Circular RNA mmu_circ_0001775 knockdown improves the osteogenic ability of mouse bone marrow mesenchymal stem cells
Dong Yi1, Shan Shuai1, Liu Jialin1, Han Xiangzhen1, He Huiyu1, 2
1Department of Prosthodontics and Implant, First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; 2Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
Abstract: BACKGROUND: In recent years, the use of circular RNA (circRNA) to regulate stem cell differentiation has become a new research focus. Therefore, loading circRNA on bone marrow mesenchymal stem cells to study their osteogenic properties is of great significance for the development of new materials to repair bone defect.
OBJECTIVE: To investigate the osteogenic effect of circRNA mmu_circ_0001775 knockdown on bone marrow mesenchymal stem cells.
METHODS: Bioinformatics analysis tools speculated that the upstream circRNA most closely related to miR-335-5p was mmu_circ_0001775. Lentivirus and negative control virus knockdown mmu_circ_0001775 were transfected into bone marrow mesenchymal stem cells. qRT-PCR was used to detect the expression levels of Runt-related transcription factor 2, osteocalcin, miR-335-5p, and mmu_circ_0001775 genes at 7 and 14 days of osteogenic induction, respectively, to verify their osteogenic effects. Western blot assay was used to detect the expression levels of bone morphogenetic protein 2 and osteopontin.
RESULTS AND CONCLUSION: The expression levels of Runt-related transcription factor 2 and osteocalcin at each time point in LV-mmu_circ_0001775-BMMSCs group were higher than those in LV-BMMSCs group (P < 0.01). The relative expression levels of bone morphogenetic protein 2 and osteopontin proteins were significantly higher in the LV-mmu_circ_0001775-BMMSCs group than those of LV-BMMSCs group (P < 0.05). The results showed that cirCRNA mmu_circ_0001775 knockdown could enhance the osteogenic ability of bone marrow mesenchymal stem cells.
Key words: circRNA, microRNA, osteogenic differentiation, jaw defect, tissue regeneration, bone marrow mesenchymal stem cells, molecular sponge