It is recognised that cultural factors play a role in the onset and continuation of several mental health problems. However, there is a significant lack of empirical studies investigating the relationships between cultural factors and gambling behavior. This study assessed whether the subjective cultures through which subjects interpret and enact their experience of the social environment play a major role in increasing (or decreasing) the probability of pathological gambling. Participants, recruited in three different contexts (public health services for the treatment of addiction, casino, undergraduate course) were subjected to the South Oaks Gambling Screen (SOGS) (Lesieur and Blume in Am J Psychiatry 144(9):1184-1188, 1987), in order to identify a group of pathological gamblers-and with the Questionnaire on the Interpretation of the Social Environment (QUISE) (Mossi and Salvatore in Eur J Educ Psychol 4(2):153-169, 2011)-in order to detect their subjective cultures. The study compares pathological group (scoring >5 on SOGS, n = 34) and a healthy control group (scoring <1 on SOGS, n = 35). One-way analysis of variance (ANOVA) was used to compare groups on QUISE scores of subjective culture. Moreover, a logistic regression was applied in order to esteem the capability of the QUISE scores to differentiate between pathological gamblers and control. The results are consistent with the hypothesis that pathological group expresses different subjective cultures compared with no gambler subjects. The theoretical and clinical implications of the results are discussed.