<p>Hemodialysis treatment, along with its therapeutic effects, can cause many physical, psychological, social, and spiritual problems including stress, anxiety and depression in patients. The aim of this study is to determine the relationship of spiritual orientation with depression and hopelessness in patients receiving hemodialysis treatment. This descriptive and correlation study included 72 patients undergoing hemodialysis. Data were collected using a patient information form, the Spiritual Orientation Scale, the Beck Depression Inventory, and the Beck Hopelessness Scale. It was determined that the depression and hopelessness levels of the patients were moderate and their spiritual orientation was high. A statistically significant negative correlation was observed between spiritual orientation and hopelessness (<i>r</i> =  − 0.255, <i>p</i> = 0.030), while there was a statistically significant negative linear correlation between spiritual orientation and depression (<i>r</i> =  − 0.432, <i>p</i> = 0.001). The regression analysis revealed that spiritual orientation and other independent variables accounted for 20.6% of the variance in depression (<i>F</i>: 4.348, <i>p</i>: 0.003) and spiritual orientation accounted for 6.5% of the total variance in hopelessness (<i>F</i>(1, 70) = 4.885, <i>p</i> &lt; 0.05). It is recommended to evaluate patients undergoing hemodialysis in terms of spiritual orientation, hopelessness and depression, and to support their spiritual needs. Healthcare professionals should determine patients’ hopelessness and depression levels and predictive factors and make interventions to reduce them. Patients’ spiritual needs should be supported.</p>