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This study developed a radiation protection education program and verified the effects of the program on the knowledge, attitude, and behavior of operating room nurses regarding radiation protection. A quasiexperimental design with non-equivalent control group pretest-posttest was used. The conceptual framework was Keller’s ARCS (Attention, Relevance, Confidence, and Satisfaction) model. There were 21 participants in the experimental group and 21 in the control group. The experimental group attended four sessions of the radiation protection education program while the control group was given printouts as learning materials. The learning program lasted for a total of 80 minutes over one week and consisted of four sessions of 20 minutes each. The data were analyzed using repeated measures ANOVA, independent t-test, Friedman test, Wilcoxon signed-rank test and Mann-Whitney U test with SPSS WIN 23.0. Results: There was a significant difference between the experimental group and the control group in the attitude (p= .001) and behavior (p<.001) of radiation protection. Also the experimental group showed significantly lower posttest scores in the value of the personal radiation exposure dosimeter using TLD (thermoluminescence dosimeter) than control group (p=.028) The web-based radiation protection program demonstrated statistically significant improvements in nurses’ attitudes and behaviors compared to the control group. However, its effect on knowledge acquisition was not clearly distinguishable, as both the web-based intervention and leaflet yielded comparable outcomes in knowledge improvement. Owing to its high accessibility, flexibility, and capacity for self-directed and repetitive learning, the web-based program presents itself as a feasible and effective educational intervention for nurses requiring continuous professional development. Further research is needed to enhance knowledge acquisition.