Yes-associated protein (YAP) regulates numerous cellular homeostasis processes and malignant transformation. We found that YAP influences ZO-1-mediated cell migration using E-cadherin-restored EC96 cells derived from gastric malignant AGS cells. Ectopic expression of E-cadherin enhanced straightforward migration of cells, in comparison to the meandering movement of parental AGS cells. In EC96 cells, YAP and ZO-1 expression increased but nuclear YAP levels and activity were reduced. Nuclear factor-κB (NF-κB) mediated the increase in ZO-1 expression, possibly stabilizing cytoplasmic YAP post-translationally. Downregulation of YAP expression using siYAP RNA or stable knock-down inhibited straightforward cell migration by fragmenting ZO-1 containing tight junctions (TJs) but not adherens junctions, implying involvement of YAP in ZO-1-mediated cell migration. The association of YAP with ZO-1 was mediated by angiomotin (AMOT) because downregulation of AMOT dissociated YAP from ZO-1 and reduced cell migration. E-cadherin restoration in malignant cancer cells induced NF-κB signaling to enhance ZO-1 expression and subsequently stabilize YAP. At high expression levels, YAP associates with ZO-1 via AMOT at TJs, influencing ZO-1-mediated cell migration and maintaining TJ integrity.

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.

Objective: The purpose of this study was to evaluate the relations among the combined therapy with steroid and the detoxification enzyme gene polymorphisms in patients with sudden sensorineural hearing loss (SSNHL). The pathogenetic mechanism of inner ear dysfunction could involve an increase in lipid peroxidation and a decrease in cellular antioxidant defense. Glutathione S-transferases (GSTs) and cytochrome P450 (CYP) belong to a system of detoxification and antioxidant enzymes that have been demonstrated in the inner ear. Study Design: A prospective study in patients with SSNHL. Patients and Methods: All 441 subjects were genotyped for GSTM1, GSTT1, and CYP1A1 polymorphisms. The polymorphisms were analyzed by polymerase chain reaction amplification, restriction enzyme digestion, and deoxyribonucleic acid fragment separation by electrophoresis. Results: No significant difference was observed between SSNHL patients and controls in 3 polymorphisms. However, the prevalence of the partial recovery group in patients with the CC genotype of CYP1A1 (22%) was higher than that in the complete recovery (7.4%) or no recovery group (12.5%) for the subjects classified according to modified Siegel's criteria but were not statistically significant. Conclusion: This is the first approach to analyze gene polymorphism and efficacy of clinical treatment of patients with SSNHL, although the observations do not confirm the effect of the GSTM1/T1 and CYP1A1 genotypes as a risk factor for SSNHL.