Background Overactive bladder (OAB) is a popular distressing health condition that has negative impact on health-related quality of life (HRQoL) of the inflicted individuals. This multicenter study was conducted to determine the prevalence of OAB symptoms and their impact on the HRQoL of medical and dentistry students. Methods This study was conducted in a cross-sectional design in the 3 main universities in Palestine. In addition to the sociodemographic, health, and academic characteristics of the medical and dentistry students, the questionnaire also contained the OAB symptom bother (6-items) and HRQoL (13-items) Short-Form (OAB-q SF) scales. Kruskal–Wallis test, Mann–Whitney U test, Pearson Chi-Square/Fisher's Exact Test, Spearman’s rank correlations, and a multiple linear regression model were used to analyze the data. Results Responses were collected from medical and dentistry students (n = 402). The median OAB symptom bother score was 54.1 [44.8, 81.9] and the median HRQoL score was 94.4 [88.4, 94.4]. There was a strong negative correlation between the OAB and HRQoL scores (Spearman’s rho = 64.4%, p value < 0.001). OAB scores were significantly higher among dentistry students, females, who had chronic disease, and those who reported stressful life. HRQoL scores were significantly higher among medicine students, those who reported less stressful life, and those who reported satisfaction with their social life. Dentistry students, female, and those who self-reported high stress were 1.94-fold (95% CI 1.05, 3.56), 1.91-fold (95% CI 1.16, 3.14), and 1.88-fold (95% CI 1.21, 2.91) more likely to report less than optimal HRQoL compared to medicine students, male, and those who self-reported low stress, respectively. Conclusions Our findings suggested that OAB symptoms were prevalent among medical and dentistry students across Palestinian universities. Decision makers in academia, healthcare authorities, and advocacy groups might need to design appropriate interventions to address health and wellbeing issues of medical and dentistry students. Using appropriate diagnostic procedures, reducing stress, and improving the social life might help in reducing the burden on OAB and improve the HRQoL of medical and dentistry students. More investigations should be conducted to investigate if such interventions are effective in reducing OAB symptoms and improving HRQoL.