Protein glycosylation is a common post-translation modification and plays important roles in regulation of protein function. Especially, the abnormal glycosylation plays a critical role in the development and progression of tumor and the metastasis of cancer cells. MiRNAs also play a key role in the development and progression of cancer, but the mechanism how miRNAs affect on the biological functions and induce tumor malignant transformation via glycosylation needs further illustrated. In this study, the expression of glycosyltransferases were analyzed using glycan-related gene chip in miR-10b-overexpressed human normal mammary epithelial cells MCF10A. Then, the alteration of N-glycan and O-glycan was analyzed using the methods established in our laboratory. The expression of glycosyltransferase gene Fut8, MGAT3 and OGT and corresponding glycans were evaluated by qRT-PCR, Western blotting and lectin blotting. Taken together, our results provide the theoretical basis of glycomics on the role of miR-10b in breast cancer.