Estimation of the relative biological effectiveness (RBE) of the proton beam at the National Cancer Center Proton Therapy Center in Korea (NCCPTC) is required clinically for the treatment of cancer. The proton beam was fixed at 190 MeV with 6 cm a spread out Bragg peaks (SOBP) for which corresponds to most frequent clinical condition. The RBE was estimated from the survival of human salivary gland (HSG) cells using the traditional colonogenic and MTT assays. The HSG cells were also irradiated in a cell-stack chamber and monitored for survival to identify whether the characteristic depth-dependent survival pattern was observed. The RBE of the NCCPTC was estimated to be 1.024 +/- 0.007 and 1.049 +/- 0.028 at the middle of SOBP using colonogenic and MTT assays, respectively. Further analysis of the biological response of proton exposure revealed no difference compared to conventional X-ray treatment in western blot, and FACS analysis. The proton beam of the NCCPTC also exhibited the characteristic depth-dependent survival pattern. The estimated RBE value of NCCPTC was slightly smaller than generic RBE value of 1.1 for protons of the majority of centers. Due to the recommendation of a generic RBE of 1.1 for protons, a representative RBE value of 1.1 was assigned for clinical application for proton beams at the NCCPTC.