Y of MMP2 and MMP9 and discovered that the expression and activity were inhibited by the treatment of shikonin in a dosedependent manner in both cell lines. As described above, shikonin inhibited the migration and invasion of U87 and U251 cells. Offered these benefits, it’s affordable to infer that shikonin exhibited inhibitory effects on glioma cell migration and invasion by inhibiting the expression and activity of MMP2 and MMP9. It has been reported that the Wntcatenin pathway plays a vital part inside the tumorigenesis and progression of gliomas [36]. catenin is upregulated and accumulated in the cytoplasm and nucleus of glioma cells in comparison to the typical brain tissue along with the expression levels are positively correlated with histological malignancy [37,38]. Accumulated catenin also promotes the proliferation, migration, and invasion of glioma cells [39,40]. Additionally, knockdown of catenin inhibits cell proliferation and induces apoptotic cell death in U251 cells [24]. Deregulation of catenin correlates to the tumorigenesis of gliomas. Since the 1st step in catenin degradation is phosphorylation by CK1 around the multiprotein destruction complex [25], we investigated the expression levels of pcatenin with all the treatment of shikonin. U87 and U251 cells have been Iron sucrose Protocol treated with shikonin in the concentrations of 2.5, five, and 7.5 molL for 48 h. Interestingly, the outcomes showed contrary trends in the two cell lines. The expression of pcatenin Y333 was significantly inhibited in U87 cells but upregulated in U251 cells. Nevertheless, pcatenin Ser45 expression was not changed by shikonin, which means that it was not involved in the process. The contrary trend in pcatenin Y333 expression may well be as a result of the following cause. The two cell lines employed within this study have different p53 status. U87 cells are p53 wildtype glioblastoma cells while U251 cells are p53 mutant [41,42]. It was previously reported that catenin expression was regulated by the p53 pathway: upregulated or accumulated p53 lowered catenin expression [43]. Nonetheless, the downregulation of catenin could only be obtained by wildtype p53 as opposed to cancerassociated mutant variety p53 [43]. However, p53 mutation could induce aberrant accumulation of catenin in different human cancers [44]. This may well clarify why catenin expression was constant in p53 mutant U251 cells. Therapy of shikonin possibly induced downregulation of catenin Y333 in p53 wildtype U87 cells. Nevertheless in U251 cells, catenin could not be regulated by mutant p53 as well as the downstream ubiquitinproteasomedependent degradation of catenin was activated, which promoted pcatenin Y333 expression in U251 cells. In this operate, we confirmed that pcatenin Y333 knockdown or overexpression could attenuate or improve the migration, invasion, MMP expression and activity in U87 cells. MMP2 and MMP9 are the downstream targets of catenin [45,46]. Meanwhile, it has been established that catenin knockdown resulted in inhibited expression of MMP2 and MMP9 [47].Int. J. Mol. Sci. 2015,Downregulated catenin expression results in decreased expression of MMP2 and MMP9 [47,48]. Consequently, attenuated catenin expression also as downstream MMP2 and MMP9 inhibition could be among the possible mechanisms within the shikonininduced inhibition of glioma cell migration and invasion. Shikonin inhibited the proliferation, migration, and invasion in each cell lines by inhibiting MMP2, MMP9 and targeting pcatenin Y333. Shikonin possibly displayed inhibitory effects through differen.