Polypeptide, N-acetylgalactosaminyltransferase 2 (ppGalNAc-T2), is a member of the ppGalNAcT family which is the initial glycosyltransferase in O-glycan synthesis. Our previous studies have demonstrated that ppGalNAc-T2 plays a vital role in the process of tumor emergence and development. In this study, we aimed to determine whether ppGalNAc-T2 is correlated with the invasion and metastasis of human glioma cells. PpGalNAc-T2 sense vectors and interference vectors constructed in our laboratory were successfully transfected into SHG44 and U251 cells, which were observed through fluorescence microscopy, RT-PCR and western blot analysis. The metastasis of U251 cells was determined using a wound healing assay, and ppGalNAc-T2 was found to inhibit cell migration. By RT-PCR, glioma cell invasion assay showed that the expression of MMP-2 was increased in ppGalNAc-T2-knockdown cells, while the expression of MMP-9 and TGF-β1 was decreased in ppGalNAc-T2-overexpressing cells at the mRNA level. Meanwhile, the expression of TIMP-2 at the mRNA level was contrary to MMP-2. In summary, our findings indicate that ppGalNAc-T2 plays a poten-tial role in glioma cell migration and invasion, suggesting a new molecular therapeutic target for human malignant glioma treatment.