FUT3催化GLG1上Lea修饰促进胃癌细胞迁移侵袭的分子机制研究

Lewis抗原在胃癌进展中发挥着重要作用，FUT3是Lewis抗原合成过程中的关键酶，但其促进胃癌进展的机制尚不明确。前期研究结果表明，FUT3通过Lea糖修饰参与了胃癌细胞迁移侵袭过程。Lea免疫沉淀膜蛋白结合质谱及生信分析提示GLG1可能为FUT3作用的下游糖蛋白，提出“FUT3催化GLG1上Lea修饰促进胃癌细胞迁移侵袭”的假说。本研究拟收集胃癌组织样本分析FUT3、Lea、GLG1及其下游通路关键分子表达的相关性及临床意义；应用RNAi、Craspr-cas9技术、免疫沉淀、LC-MS/MS质谱分析、糖链解析、糖基化位点突变技术解析GLG1上Lea糖基化位点；通过免疫荧光、分子生物学及细胞功能实验明确FUT3催化GLG1上Lea修饰促进胃癌细胞迁移侵袭的分子机制；通过荷瘤动物模型进行在体验证。研究结果将为明确FUT3调控胃癌进展的分子机制提供新思路。

Lewis antigen plays an important role in the progression of gastric cancer（GC）,FUT3 is a key enzyme in the synthesis of Lewis antigen, but the molecular mechanism of its promotion of GC progression remains unclear.Our preliminary experimental results showed that FUT3 participated in the migration and invasion process of GC cells through Lea antigen expression.LC-MS/MS identification of Anti-Lea antiboty immunoprecipitated membrane proteins suggested that GLG1 might be the downstream glycoprotein of FUT3.Then we assume that FUT3 catalyzed Lea modification on GLG1 to promote the migration and invasion of GC cells.In this study, GC tissue samples were collected to analyze the correlation and clinical significance of FUT3,Lea,GLG1 and key molecules of downstream pathway expressions.The Lea glycosylation sites on GLG1 were analyzed by RNAi, CRASPR-Cas9, IP, LC-MS/MS,glycosylation chain analysis, and glycosylation site mutation techniques.IF,molecular biology and cell function experiments were conducted to clarify the molecular mechanism of FUT3 promoting the migration and invasion of GC cells by regulating Lea glycosylation on GLG1.Tumor bearing animal model were sestablished to validation the hypothesis in vivo.The results of this study will provide a new idea for clarifying the molecular mechanism of FUT3 regulating GC progression.