Display omitted •Shift work disrupts circadian rhythms, and night-shift work is associated with the occurrence of substance use disorders; however, there is no evidence to support a link between night-shift work and behavioral addictions.•In this cross-sectional study using an Internet survey, we examined the association between shift work and gambling, a potential risk behavior for behavioral addictions, among workers (n = 21,134), and the results showed an association between night-shift work and gambling participation (adjusted odds ratio aOR, 1.39, 95 % confidence interval CI, 1.25–1.53, p < 0.001).•When restricted to respondents with past-year gambling experience (n = 9,739), night-shift work was associated with problem gambling, defined as the problem gambling severity index score of 8 or greater (aOR 1.94, 95 % CI, 1.94–2.40, p < 0.001).•Night-shift work was associated with gambling in workers and with problem gambling among working and gambling respondents, suggesting that gambling might be a factor associated with shift work-related harms. Night-shift work disturbs sleep and is associated with poor health conditions among workers. We aimed to investigate the association between night-shift work and gambling among workers and the association between night-shift work and problem gambling in working and gambling participants. This cross-sectional study used data from an online survey conducted between February 6 and 27, 2023 in Japan. A total of 21,134 workers participated in this study, including 9,739 respondents who had gambled in the past year. We estimated the association between night-shift work and gambling among workers and the association between night-shift work and problem gambling among those who gambled at the survey. We defined problem gambling as a score ≥ 8 on the Problem Gambling Severity Index. All estimates were weighted using a nationally representative survey in Japan. We fitted multivariable weighted logistic regression models after adjusting for 14 confounders. The weighted prevalence of gambling among non-night and night-shift workers was 42.1 % and 55.4 %, respectively. When focusing on workers gambling in the survey, the prevalence of problem gambling among non-night and night-shift workers was 8.8 % and 24.2 %, respectively. The weighted multivariable logistic regression analyses showed that night-shift work was associated with gambling participation among workers (adjusted odds ratio aOR, 1.39, 95 % confidence interval CI 1.25–1.53, p < 0.001). In addition, night-shift work was associated with problem gambling among those who gambled (aOR 1.94, 95 % CI 1.57–2.40, p < 0.001). Night-shift work was associated with gambling among workers and with problem gambling among those who gambled.