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Posttranslational histone modification plays an important role in tumorigenesis. Histone modification is a dynamic response of chromatin to various signals, such as the exposure to calcitriol (1&#945;,25(OH)₂D₃). Recent studies suggested that histone modification levels could be used to predict patient outcomes in various cancers. Our study evaluated the expression level of histone 3 lysine 4 trimethylation (H3K4me3) in a cohort of 156 epithelial ovarian cancer (EOC) cases by immunohistochemical staining and analyzed its correlation to patient prognosis. The influence of 1&#945;,25(OH)₂D₃ on the proliferation of ovarian cancer cells was measured by BrdU proliferation assay in vitro. We could show that higher levels of H3K4me3 were correlated with improved overall survival (median overall survival (OS) not reached vs. 37.0 months, <i>p</i> = 0.047) and identified H3K4me3 as a potential prognostic factor for the present cohort. Ovarian cancer cell 1&#945;,25(OH)₂D₃ treatment induced H3K4me3 protein expression and exhibited antiproliferative effects. By this, the study suggests a possible impact of H3K4me3 expression on EOC progression as well as its relation to calcitriol (1&#945;,25(OH)₂D₃) treatment. These results may serve as an explanation on how 1&#945;,25(OH)₂D₃ mediates its known antiproliferative effects. In addition, they further underline the potential benefit of 1&#945;,25(OH)₂D₃ supplementation in context of ovarian cancer care.