Background: Adolescent substance use has been widely related to different individual, school, family and commu-nity factors. Yet, the number of studies with all these variables together in a model from an ecological perspective is still low, and they rarely used a longitudinal design. The aim of this study was to explore, from an ecological perspective, the prospective impact of different individual, school, family and neighbourhood factors on adoles-cent substance use.Methods: This was a longitudinal study with a one-year follow up. There were 881 participants (Mage = 12.57; 48.1% females) at wave 1, of which 686 (Mage = 13.51; 51.8% females) were followed-up at wave 2. Validated questionnaires were used for data collection.Results: Regression analyses showed that higher substance use was predicted by high family socio-economic status cross-sectionally and longitudinally, and it was related to low neighbourhood socio-economic status cross -sectionally only. Participants who disliked school and had a poor academic performance were more likely to use substances, both cross-sectionally and longitudinally.Conclusions: The inclusion of families in substance use prevention programmes could be a key component in these interventions. Moreover, promotion of a positive school climate could protect adolescents from using substances.