This study aims to investigate the potential association between use of WhatsApp in healthcare settings for work-related purposes and depression, stress, and anxiety levels among a sample of healthcare workers in Saudi Arabia. Methods: This was a cross-sectional study of healthcare employees working in different hospitals in Jazan. The data was collected via a semi-structured, self-administered questionnaire with three components that measured the demographics of the sample; the presence of depression, anxiety, and stress; and their use of WhatsApp in the work environment. A multivariate regression analysis was carried out to estimate the statistical likelihood of depression, anxiety, and stress as a result of using WhatsApp and how these factors affected occupational and social relationships. Results: A total of 294 healthcare workers participated in the current study. The median age of the participants was 32 years, and the gender distribution was almost even. Over 90% of the participants stated that they were members of work-related WhatsApp groups, and nearly 70% agreed that use of WhatsApp in work-related settings could be stressful. Among the recruited sample, 48.6% exhibited abnormal levels of depression, 55.8% had abnormal levels of anxiety, and 63% showed abnormal levels of stress. The findings of the regression analysis (P values <0.05) indicated that the likelihood of these participants suffering with depression, anxiety, and stress was high, with the participants themselves agreeing that using WhatsApp in work settings could be stressful and affect their relationships with colleagues, friends, and family members. Conclusion:The findings suggests the presence of a potential association between use of WhatsApp for work-related purposes and levels of depression, anxiety, and stress, especially among those who perceive its use as a stressor and influencer on occupational and social relationships.