院分区：
文献类型：
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作者：
Yang, Jie;Isaji, Tomoya;Gu, Jianguo
上皮细胞粘附分子（EpCAM）是肿瘤相关抗原中表达频率最高、强度最强的抗原之一，但EpCAM在肿瘤细胞增殖、粘附和迁移中的作用尚不清楚。在本研究中，我们筛选了几种肿瘤细胞系，发现大肠癌CW-2和表皮样癌A431细胞表达相对较高水平的EpCAM。为了评估EpCAM表达在细胞粘附和迁移中的生物学功能，我们通过CRISPR/Cas9系统在这两种类型的癌细胞中建立了EpCAM基因的敲除（KO）。在EpCAM-KO细胞中，细长的细胞形态转化为圆形形态。这些细胞表现出细胞增殖和向细胞外基质蛋白迁移的减少，以及细胞信号传导元件如磷酸化粘着斑激酶（FAK）、AKT和ERK的减少。此外，在EpCAM-KO细胞中，细胞生长和殖民地形成能力显著降低。重要的是，免疫共沉淀分析显示EpCAM与整合素β 1相关。此外，与野生型细胞相比，EpCAM-KO细胞中整合素α 5的表达水平降低。综上所述，这些数据清楚地表明EpCAM与整合素β 1结合以调节FAK/ERK信号通路，通过细胞外基质粘附来控制细胞粘附、迁移和增殖，这为EpCAM介导的生物学功能和癌症表型提供了新的机制。(C)2019爱思唯尔公司All rights reserved.
The epithelial cell adhesion molecule (EpCAM) is one of the most frequently and intensely expressed of tumor-associated antigens, but the role that EpCAM plays in the proliferation, adhesion and migration properties of cancer cells remains unclear. In the present study, we screened several tumor cell lines and found that colorectal cancer CW-2 and epidermoid carcinoma A431 cells expressed relatively higher levels of EpCAM. In order to assess the biological functions of EpCAM expression in cell adhesion and migration, we established a knock out (KO) of EpCAM genes in both of these types of cancer cells via a CRISPR/Cas9 system. The elongated cell morphology was converted to a rounded morphology in the EpCAM-KO cells. These cells showed decreases in cell proliferation and migration into extracellular matrix proteins, as well as decreases in cellular signaling elements such as phosphorylated focal adhesion kinase (FAK), AKT and ERK. Moreover, the cell growth and the colony formation abilities were significantly decreased in EpCAM-KO cells. Importantly, co-immunoprecipitation analysis revealed that EpCAM associated with integrin beta 1. Also, the expression levels of integrin alpha 5 were decreased in EpCAM-KO cells, compared with that in the wild-type cells. Taken together, these data clearly demonstrate that EpCAM associates with integrin beta 1 to regulate FAK/ERK signaling pathways in controlling cell adhesion, migration and proliferation via extracellular matrix adhesion, which provides novel mechanisms for EpCAM-mediated biological functions and cancer phenotypes. (C) 2019 Elsevier Inc. All rights reserved.