In photodynamic therapy (PDT), oxygen plays important role. Because of singlet oxygen which is produced by activated photosensitizer after laser irradiation of specific wavelength. The aim of this study is to find how oxygen concentration affects anticancer effect in PDT. Groups were divided into PDT with oxygen applied group and only PDT applied group. PDT with oxygen applied group supplied oxygen for 15 minute before laser irradiation. In vitro, CT-26 cell was incubated with various concentration of photofrin (50.0~0.05 ㎍/ml) and was irradiated with 632 ㎚ diode laser 6 hr after application of photofrin. The cell viability of two groups was assessed by MTT assay. In vivo, CT-26 cell line was transplanted into the subcutaneous tissue of BALB/c mouse. The anticancer effect of two groups was measured by tumor volume change. In vitro study, the cell viability was significantly decreased at 1.56~3.13 ㎍/ml in PDT with oxygen applied group. In vivo study, the PDT with oxygen applied group significantly higher reduction rate of tumor volume 7 days after PDT compared to PDT only group. The high oxygen concentration might enhance the anticancer effect of the photodynamic therapy.