Park Song-Yi, Jang Hwanseok, Kim Seon-Young, Kim Dasarang, Park Yongdoo, Kee Sun-Ho
Department of Microbiology, College of Medicine, Korea University, Seoul 02841, Korea.
Department of Biomedical Sciences, College of Medicine, Korea University, Seoul 02841, Korea.
Bioengineering (Basel). 2021 May 11;8(5):65. doi: 10.3390/bioengineering8050065.
Collective cell migration of epithelial tumor cells is one of the important factors for elucidating cancer metastasis and developing novel drugs for cancer treatment. Especially, new roles of E-cadherin in cancer migration and metastasis, beyond the epithelial-mesenchymal transition, have recently been unveiled. Here, we quantitatively examined cell motility using micropatterned free edge migration model with E-cadherin re-expressing EC96 cells derived from adenocarcinoma gastric (AGS) cell line. EC96 cells showed increased migration features such as the expansion of cell islands and straightforward movement compared to AGS cells. The function of tight junction proteins known to E-cadherin expression were evaluated for cell migration by knockdown using sh-RNA. Cell migration and straight movement of EC96 cells were reduced by knockdown of ZO-1 and claudin-7, to a lesser degree. Analysis of the migratory activity of boundary cells and inner cells shows that EC96 cell migration was primarily conducted by boundary cells, similar to leader cells in collective migration. Immunofluorescence analysis showed that tight junctions (TJs) of EC96 cells might play important roles in intracellular communication among boundary cells. ZO-1 is localized to the base of protruding lamellipodia and cell contact sites at the rear of cells, indicating that ZO-1 might be important for the interaction between traction and tensile forces. Overall, dynamic regulation of E-cadherin expression and localization by interaction with ZO-1 protein is one of the targets for elucidating the mechanism of collective migration of cancer metastasis.
上皮肿瘤细胞的集体迁移是阐明癌症转移和开发新型癌症治疗药物的重要因素之一。特别是，E-钙黏蛋白在癌症迁移和转移中的新作用，超出了上皮-间质转化，最近已被揭示。在这里，我们使用微图案化自由边缘迁移模型，对源自胃腺癌（AGS）细胞系的E-钙黏蛋白重新表达的EC96细胞进行了细胞运动性的定量检测。与AGS细胞相比，EC96细胞表现出增加的迁移特征，如细胞岛的扩展和直线运动。通过使用sh-RNA敲低来评估已知与E-钙黏蛋白表达相关的紧密连接蛋白对细胞迁移的功能。敲低ZO-1和claudin-7后，EC96细胞的迁移和直线运动有所减少，但程度较轻。对边界细胞和内部细胞迁移活性的分析表明，EC96细胞的迁移主要由边界细胞进行，类似于集体迁移中的引导细胞。免疫荧光分析表明，EC96细胞的紧密连接（TJs）可能在边界细胞之间的细胞内通讯中起重要作用。ZO-1定位于突出的片状伪足基部和细胞后部的细胞接触部位，表明ZO-1可能对牵引力和拉力之间的相互作用很重要。总体而言，通过与ZO-1蛋白相互作用对E-钙黏蛋白表达和定位进行动态调节是阐明癌症转移集体迁移机制的目标之一。
