Cervical cancer is the second most common female malignancy worldwide. Although Cisplatin (DDP) is a commonly used chemotherapy agent for cervical cancer, the development of drug resistance has limited its clinical use. Fluoxetine (FLT), which is widely used to treat depression, has exhibited potent anticancer activity against different cancer cell types. Until now, the effect of Fluoxetine on cervical cancer cells has not been previously studied. The aim of this study was to evaluate the anticancer effect of FLX alone or combing with DDP on human cervical cancer cell, as well as its molecular mechanism. Here, we found that FLX combining with DDP could significantly inhibit the proliferation of HeLa cells in a dose dependent manner, and induce cell cycle arrest at G0/G1 phase and apoptosis in vitro. Moreover, FLX and DDP in combination could suppress the tumor growth in a HeLa xenograft mouse model in comparison to FLX or DDP alone. Molecular mechanism demonstrated FLX combining with DDP not only could suppress the levels of multi-drug resistance genes including GST-p and P-gp, but also affect the levels of apoptotic related genes including down-regulation of the Bcl-2 and up-regulation of caspase-9 and p17. In conclusion, FLX is a highly effective chemosensitizer, and FLX in combination with DDP might be an effective treatment against cervical cancer.