Objective To develop a more convenient and stable method for assessment of drug sensitivity of prostate cancer based on real-time cell analysis system as a reference for clinical treatment. Methods Human prostate cancer VCaP, DU145, PC-3, PC-3M-2B4 and PC-3M-IE8 cells were chosen to detect the sensitivity to three drugs, docetaxel, cabazitaxel and abiraterone acetate. Serial dilutions of the three drugs were used to treat the cell culture for 24 hours. The drug-induced effects on the cell lines after an incubation of 24 hours were recorded by the real-time cell analysis system to determine the half maximal inhibitory concentration (IC50). Results Docetaxel showd IC50 of 8.81 nmol/L, 11.61 nmol/L, 1.78 nmol/L, 1.44 nmol/L, 8.69 nmol/L for VCaP, DU145, PC-3, PC-3M-2B4, PC-3M-IE8 cells, respectively. Cabazitaxel showed IC50 of 3.73 nmol/L, 3.96 nmol/L, 10.41 nmol/L, 5.43 nmol/L, and 7.37 nmol/L, respectively, for the five cell lines. Abiraterone acetate showed IC50 of 8.34 μmol/L, 8.60 μmol/L, 24.20 μmol/L, 8.59 μmol/L, and 13.21 μmol/L for the five cell lines. Conclusions PC-3M-2B4 and DU145, VCaP and PC-3 cells can be used as control for docetaxel, cabazitaxel and abiraterone acetate to establish cell models for the drug screening in vitro and to provide reference for clinical applications.