is an animal model for studying the reproduction and development of crustaceans. In this study, we knocked down the gene by injecting dsRNA into and inhibited gene expression by injecting PLK1 inhibitor BI6727 into . Then, the cell proliferation level, apoptosis level, and PI3K/AKT signaling expression level were detected. Our results showed that the proliferation level of spermatogenic cells decreased, while the apoptosis level increased after knockdown or inhibition. In order to verify whether these changes are caused by regulating the PI3K/AKT pathway, we detected the expression of PI3K and AKT proteins after knockdown or inhibition. Western Blot showed that in both the knockdown group and the inhibition group, the expression of PI3K and AKT proteins decreased. In addition, immunofluorescence showed that Es-KIF2A and Es-PLK1 proteins were co-localized during spermatogenesis. To further explore the upstream and downstream relationship between Es-KIF2A and Es-PLK1, we detected the expression level of Es-PLK1 after knockdown as well as the expression level of Es-KIF2A after inhibition. Western Blot showed that the expression of Es-PLK1 decreased after knockdown, while there was no significant change of Es-KIF2A after inhibition, indicating that Es-PLK1 may be a downstream factor of Es-KIF2A. Taken together, these results suggest that Es-KIF2A upregulates the PI3K/AKT signaling pathway through Es-PLK1 during the spermatogenesis of , thereby affecting the proliferation and apoptosis levels of spermatogenic cells.