Abstract:Objective: To investigate the effects of LncRNA H19 targeting miR-19b-3p/SOX9 pathway on the osteogenic differentiation of adipose-derived mesenchymal stem cells (ADMSCs). Methods: Human ADMSCs were cultured and identified in vitro. The cells were divided into five groups: control group (regular culture), H19 group (transfected with LncRNA H19 lentivirus overexpression vector), LncRNA-NC group (transfected with lentivirus empty vector), miR-19b-3p group (co-transfected with LncRNA H19 lentivirus overexpression vector and miR-19b-3p mimic lentivirus vector), and miR-NC group (co-transfected with LncRNA H19 lentivirus overexpression vector and miR-NC lentivirus vector). The expression of LncRNA H19, miR-19b-3p, SOX9, zinc finger osteoblast-specific transcription factor (Osx), osteocalcin (OCN), and RUNX2 was detected in each group. ALP staining and Alizarin Red staining were observed in each group, cell proliferation activity and ALP activity were compared, and the expression of SOX9, BMP-2, and OPN proteins in each group was analyzed to verify the targeted relationship between LncRNA H19 and miR-19b-3p. Results: Compared with the control group and LncRNA-NC group, the expression of LncRNA H19 and SOX9, Osx, OCN, and RUNX2 mRNA, as well as the expression of SOX9, BMP-2, and OPN proteins, ALP activity, and the amount of mineralized nodule formation were increased in the H19 group, while the expression of miR-19b-3p was decreased (P<0.05). Compared with the H19 group and miR-NC group, the expression of LncRNA H19 and SOX9, Osx, OCN, and RUNX2 mRNA, as well as the expression of SOX9, BMP-2, and OPN proteins, ALP activity, and the amount of mineralized nodule formation were decreased in the miR-19b-3p group, while the expression of miR-19b-3p was increased (P<0.05). There was no statistical difference in the absorbance of each group at 24h, 48h, and 72h (P>0.05). There was a targeted relationship between LncRNA H19 and miR-19b-3p. Conclusion: LncRNA H19 can negatively regulate miR-19b-3p, upregulate SOX9, and contribute to the differentiation of human ADMSC cells into osteoblasts. LncRNA H19 can negatively regulate miR-19b-3p, upregulate SOX9, and promote the osteogenic differentiation of human ADMSCs.
[1] Fu X,Liu G,Halim A,et al.Mesenchymal stem cell migration and tissue repair[J].Cells,2019,8(8):1-18. [2] An Y,Lin S,Tan X,et al.Exosomes from adipose-derived stem cells and application to skin wound healing[J].Cell Prolif,2021,54(3):1-12. [3] Yu H,Huang Y,Yang L.Research progress in the use of mesenchymal stem cells and their derived exosomes in the treatment of osteoarthritis[J].Ageing Res Rev,2022,80(1):1-4. [4] Zhou Z,Hossain MS,Liu D.Involvement of the long noncoding RNA H19 in osteogenic differentiation and bone regeneration[J].Stem Cell Res Ther,2021,12(1):1-9. [5] Qian L,Pi L,Fang BR,et al.Adipose mesenchymal stem cell-derived exosomes accelerate skin wound healing via the LncRNA H19/miR-19b/SOX9 axis[J].Lab Invest,2021,101(9):1254-1266. [6] Liu D,Wang B,Qiu M,et al.MiR-19b-3p accelerates bone loss after spinal cord injury by suppressing osteogenesis via regulating PTEN/Akt/mTOR signalling[J].Cell Mol Med,2021,25(2):990-1000. [7] Meng X,Zhu H.SOX9 inhibits the progression of osteonecrosis of the femoral head via the activation of the wnt/beta-catenin pathway[J].Invest Surg,2023,36(1):1-8. [8] Sumarwoto T,Suroto H,Mahyudin F,et al.Role of adipose mesenchymal stem cells and secretome in peripheral nerve regeneration[J].Ann Med Surg (Lond),2021,67(1):1-10. [9] Cheng M,Yuan W,Moshaverinia A,et al.Rejuvenation of mesenchymal stem cells to ameliorate skeletal aging[J].Cells,2023,12(7):1-21. [10] Ge J,Yu YJ,Li JY,et al.Activating Wnt/β-catenin signaling by autophagic degradation of APC contributes to the osteoblast differentiation effect of soy isoflavone on osteoporotic mesenchymal stem cells[J].Acta Pharmacol Sin,2023,44(9):1841-1855. [11] Bi HU,Wang D,Liu X,et al.Long non-coding RNA H19 promotes osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by regulating microRNA-140-5p/SATB2 axis[J].Biosci,2020,45(1):1-13. [12] He Q,Li R,Hu B,et al.Stromal cell-derived factor-1 promotes osteoblastic differentiation of human bone marrow mesenchymal stem cells via the LncRNA-H19/miR-214-5p/BMP2 axis[J].Gene Med,2021,23(9):1-25. [13] Xiaoling G,Shuaibin L,Kailu L.MicroRNA-19b-3p promotes cell proliferation and osteogenic differentiation of BMSC by interacting with LncRNA H19[J].BMC Med Genet,2020,21(1):1-8. [14] Della Bella E,Buetti-Dinh A,Licandro G,et al.Dexamethasone induces changes in osteogenic differentiation of human mesenchymal stromal cells via SOX9 and PPARG,but not RUNX2[J].Int Mol Sci,2021,22(9):1-16.