The safety climate is becoming more and more important in the processes of subway operation safety management due to various accidents. The research objective of this study is to explore the effects of safety climate and personal factors on safety behavior in subway operation. First, a conceptual model is developed based on the literature review and expert experience. Then, data are collected from 352 workers in the Xuzhou subway operation company by questionnaire survey. Third, the structural equation model is employed to do model analysis based on SPSS and AMOS, and the final model is achieved through a serious of model tests and modification. At last, the quantitative effect of safety climate on worker’s safety behavior in subway operation is obtained and analyzed. The results show that the descending order of total influence effect of safety climate on safety behavior is safety attitude (0.36), safety communication (0.265), safety policy (0.238), safety education and training (0.1), management commitment (0.099), and safety participation (0.073), respectively. The total influence effects of mediator variables (safety awareness and safety ability) are 0.242 and 0.194, respectively. This study would be beneficial by offering recommendations in regard to worker’s safety behavior to raise the safety level in subway operation.