Background: HIV pre-exposure prophylaxis (PrEP) is a key component of HIV combination prevention strategies, which has the potential to decrease the incidence of HIV in men who have sex with men (MSM). This study aimed to evaluate the levels of PrEP acceptability and explore the factors associated with willingness to use PrEP among MSM in Liuzhou, China.Methods: Between November 2017 and May 2019, a cross-sectional survey was conducted among 829 MSM. The study participant were recruited through nonprobability sampling method. Participants’ demographics, HIV/AIDS knowledge, sexual behaviors, drug use and HIV test history were collected. Multivariate logistic regression was performed to identify factors associated with willingness to use PrEP.Results: A total of 829 MSM completed the survey, and 30.28%(95%CI: 27.3-33.4) were willing to use PrEP. In multivariate logistic regression, factors associated with a higher willingness to use PrEP include, being Zhuang or Han ethnic origin, recruited through peer introduction or gay venues, pursing higher education level, previous HIV testing and having oral sex with a man. However, had a casual sex in the past six months were associated with lower willingness to use PrEP.Conclusions: MSM in Liuzhou reported a relatively low level of willingness to use PrEP. The results indicate the need for effective education, targeted intervention, and define implementation strategies to promote PrEP acceptance among MSM.