功能蛋白O-GlcNAc 糖基化修饰调控绒毛膜癌细胞的转移.pdfVIP

中国细胞生物学学报 Chinese Journal of Cell Biology 2013, 35(7): 922–927 http ://www.cj DOI: 10.11844/cj cb.2013.07.0018 功能蛋白O-GlcNAc糖基化修饰调控 绒毛膜癌细胞的转移 屈茹楠 琚娜娜 吴明军 赵德璋 王应雄 杨 竹 于 超* (重庆医科大学生命科学研究院, 重庆400016) 摘要 为研究氧连接N- 乙酰葡萄糖胺(O-linked N-acetylglucosamine, O-GlcNAc)糖基化修 饰调控人绒毛膜癌细胞(JAR)迁移的分子机制, 首先采用siRNA和酶特异性抑制剂作用细胞, 构 建O-GlcNAc修饰总蛋白低表达和高表达的细胞模型; 利用Transwell方法及黏附实验比较细胞迁 β 移及与血管内皮细胞黏附能力的变化; 并通过免疫共沉淀技术检测重要的功能蛋白 -catenin 的 O-GlcNAc程度, 从而分析蛋白转录翻译后调控对肿瘤细胞迁移影响的分子机制。结果表明, 增加 JAR细胞中O-GlcNAc修饰水平可促进细胞迁移及与血管内皮细胞的黏附活性, 且O-GlcNAc修饰作 β 用的靶蛋白 -catenin 的糖基化水平也显著增加。当下调细胞中O-GlcNAc蛋白修饰水平, 其迁移及 黏附能力随之下降。表明O-GlcNAc修饰参与了绒毛膜癌细胞转移的调控。 关键词 O-GlcNAc; 绒毛膜癌; 细胞迁移; 黏附; β-catenin O-GlcNAcylation of Functional Protein Regulates Human Choriocarcinoma Cells Migration Qu Runan, Ju Nana, Wu Mingjun, Zhao Dezhang, Wang Yingxiong, Yang Zhu, Yu Chao* (Institute of Life Science, Chongqing Medical University, Chongqing 400016, China) Abstract To investigate the molecular mechanisms involved in human choriocarcinoma cells (JAR) migration regulated by O-GlcNAc glycosylation, cell models expressing different O-GlcNAcylation levels were established by siRNA interference or OGA inhibitor. Transwell assay and adhesion assay were used to examine JAR cell migration ability and adhesion to EA.hy926 cells. Meanwhile, the O-GlcNAcylation level of β-catenin was detected by immunoprecipitation. The results indicated O-GlcNAcylation enhances the JAR cell migration and adhesion to endothelial cells in vitro. Moreover, the O-GlcNAcylation of β-catenin but not the protenin level increased apparently. The mig