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蛋白质糖基化是蛋白质翻译后修饰之一,对蛋白质功能有重要的调节作用,而异常糖基化在肿瘤的发生、发展以及癌细胞转移过程中起到关键作用.MiRNAs在癌症的发生发展过程中同样起到非常关键的作用,但其如何影响糖基化进而在肿瘤恶性转化过程中发挥生物学功能的研究甚少.本文将miR-10b在人正常乳腺上皮细胞MCF10A中过表达,利用糖类相关基因芯片系统筛选了发生显著变化的糖基转移酶;随后利用本实验室建立的N-糖链及O-糖链测定方法,分析糖链水平的表达差异;最后对关键糖基转移酶基因Fut8、MGAT3及OGT通过荧光定量PCR、蛋白质免疫印迹和凝集素免疫印迹进行了验证,为研究miR-10b在乳腺癌中的作用提供更多糖组学方面的理论基础.
Protein glycosylation is one of the posttranslational modifications of proteins, which plays an important regulatory role in protein function, and abnormal glycosylation plays a key role in tumorigenesis, development and metastasis of cancer cells.MiRNAs play an important role in the development of cancer Also plays a crucial role in the process of tumorigenesis.However, little is known about how miR-10b plays a role in the malignant transformation of tumor.This article overexpresses miR-10b in human normal breast epithelial cells MCF10A, Related gene microarray screening of glycosyltransferases that have undergone significant changes; then using the N-glycan and O-glycan assay methods established in our laboratory to analyze the expression differences of sugar chain levels; finally, the key glycosyltransferases Genes Fut8, MGAT3 and OGT were verified by real-time PCR, Western blotting and lectin immunoblotting, providing more theoretical basis for studying glycomics in order to study the role of miR-10b in breast cancer.