[Aim]  The objective of this study is to investigate the effect of ace-miR-14-y overexpression on expression of target genes in the Apis cerana cerana worker larvae as well as weights of individual and gut. [Methods]  Stem-loop RT-PCR and Sanger sequencing were utilized to confirm the expression and sequence of ace-miR-14-y. Relative software were used to predict and analyze the target genes of ace-miR-14-y. Overexpression of ace-miR-14-y in the larval gut was performed by feeding mimic, followed by examination of the effect of overexpression and the relative expression level of target genes after overexpression. Electronic balance was employed to weigh the larval individual and gut. [Results]  The results demonstrated that ace-miR-14-y authentically existed and expressed in the A. cerana cerana worker larval gut. ace-miR-14-y targeted a total of 265 genes, involving 33 GO terms and 180 KEGG pathways. The expression level of ace-miR-14-y in the mimic-miR-14 group was significantly higher than that in the mimic-NC group (P<0.05). After overexpression of ace-miR-14-y, the expression level of target genes Wg and AP-1 were both significantly downregulated (P<0.05) and the weights of larval individual and gut were increased with extreme significance (P<0.01). [Conclusion]  The overexpression of ace-miR-14-y significantly affects the target genes Wg and AP-1’s expression in Apis cerana cerana worker larval gut as well as the weights of larval individual and gut, ace-miR-14-y plays a potential regulatory part through the regulation of expression of Wg and AP-1.