Abstract:Objective To investigate the effect of miR-214-5p on osteoclastic differentiation and anti osteoporosis through PTEN expression. Methods Twenty patients with osteoporosis who were admitted in Tangdu Hospital of Air Force Military Medical University from March 2019 to May 2020 were chosen as osteoporosis group (n=20). miR-214-5p mimics (miR-214-5p mimics group), miR-214-5p empty plasmid (miR-214-5p vector), shZFAS1 vector (sh PTEN group) and shZFAS1 empty vector (SHNC group) were transfected into the cells. The control group (n=20)cells were not treated with any transfection. The mRNA levels of miR-214-5p and PTEN in bone marrow mononuclear cells and NFATc1, MMP9, TRAP and ctsk mRNA in RAW264.7 monocyte macrophages were detected by RT qPCR; the positive rate of TRAP staining was detected by TRAP staining; the protein levels of PTEN, NFATc1, Akt and PI3K were detected by Western blot. Results The expression of miR-214-5p in osteoporosis was significantly higher than that in control group (P< 0.001), and the expression of PTEN in osteoporosis was significantly lower than that in control group (P=0.009); the percentage of TRAP positive cells in miR-214-5p mimics group was significantly higher than that in control group and miR-214-5p vector group (P< 0.001); the percentage of TRAP positive cells in SH PTEN group was significantly higher than that in control group and sh PTEN group The levels of NFATc1, MMP9, TRAP and ctsk in miR-214-5p mimics group were significantly higher than those in control group (P<0.001); the levels of NFATc1, MMP9, TRAP and ctsk in SH PTEN group were significantly higher than those in control group (P<0.001); the percentage of TRAP positive cells in miR-214-5pmimics+PTEN group was significantly lower than that in miR-214-5p mimics group (P< 0.001). Conclusions The expression of miR-214-5p in bone marrow mononuclear cells of patients with osteoporosis is up-regulated, miR-214-5p can regulate the expression of PTEN, and overexpression of miR-214-5p may promote osteoclast differentiation by targeting PTEN.
Cummings S R,San Martin J,Mcclung M R,et al. Denosumab for prevention of fractures in postmenopausal women with osteoporosis.[J]. N Engl J Med, 2009, 361(8):756-765.
[2]
Christiansen C . Consensus development conference: diagnosis, prophylaxis, and treatment of osteoporosis[J]. Am J Med, 1993, 94(6):646-650.
[3]
Neer R M,Arnaud C D,Zanchetta J R, et al. Effect of parathyroid hormone (1-34) on fractures and bone mineral density in postmenopausal women with osteoporosis[J]. N Engl J Med, 2001, 344(19):1434-1441.
[4]
Materozzi M,Merlotti D,Gennari L,et al.The potential role of miRNAs as new biomarkers for osteoporosis[J]. Int J Endocrinol, 2018, 18(1):1-10.
[5]
Zhang M, Wang D, Zhu T,et al.miR-214-5p targets ROCK1 and suppresses proliferation and invasion of human osteosarcoma cells[J]. Oncol Res, 2017, 25(1):75-81.
[6]
Hsieh S C,Chen N T,Lo S H.Conditional loss of PTEN leads to skeletal abnormalities and lipoma formation[J]. Mol Carcinog, 2010, 48(6):545-552.
[7]
Yang G,Sun Q,Teng Y,et al. PTEN deficiency causes dyschondroplasia in mice by enhanced hypoxia-inducible factor 1α signaling and endoplasmic reticulum stress[J]. Development, 2008, 135(21):3587-3597.
Kaur M,Nagpal M,Singh M.Osteoblast-n-osteoclast: making headway to osteoporosis treatment[J]. Curr Drug Targets, 2020, 21(13):198-211.
[11]
Quinn J M W,Neale S,Fujikawa Y,et al. Human osteoclast formation from blood monocytes, peritoneal macrophages, and bone marrow cells[J]. Calcif Tissue Int, 1998, 62(6):527-531.
[12]
Shen G,Ren H,Qiu T,et al.Implications of the interaction between miRNAs and autophagy in osteoporosis[J]. Calcif Tissue Int, 2016, 99(1):1-12.
[13]
Krzeszinski J Y,Wei W,Huynh H D,et al.Authorcorrection: miR-34a blocks osteoporosis and bone metastasis by inhibiting osteoclastogenesis and Tgif2[J]. Nature, 2019, 570(7761):51-63.
[14]
Zhang Q,Zhang S. miR-214 promotes radioresistance in human ovarian cancer cells by targeting PETN[J]. Bioenc Rep, 2017,37(4):312-327.
[15]
Li Q S,Meng F Y,Zhao Y H,et al. Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells[J]. Bone Joint Res, 2017, 6(8):464-471.
[16]
Tatsumi S,Ishii K,Amizuka N,et al.Targeted ablation of osteocytes induces osteoporosis with defective mechanotransduction[J]. Cell Metab, 2007, 5(6):464-475.