title: MiR-27a targets sFRP1 in hFOB cells to regulate proliferation, apoptosis and differentiation.
abstract: MicroRNAs (miRNAs) play a key role in the regulation of almost all the physiological and pathological processes, including bone metabolism. Recent studies have suggested that miR-27 might play a key role in osteoblast differentiation and bone formation. Increasing evidence indicates that the canonical Wnt signaling pathway contributes to different stages of bone formation. In this study, we identify miR-27a can promote osteoblast differentiation by repressing a new target, secreted frizzled-related proteins 1 (sFRP1) expression at the transcriptional level. Here, 21 candidate targets of miR-27a involved in canonical Wnt/β-catenin signaling were predicted, and a significant decrease in sFRP1 luciferase activity was observed both in 293T and MG63 cells co-transfected with the matched luciferase reporter constructs and miR-27a mimic. Furthermore, the presence of exogenous miR-27a significantly decreased sFRP1 mRNA and protein expression in hFOB1.19 cells during both proliferation and osteogenic differentiation. The over-expression of miR-27a or knockdown sFRP1 significantly increased the percentage of apoptotic hFOBs, the percentage of cells in the G2-M phase of the cell cycle and the expression of key osteoblastic markers, including ALP, SPP1, RUNX2 and ALP activity. Over-expression of miR-27a or knockdown of endogenous sFRP1 led to an accumulation of β-catenin in hFOBs. In the present study, we demonstrate that miR-27a induced gene silencing effect is a vital mechanism contributing to bone metabolism in hFOB cells in vitro, which is partly affected by the post-transcriptional regulation of sFRP1, during osteoblast proliferation, apoptosis and differentiation.
authors.affiliation: Department of Rheumatology, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China.
authors.forename: Donggeng
authors.initials: D
authors.lastname: Guo
authors.affiliation: Department of Rheumatology, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China.
authors.forename: Qiuxia
authors.initials: Q
authors.lastname: Li
authors.affiliation: Department of Rheumatology, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Lv
authors.affiliation: Department of Rheumatology, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China.
authors.forename: Qiujing
authors.initials: Q
authors.lastname: Wei
authors.affiliation: Department of Rheumatology, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China.
authors.forename: Shuangyan
authors.initials: S
authors.lastname: Cao
authors.affiliation: Department of Rheumatology, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China.
authors.forename: Jieruo
authors.initials: J
authors.lastname: Gu
mesh_terms: D020413:3' Untranslated Regions; D000469:Alkaline Phosphatase / Q000378:metabolism; D017209:Apoptosis; D001665:Binding Sites; D001842:Bone and Bones / Q000502:physiology; D002453:Cell Cycle; D002454:Cell Differentiation; D049109:Cell Proliferation; D019295:Computational Biology; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D057809:HEK293 Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D035683:MicroRNAs / Q000378:metabolism*; D009841:Oligonucleotides / Q000235:genetics; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D010012:Osteogenesis; D011506:Proteins / Q000378:metabolism*; D015398:Signal Transduction; D014162:Transfection; D051176:beta Catenin / Q000378:metabolism; D000099274:Frizzled-Related Proteins
chemical_list: D020413:3' Untranslated Regions; D000469:Alkaline Phosphatase; D047908:Intracellular Signaling Peptides and Proteins; D035683:MicroRNAs; D009841:Oligonucleotides; D011506:Proteins; D051176:beta Catenin; D000099274:Frizzled-Related Proteins; C495270:CTNNB1 protein, human; C531388:MIRN27 microRNA, human