低表达S100钙离子结合蛋白A6促进食管腺癌SK-GT-4细胞的增殖和迁移

目的 探讨S100钙离子结合蛋白A6（S100A6）对食管腺癌SK-GT-4细胞增殖和迁移的影响。 方法 慢病毒转染，构建稳定细胞系shNC和shS100A6，Real-time PCR检测S100A6 mRNA表达情况；倒置显微镜和MTT检测细胞的增殖能力，Transwell检测细胞的迁移能力及U0126（MER1/2的抑制剂）、LY294002（PI3K抑制剂）对细胞增殖、迁移的影响；Western blotting检测细胞中S100A6、p-ERK、p-Akt及其下游参与增殖、迁移的相关蛋白的表达情况，检测U0126、LY294002对p-ERK、p-Akt及其下游参与增殖、迁移的相关蛋白表达的影响。 结果 构建了敲低S100A6的稳定细胞系，敲低S100A6促进了细胞的增殖和迁移，p-ERK、p-Akt水平升高，细胞周期抑制蛋白p21表达量下降、细胞周期蛋白D1(cyclinD1）表达升高，间质细胞标志蛋白——波形蛋白（vimentin）、β-连环蛋白（β-catenin）表达升高；U0126处理shS100A6细胞后，对细胞的增殖无影响，抑制细胞的迁移，p-ERK、β-catenin表达下降；LY294002处理shS100A6细胞后，抑制细胞的增殖和迁移，p-Akt表达下降，p21表达量升高，cyclinD1表达量下降，β-catenin表达下降。 结论 低表达S100A6促进细胞的增殖和迁移，其可能机制是通过p-Akt调控细胞周期的进程促进细胞增殖，通过激活p-Akt/p-ERK调控β-catenin促进细胞的迁移。

Low expression S100 calcium ion binding protein A6 promoting the proliferation and migration of esophageal adenocarcinoma SK-GT-4 cells

Objective To explore the effect of S100 calcium ion binding protein A6 (S100A6) on proliferation and migration of esophageal adenocarcinoma SK-GT-4 cells. Methods Lentiviruses were used to construct stable transfected cell lines (shNC and shS100A6). Real-time PCR was used to detect the mRNA expression of S100A6. The inverted microscope and MTT were used to detect cell proliferation. The Transwell assay was used to detect cell migration. Western blotting was used to detect the expression of S100A6, p-ERK, p-Akt and its downstream molecular involved in proliferation and migration. Using U0126 (inhibitor of MER1/2) and LY294002 (inhibitor of PI3K) to detect the effect of these two inhibitors on cell proliferation and migration and the expression of p-ERK, p-Akt and its downstream molecular involved in proliferation and migration in shS100A6 cells. Results Stable cell lines of knockdown S100A6 were constructed. Knockdown S100A6 promoted cell proliferation and migration. Western blotting result displayed that in shS100A6 cells, the levels of p-Akt and p-ERK increased, p21 decreased, cyclinD1 increased, and the expression of β-catenin and vimentin, increased. U0126 and LY294002 inhibited the migration of shS100A6 cells. U0126 had no effect on the proliferation of shS100A6 cells, however LY294002 could inhibit the proliferation of shS100A6 cells. U0126 treatment on shS100A6 cells could decrease p-ERK and β-catenin expression. After shS100A6 cells treated with LY294002, p-Akt and β-catenin expression decreased, p21 expression increased and the expression of cyclinD1 decreased. Conclusion Low expression of S100A6 promotes cell proliferation and migration, which may be mediated by activation of p-Akt regulating cell cycle progression to promote cell proliferation and by activation of p-Akt/p-ERK to regulate β-catenin to promote cell migration.