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2023, Vol. 27 ›› Issue (19): 3023-3028

miR-889-3p inhibits osteogenic differentiation of adipose-derived mesenchymal stem cells by targeting Runx2

Zhang Yaotian1, Cui Jun2, Liu Jingyi2   

  1. 1Shenyang Medical College, Shenyang 110000, Liaoning Province, China; 2Affiliated Central Hospital of Shenyang Medical College, Shenyang 110000, Liaoning Province, China

  • Received:2022-06-08 Accepted:2022-07-28 Online:2023-07-08 Published:2022-11-28

  • Contact: Cui Jun, Master, Affiliated Central Hospital of Shenyang Medical College, Shenyang 110000, Liaoning Province, China

  • About author:Zhang Yaotian, Shenyang Medical College, Shenyang 110000, Liaoning Province, China

  • Supported by:

    Guidance Program of Liaoning Provincial Natural Science Foundation, No. 20180550071 (to CJ)


Abstract: BACKGROUND: Studies have shown that miR-889-3p is able to participate in the regulation of the proliferation and differentiation of a variety of cells, but its effect on the osteogenic differentiation of adipose-derived mesenchymal stem cells remains unclear.  
OBJECTIVE: To investigate the regulation of miR-889-3p in osteogenic differentiation of adipose-derived mesenchymal stem cells.
METHODS: Adipose-derived mesenchymal stem cells from SD rats were isolated and cultured in vitro, transfected with miR-889-3p mimics, Mir-889-3p inhibitor and negative control (miR-NC) then induced osteogenic differentiation. RT-PCR and western blot assay were used to detect alkaline phosphatase activity, Runx2, osteocalcin, osteopontin, Osterix and other osteogenic related mRNA and protein expression levels. Adipose-derived mesenchymal stem cells were co-transfected with wild-type Runx2 and mutant Runx2 with miR-889-3p mimic and miR-NC, respectively. Luciferase reporter gene assay was used to detect the targeting relationship between miR-889-3p and Runx2.  
RESULTS AND CONCLUSION: (1) The expression level of miR-889-3p decreased gradually with the prolongation of osteogenic induction. (2) The number, size, and color depth of mineralized nodules in the miR-889-3p inhibitor group were significantly higher than those in the miR-889-3p mimic group and miR-NC group on days 7 and 14 of osteogenic induction (P < 0.05). (3) Overexpression of miR-889-3p significantly suppressed alkaline phosphatase activity, Runx2, osteocalcin, osteopontin, Osterix mRNA and protein expression, whereas the knockdown of miR-889-3p remarkably enhanced osteogenesis-related mRNA and protein expression. (4) The results of dual-luciferase reporter gene assay showed that overexpression of miR-889-3p significantly reduced the luciferase activity of wild-type Runx2. (5) These results suggest that miR-889-3p negatively regulates osteogenic differentiation of adipose-derived mesenchymal stem cells by targeting Runx2.

Key words: miR-889-3p, Runx2, adipose-derived mesenchymal stem cell, osteogenic differentiation, bone defect


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Chinese Association of Rehabilitation Medicine

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