Plant reproduction is a very important process on Earth from the perspective of biodiversity, biomass gain and crop productivity. It is therefore important to understand sex determination process and many researchers are investigating the molecular basis of this phenomenon. However, information on the influence of transcription factors (TFs) on this process is limited, although cucumber is a model plant in this regard. In the present study, based on RNA-seq data for differential gene expression (DEG) analyses, we aimed to investigate the regulatory TFs that may influence the metabolomic processes in the shoot apex containing the forming flower buds. Therefore, a robust TF database was established for the B10v3 cucumber genome. Sex-specific interactome network maps were generated, indicating the regulatory TFs by their effects on DEGs and further on processes leading to the formation of different sex flowers. The network analysis identified major families of regulatory TFs. The most abundant families were: MYB, AP2/ERF, NAC and bZIP, and those with the greatest impact on developmental processes were identified, namely the AP/ERF family, followed by DOF, MYB, MADS and others. Thus, the central nodes and key regulators in the networks were identified with respect to male, female and hermaphrodite. Here, we proposed the first model of the regulatory network of TFs that influences the metabolism of sex development in cucumber. These findings may help to understand the molecular genetics and functional mechanisms underlying sex determination processes.