In order to explore the potential role of highly-expressed lncRNAs (HlncRNAs) in regulating the midgut development of Apis mellifera ligustica worker, the omics data obtained in the previous at 7- and 10-day-old A. m. ligustica workers’ midguts were used to screen out the HlncRNAs. RT-PCR was performed to confirm the real expression of aforementioned HlncRNAs and analysis the expression pattern of them during the developmental process of 1~21 days old workers’ midguts, and bioinformatic analyses of the regulatory function of HlncRNAs were further performed. The results suggested that the 9 HlncRNAs in the 7- and 10-day-old worker’ s midgut were the same. These HlncRNAs were expressed in 1- to 21-day-old workers’ midguts but their expression patterns were different. HlncRNAs regulated 32 upstream and downstream genes and 18 co-expressed mRNAs via cis-acting and trans-acting manner, respectively. Additionally, these HlncRNAs can target 55 miRNAs and further bind to 131 mRNAs. These results reveal the 9 HlncRNAs potentially regulated functional terms and pathways such as the stress response, sugar digestion and absorption through cis-acting, trans-acting and ceRNA network to regulate the development of A. m. ligustica worker’ s midgut.