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To investigate miRNAs involved in the resistance of tongue cancer cells to chemotherapy, the miRNAs profiles between PYM (pingyangmycin) induced multidrug-resistant tongue cancer cell line Tca8113/PYM and its parent cell line Tca8113 were compared and miR-491-3p was identified to be downregulated in Tca8113/PYM cells.Functionally, restored miR-491-3p expression with mimics transfection sensitized Tca8113/PYM cells to PYM and cDDP, but miR-491-3p inhibitor transfection enhanced the resistance of Tca8113, SCC-25 and CAL-27 tongue cancer cell lines to PYM and cDDP.Mechanically, bioinformatics analysis combined with experimental validation demonstrated miR-491-3p directly targeted mTORC2 component Rictor.