[Objectives]  To determine the role of ace-miR-3759-y in target gene expression and body weight of Apis cerana cerana larvae in order to provide a foundation for further study of the ace-miR-3759-y-regulated molecular mechanism underlying larval development. [Methods]  The expression and sequence of ace-miR- 3759-y were confirmed by Stem-loop RT-PCR and Sanger sequencing. A. cerana cerana larvae were fed Mimic and Inhibitor to overexpress and knockdown ace-miR-3759-y in larval hemolymph. RT-qPCR was then used to determine the effect of ace-miR-3759-y overexpression and knockdown on the target genes CIC and CKI, and on larval weights which were measured using electronic scales. [Results]  ace-miR-3759-y was both present and expressed in the hemolymph of A. cerana cerana larvae. Treatment with Mimic significantly up-regulated its expression in 4- and 5-day-old larvae relative to the control group. It was also upregulated in 6-day-old larvae, but this was not significant. Conversely, treatment with Inhibitor significantly down-regulated ace-miR-3759-y expression in 4-, 5- and 6-day-old larvae relative to the control. After overexpression of ace-miR-3759-y, the expression of CIC in 4-, 5- and 6-day-old larvae was significantly down-regulated, whereas that of CKI was significantly down-regulated in 4- and 5-day-old larvae. It was also down-regulated in 6-day-old larvae, but not significantly so. Knockdown of ace-miR-3759-y significantly up-regulated the expression of CIC in 5-day-old larvae. Expression was also up-regulated in 4- and 6-day-old larvae but this was not significant. Knockdown of ace-miR-3759-y also up-regulated CKI in 4-, 5- and 6-day-old day-old larvae, although the degree of upregulation in the latter two age classes was not significant. Moreover, treatment with Mimic caused a non-significant decrease in the body weight of 6-day-old larvae relative to the Mimic control group and treatment with Inhibitor caused a non-significant increase in the body weight of 6-day-old larvae relative to the Inhibitor control group. [Conclusion]  Effective overexpression and knockdown of ace-miR-3759-y in A. cerana cerana larvae can be achieved by feeding mimic and inhibitor. ace-miR-3759-y negatively regulates expression of CIC and CKI but has insignificant effects on body weight.