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Background: Lack of physical activity and unhealthy eating behavior are considered to be global issues and lead the risk of non-communicable diseases. The previous studies indicated that mobile applications have the potential for delivering health information to promote health behaviors. Objective: To evaluate the effect of health promotion intervention through smartphone LINE application to increase knowledge, perception, and practice of physical activity and healthy eating behavior among overweight women in urban community. Materials and Methods: A quasi-experimental study was conducted among 100 overweight women with a BMI between 25 and 29.9 kg/m², and aged between 35 and 65 years living in urban community in Bangkok between October 2017 and April 2018. Participants were selected to the intervention group and control group. The intervention program consisted of group education, self-monitoring, individual counseling, and smartphone LINE application group chatting about physical activity and healthy eating behavior. Data were collected at baseline and after the intervention ended at 24 weeks. Chi-square, paired samples t-test, and independent t-test were used to find the effect of the program. Results: At the end of 6-month, the results indicated that the intervention group had significantly increased in mean of knowledge (p<0.001) and perception (p<0.001) of physical activity and healthy eating behavior, food frequency scores (p<0.001), MET-physical activity (p=0.007) and decreased in mean of time spent sitting (p<0.001), body weight (p=0.043), and BMI (p<0.001) when compared to the control group. Conclusion: The use of LINE application was found effective to deliver health information, share and exchange their experienced, motivate, and monitor their physical activity and healthy eating behavior. This app can help the participants to increase the physical activity and healthy eating behavior, and to decrease their sedentary behavior resulting in weight loss. It is useful for healthcare providers to promote the health behaviors. [ABSTRACT FROM AUTHOR]