IntroductionNegative affect was identified as an important barrier to smoking cessation. Three-part breathing exercise showed a significant effect on decreasing negative affect immediately after being practiced. Thus, this study evaluated the effect of three-part breathing exercise on smoking cessation. MethodsA 6-month cluster-randomized clinical trial was conducted. Forty-three participants recruited from 8 companies in Bangkok Metropolitan areas were randomly assigned at the cluster level into either the intervention or control groups. Control group (n = 23) received counseling for smoking cessation once a week for 12 weeks. Intervention group (n = 20) received counseling for smoking cessation plus a three-part breathing exercise program once a week for 12 weeks. The primary outcomes were 7-day point prevalence and continuous abstinence rate as validated by saliva cotinine. The secondary outcomes were cigarette cravings, nicotine withdrawal symptoms, affect and quality of life. ResultsThe results revealed no significant difference in smoking abstinence rate between the three-part breathing exercise and control group. Participants demonstrated significant pre-post improvement in cigarette cravings, nicotine withdrawal symptoms, affect, and quality of life within each group. ConclusionThere were no statistically significant differences between the two groups. However, the improvement in abstinence rate from the three-part breathing exercise was deemed clinically relevant. Thus, it may be recommended to smokers interested in smoking cessation and more research is needed on this topic.