Stress is strongly associated with depression development and leaves traces in the epigenetic regulation of gene transcription. The oxytocin system has been implicated in allostatic processes promoting adaption to environmental stressors. Therefore, interactions of the oxytocin system with the environment, e.g., in form of methylation status of the gene coding for oxytocin, is a logical candidate for the investigation of the biological underpinnings of depression. Recently, we found hypomethylation of the oxytocin gene in patients suffering from depression as compared to healthy controls rendering a role for the oxytocin system in depression course likely. However, since the replicability of findings is a key point of criticism in (epi-)genetic research, we aimed at replicating our previous findings in a preregistered study in a new sample of N = 85 patients with MDD diagnosis and N = 85 healthy controls. We investigated DNA-methylation of the oxytocin gene, stressful life events and depression severity. As expected and in accordance with our earlier study, we found significant hypomethylation of the oxytocin gene – patients had lower methylation levels than controls – with regard to mean methylation and on the level of single CpG units. Methylation was not associated with critical life events, even though patients reported significantly more and more traumatic stressful life events in comparison to healthy controls. This study shows that hypomethylation of the oxytocin gene can be demonstrated in a reproducible fashion and provides further evidence for the involvement of the oxytocin system in depression.