Objective To investigate the underlying mechanism of miR-200c reversing the drug resistance of lung cancer cells to cisplatin (DDP). Methods After transfection of miR-200c mimics into lung cancer A549 cells, the expression of miR-200c was detected by real-time PCR. MTS assay was employed to measure the cell viability to observe the tolerance of A549 cells to DDP. Flow cytometry was used to measure the apoptosis rate of the cells, and Western blotting was applied to test the expression of survivin and Bcl-2 protein. After bioinformatics analysis showed that TUBB3, the downstream target molecule of miR-200c, was expressed differentially, luciferase reporter vectors containing wild type and mutant TUBB3 3′UTR were constructed, and the regulation effect and binding site of miR-200c to TUBB3 was studied by double luciferase reporter assay. The regulatory effect of miR-200c on the expression of TUBB3 protein, and on the expression of survivin and Bcl-2 protein were detected by Western blotting. SPSS statistics 19.0 was used to perform the statistical analysis. Results Compared with the control group, miR-200c transfection significantly increased the sensitivity of lung cancer cells to cisplatin [IC₅₀:(37.3±3.1) vs (15.3±3.3)μmol/L, P<0.01]. And the overexpression of miR-200c also resulted in the increased apoptosis rate of expression of A549/DDP cells (P<0.01), and decreased expression of survivin and Bcl-2 protein. The expression of TUBB3 at protein and mRNA levels was down-regulated in A549/DDP cells transfected with miR-200c mimics (P<0.01). Double luciferase reporter assay showed that miR-200c could regulate the expression of TUBB3 (P<0.01). All these results confirmed that TUBB3 was the downstream target gene of miR-200c. After pcDNA-TUBB3 plasmid was transfected, the expression of survivin and Bcl-2 protein in A549/DDP cells with miR-200c overexpression was up-regulated than that in the cells transfected with miR-200c mimics. Conclusion miR-200c can reverse the drug resistance of DDP-resistant lung cancer cells by down-regulating TUBB3 expression.