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  

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

作　　者：黄坷坷 齐博[1,2] 谷城威 霍书华 刘玉珍[1,2,3] 赵宝生 HUANG Ke-ke;QI Bo;GU Cheng-wei;HUO Shu-hua;LIU Yu-zhen;ZHAO Bao-sheng(Department of Thoracic Surgery,the First Affiliated Hospital of Xinxiang Medical University,He’nan Weihui 453100,China;Esophageal Cancer Institute of Xinxiang Medical University,He’nan Weihui 453100,China;Henan Neurology Institute at the First Affiliated Hospital of Xinxiang Medical University,He’nan Weihui 453100,China)

机构地区：[1]新乡医学院第一附属医院胸外科,河南卫辉453100 [2]新乡医学院食管癌研究所,河南卫辉453100 [3]河南省神经病学研究所,河南卫辉453100

出　　处：《解剖学报》2021年第5期737-743,共7页Acta Anatomica Sinica

基　　金：河南省重点科技攻关项目(202102310103);新乡市重点科技攻关项目(ZG15018);新乡医学院第一附属医院青年基金(QN-2019-A03)。

摘　　要：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sh NC和sh S100A6,Real-time PCR检测S100A6 m RNA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(cyclin D1)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sh S100A6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sh S100A6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,cyclin D1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。Objective To explore the effect of S100 calcium ion binding protein A6(S100 A6)on proliferation and migration of esophageal adenocarcinoma SK-GT-4 cells.Methods Lentiviruses were used to construct stable transfected cell lines(sh NC and sh S100 A6).Real-time PCR was used to detect the m RNA expression of S100 A6.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 S100 A6,p-ERK,p-Akt and its downstream molecular involved in proliferation and migration.Using U0126(inhibitor of MER1/2)and LY294002(inhibitor of PI3 K)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 sh S100 A6 cells.Results Stable cell lines of knockdown S100 A6 were constructed.Knockdown S100 A6 promoted cell proliferation and migration.Western blotting result displayed that in sh S100 A6 cells,the levels of p-Akt and p-ERK increased,p21 decreased,cyclin D1 increased,and the expression of β-catenin and vimentin,increased.U0126 and LY294002 inhibited the migration of sh S100 A6 cells.U0126 had no effect on the proliferation of sh S100 A6 cells,however LY294002 could inhibit the proliferation of sh S100 A6 cells.U0126 treatment on sh S100 A6 cells could decrease p-ERK and β-catenin expression.After sh S100 A6 cells treated with LY294002,p-Akt and β-catenin expression decreased,p21 expression increased and the expression of cyclin D1 decreased.Conclusion Low expression of S100 A6 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.

关 键 词：食管腺癌 S100钙离子结合蛋白A6 增殖 迁移 免疫印迹法 人 

分 类 号：R735.1[医药卫生—肿瘤]