Objective  To study the effect of hyperthermia on expressions of multidrug resistance（MDR）genes intongue squamous cell carcinoma cell line Tca8113 and its MDR cell line Tca8113/CBDEA and intracellular ADM concentration. Methods  Cell lines were hyperthermia-treated（42 ℃ for 0.5 h）, 4 h and 24 h later real-time quanti-tative reverse transcriptase-polymerase chain reaction（RT-PCR） detected expressions of MDR relative genes（MDR1, MRP1, GST-π）and HTS 7000 Plus Bio Assay Reader measured intracellular ADM concentration. Results  Expres-sions of MDR1, MRP1, GST-π genes in Tca8113/CBDEA significantly descended at 4 h and 24 h post-hyperthermia（P<0.01）. Expressions of MDR1, MRP1 genes in Tca8113 significantly descended at 4 h and 24 h post-hyperthermia （P<0.05）, but there was not statistical difference between 4 h and 24 h post-hyperthermia. GST-π expression had not statistical descent at 4 h, but there was significantly descend at 24 h post-hyperthermia（P<0.01）. Drug tolerance decreased and hyperthermia increased intracellular drug concentration in Tca8113/CBDEA and Tca8113（P<0.01）.Conclusion  Hyperthermia enhance chemotherapy effect and reverse MDR genes expression in tongue squamous cell carcinoma cell line. It is suggested that hyperthermia could be used as a method to overcome MDR, which is worth more investigation.