Cisplatin, quercetin and metformin were purchased from Sigma Biological Company (USA). Based on the reported plasma peak concentration (PPC) and pre-experiment results, the concentration gradient of cisplatin, metformin, and quercetin was determined as follows: cisplatin14 was 0, 0.25, 0.5, 1, 2, 4, 6, 8, 10, 12 μg/mL, metformin15 was 0, 0.25, 0.5, 1, 2, 4, 6, 8, 10, 12 mmol/L, and quercetin16 was 0, 10, 20, 30, 40, 50, 60, 70, 80, 90 μmol/L. When the drugs was applicated in combination, the low, medium and high concentrations were set according to the IC50 values of cisplatin, metformin and quercetin, which were expressed as: CDDP-L, CDDP-M, CDDP-H, Met-L, Met M, Met-H, Qu-L, Qu-M, Qu-H.
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