[目的]探讨MUC16蛋白的硫酸化O型连接糖对食管鳞癌细胞的迁移作用.[方法]TCGA数据库分析MUC16在食管癌组织中的表达水平.过表达或敲低MUC16后,划痕实验检测食管鳞癌细胞KYSE-30的迁移水平.过表达或敲低CHST4后,Lectin blot和Alcian blue染色检测食管鳞癌细胞中MUC16的O型连接糖硫酸化水平以及食管鳞癌细胞的迁移水平,LC-ESI-MS检测MUC16的O型连接糖类型.[结果]相比于食管正常组织,食管癌组织中MUC16和 CHST4 的表达水平较高(0.78±0.13 vs 0.09±0.02;0.43±0.08 vs 0.07±0.02,P＜0.05).过表达 MUC16 后,食管鳞癌细胞的迁移水平显著上升;敲低MUC16后,食管鳞癌细胞的迁移水平显著下降.敲低CHST4后,MUC16 O型连接糖的硫酸化水平显著下降,食管鳞癌细胞的迁移水平显著下降;过表达CHST4后,MUC16 O型连接糖的硫酸化水平显著上升,食管鳞癌细胞的迁移水平显著上升.食管鳞癌细胞在m/z 813和1121处发现了含有GlcNAc-6-O-硫酸的低聚糖,而敲低CHST4后硫酸化的低聚糖水平降低.[结论]CHST4介导的MUC16蛋白O型连接糖硫酸化对食管鳞癌细胞的迁移具有促进作用(1.00±0.05 vs 1.83±0.12).
Promotion of migration of esophageal squamous carcinoma cells by sulfated O-linked glycans of MUC16
[Objective]To investigate the migratory effect of sulfated O-linked glycans of MUC16 protein on esophageal squa-mous carcinoma cells.[Method]TCGA database was analyzed for the expression level of MUC16 in esophageal cancer tissues.After overexpression or knockdown of MUC16,the migration level of KYSE-30 in esophageal squamous carcinoma cells was detected by scratch assay.After overexpression or knockdown of CHST4,Lectin blot and Alcian blue staining were performed to detect the sulfation level of O-linked glycans of MUC16 protein in esophageal squamous carcinoma cells and the migration lev-el of esophageal squamous carcinoma cells,and LC-ESI-MS was performed to detect the O-linked glycan type of MUC16 protein.[Result]Compared with normal esophageal tissues,the expression level of MUC16 and CHST4 in esophageal cancer tissues was higher(0.78±0.13 vs 0.09±0.02,P＜0.05)and the expression level of CHST4 was higher(0.43±0.08 vs 0.07±0.02,P＜0.05).After overexpression of MUC16,the migration level of esophageal squamous cell cells increased significant-ly.After MUC16 was knocked down,the migration level of esophageal squamous cell cells decreased significantly.After CHST4 was knocked down,the sulfation level of MUC16 O-type linked sugar was significantly decreased,and the migration level of e-sophageal squamous cell carcinoma was significantly decreased.After overexpression of CHST4,the sulfation level of MUC16 O-type link-sugar was significantly increased,and the migration level of esophageal squamous cell carcinoma was significantly increased.GlcNAc-6-O-sulfuric acid oligosaccharides were found in esophageal squamous cell cells at m/z 813 and 1121,while sulfated oligosaccharides were reduced after CHST4 knockdown.[Conclusion]Chst4-mediated MUC16 protein O-type link-sugar sulfation promoted the migration of esophageal squamous cell cells(1.00±0.05 vs 1.83±0.12).
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