Journal: Experimental and Therapeutic Medicine

Article Title: miR-150-5p inhibits osteogenic differentiation of fibroblasts in ankylosing spondylitis by targeting VDR

doi: 10.3892/etm.2022.11213

Figure Lengend Snippet: Downregulation of miR-150-5p in AS joint capsules and fibroblasts. (A) miR-150-5p expression in joint capsule tissues obtained from AS patients and non-AS controls (n=20/group) was determined by reverse transcription-quantitative PCR. (B) miR-150-5p expression in BMP-2 and TGF-β1-treated AS fibroblasts and untreated cells. Data are presented as the mean ± SD. ** P<0.01 vs. control. miR, microRNA; AS, ankylosing spondylitis; BMP, bone morphogenetic protein; TGF, transforming growth factor.

Article Snippet: For induction of osteogenic differentiation, cells were cultured at 37˚C in DMEM supplemented with 50 bone morphogenic protein 2 (BMP-2) and 10 ng/ml transforming growth factor-β1 (TGF-β1; both R&D Systems, Inc.) for 14 days ( ).

Techniques: Capsules, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Experimental and Therapeutic Medicine

Article Title: miR-150-5p inhibits osteogenic differentiation of fibroblasts in ankylosing spondylitis by targeting VDR

doi: 10.3892/etm.2022.11213

Figure Lengend Snippet: Overexpression of miR-150-5p suppresses osteogenic differentiation of AS fibroblasts. (A) Transfection efficiency. (B) mRNA expression of Col I, OPN and Runx2 determined by reverse transcription-quantitative PCR. (C) Protein levels of Col I, OPN and Runx2 determined by western blotting. (D) ALP activity. (E) Alizarin red staining for mineralization in BMP-2 and TGF-β1-induced AS fibroblasts transfected with miR-150-5p mimics or miR-NC or untreated cells (magnification, x100). Data are presented as the mean ± SD. ** P<0.01, *** P<0.001 vs. control; # P<0.05, ## P<0.01, ### P<0.001 vs. miR-NC. miR, microRNA; AS, ankylosing spondylitis; BMP, bone morphogenetic protein; TGF, transforming growth factor; Col I, collagen type I; OPN, osteopontin; ALP, alkaline phosphatase; NC, negative control.

Article Snippet: For induction of osteogenic differentiation, cells were cultured at 37˚C in DMEM supplemented with 50 bone morphogenic protein 2 (BMP-2) and 10 ng/ml transforming growth factor-β1 (TGF-β1; both R&D Systems, Inc.) for 14 days ( ).

Techniques: Over Expression, Transfection, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Activity Assay, Staining, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: miR-150-5p inhibits osteogenic differentiation of fibroblasts in ankylosing spondylitis by targeting VDR

doi: 10.3892/etm.2022.11213

Figure Lengend Snippet: miR-150-5p decreases VDR expression by targeting VDR 3'-UTR. (A) Schematic diagram of the predicted miR-150-5p binding sites to VDR. (B) Luciferase assay in AS fibroblasts transfected with pmirGLO-VDR-WT or -MUT reporters and miR-150-5p mimics or miR-NC. (C) Protein levels of VDR in AS fibroblasts transfected with miR-150-5p mimics or miR-NC. (D) mRNA expression of VDR in joint capsule tissues obtained from patients with AS and non-AS controls (n=20/group) determined by reverse transcription-quantitative PCR. (E) Spearman's correlation analysis of miR-150-5p and VDR mRNA expression in AS joint capsules. (F) VDR protein levels in BMP-2 and TGF-β1-treated AS fibroblasts or untreated cells. Data are presented as the mean ± SD. ** P<0.01, *** P<0.001 vs. miR-NC or control. miR, microRNA; AS, ankylosing spondylitis; BMP, bone morphogenetic protein; TGF, transforming growth factor; VDR, vitamin D receptor; UTR, untranslated region; WT, wild-type; MUT, mutant; NC, negative control.

Article Snippet: For induction of osteogenic differentiation, cells were cultured at 37˚C in DMEM supplemented with 50 bone morphogenic protein 2 (BMP-2) and 10 ng/ml transforming growth factor-β1 (TGF-β1; both R&D Systems, Inc.) for 14 days ( ).

Techniques: Expressing, Binding Assay, Luciferase, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Capsules, Mutagenesis, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: miR-150-5p inhibits osteogenic differentiation of fibroblasts in ankylosing spondylitis by targeting VDR

doi: 10.3892/etm.2022.11213

Figure Lengend Snippet: miR-150-5p regulates osteogenic differentiation in AS fibroblasts by downregulating VDR. (A) Transfection efficiency. (B) mRNA expression of Col I, OPN and Runx2. (C) Protein levels of Col I, OPN and Runx2. (D) ALP activity. (E) Alizarin red staining for mineralization in BMP-2 and TGF-β1-treated AS fibroblasts transfected with miR-NC or miR-150-5p mimics and with VDR (magnification, x100). Data are presented as the mean ± SD. * P<0.05, ** P<0.01, *** P<0.001 vs. Vector or miR-NC; # P<0.05, ## P<0.01, ### P<0.001 vs. miR-150-5p. miR, microRNA; AS, ankylosing spondylitis; BMP, bone morphogenetic protein; TGF, transforming growth factor; VDR, vitamin D receptor; UTR, untranslated region; Col I, collagen type I; OPN, osteopontin; ALP, alkaline phosphatase; NC, negative control.

Article Snippet: For induction of osteogenic differentiation, cells were cultured at 37˚C in DMEM supplemented with 50 bone morphogenic protein 2 (BMP-2) and 10 ng/ml transforming growth factor-β1 (TGF-β1; both R&D Systems, Inc.) for 14 days ( ).

Techniques: Transfection, Expressing, Activity Assay, Staining, Plasmid Preparation, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: miR-150-5p inhibits osteogenic differentiation of fibroblasts in ankylosing spondylitis by targeting VDR

doi: 10.3892/etm.2022.11213

Figure Lengend Snippet: Schematic diagram of the potential role of miR-150-5p/VDR pathway in BMP-2 + TGF-β1-induced AS osteogenic differentiation. Arrows indicate downregulation. miR, microRNA; AS, ankylosing spondylitis; BMP, bone morphogenetic protein; TGF, transforming growth factor; VDR, vitamin D receptor.

Article Snippet: For induction of osteogenic differentiation, cells were cultured at 37˚C in DMEM supplemented with 50 bone morphogenic protein 2 (BMP-2) and 10 ng/ml transforming growth factor-β1 (TGF-β1; both R&D Systems, Inc.) for 14 days ( ).

Techniques: