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Glioblastoma multiforme (GBM) is among the most devastating types of cancer, with a median survival of <1 year. despite the development of new surgical and radiation techniques, and the use of multiple anti-neoplastic drugs, effective treatment strategies for malignant gliomas have not yet been developed. The limited efficacy of current treatments reflects the resistance of glioblastoma cells to cytotoxic agents. In this study, using western blot analysis, we found that Yin Yang 1 (YY1) expression was increased in cisplatin-resistant glioblastoma U87MG cells (U87MG-cR). We observed that the silencing of YY1 sensitized the U87MG-cR cells to cisplatin and that the overexpression of YY1 promoted the resistance of LN-229 glioblastoma cells to cisplatin, as shown by MTT assay. Using sphere formation assay, we also found that the silencing of YY1 inhibited the formation of the glioblastoma-initiating cell (GIc) phenotype in the U87MG-cR cells. In addition, the results of RT-qPcR revealed that miR-186 expression was decreased in U87MG-CR cells. Using RT-PCR and western blot analysis, we observed that overexpression of miR-186 inhibited YY1 expression in U87MG-CR cells. The overexpression of miR-186 also reversed cisplatin resistance and the formation of the GIc phenotype in glioblastoma cells. On the whole, the findings of this study demonstrate that miR-186 reverses cisplatin resistance and inhibits the formation of the GIc phenotype by degrading YY1 in glioblastoma.