MicroRNA-200a-3p accelerates the progression of osteoporosis by targeting glutaminase to inhibit osteogenic differentiation of bone marrow mesenchymal stem cells.

To uncover the role of microRNA-200a-3p in regulating osteogenic differentiation of MSCs via targeting glutaminase, thus influencing the progression of OP. Serum level of microRNA-200a-3p in OP patients and healthy controls was determined by qRT-PCR. MicroRNA-200a-3p level in MSCs undergoing osteogenic differentiation for different days was examined as well. ALP activity, calcification nodules and relative levels of Bglap, Runx2 and OPN in MSCs overexpressing microRNA-200a-3p undergoing osteogenic differentiation were detected. Relative l-glutaminase uptake in MSCs undergoing osteogenic differentiation for different days was determined. After transfection of si-GLS in MSCs undergoing osteogenic differentiation, l-glutaminase uptake, ALP activity and relative levels of Bglap, Runx2 and OPN were detected. The potential binding relationship between microRNA-200a-3p and GLS was tested by dual-luciferase reporter gene assay. Finally, rescue experiments were conducted to elucidate the role of microRNA-200a-3p/GLS in osteogenic differentiation of MSCs. MicroRNA-200a-3p level was higher in serum of OP patients relative to controls. Its level in MSCs gradually decreased with the prolongation of osteogenic differentiation. Overexpression of microRNA-200a-3p reduced cell viability, ALP activity, number and volume of calcification nodule. The mRNA levels of Bglap, Runx2 and OPN were downregulated by overexpressed microRNA-200a-3p. The cell viability, ALP activity, number and volume of calcification nodule were reduced when microRNA-200a-3p was knocked down. The mRNA levels of Bglap, Runx2 and OPN were upregulated when transfected microRNA-200a-3p inhibitor. l-glutaminase uptake increased with the prolongation of osteogenic differentiation in MSCs. Knockdown of GLS attenuated l-glutaminase uptake and ALP activity, as well as downregulated Bglap, Runx2 and OPN. Besides, GLS was verified to directly bind to microRNA-200a-3p. GLS overexpression reversed the inhibitory effects of overexpressed microRNA-200a-3p on osteogenic differentiation of MSCs. MicroRNA-200a-3p suppresses osteogenic differentiation of MSCs via targeting glutaminase, thereafter accelerating the progression of OP.

Lv R ，Pan X ，Song L ，Sun Q ，Guo C ，Zou S ，Zhou Q ... -  《-》

MicroRNA-579-3p promotes the progression of osteoporosis by inhibiting osteogenic differentiation of mesenchymal stem cells through regulating Sirt1.

Luo B ，Yang JF ，Wang YH ，Qu GB ，Hao PD ，Zeng ZJ ，Yuan J ，Yang R ，Yuan Y ... -  《-》

MiRNA-19a-3p alleviates the progression of osteoporosis by targeting HDAC4 to promote the osteogenic differentiation of hMSCs.

To clarify the function of microRNA-19a-3p (miRNA-19a-3p) in the osteogenic differentiation of human-derived mesenchymal stem cells (hMSCs) and the potential mechanism. Serum levels of miRNA-19a-3p, RUNX2 and OCN in osteoporosis patients and controls were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Alkaline phosphatase (ALP) content and calcification ability during the process of osteogenic differentiation were examined by ALP staining and alizarin red staining, respectively. After altering miRNA-19a-3p level by transfection of miRNA-19a-3p mimic or inhibitor, we detected relative levels of miRNA-19a-3p, RUNX2 and OCN in hMSCs by qRT-PCR. The binding relationship between miRNA-19a-3p and HDAC4 was predicted by TargetScan and further verified by dual-luciferase reporter gene assay. Relative expression of HDAC4 was detected by Western blot and qRT-PCR in hMSCs transfected with miRNA-19a-3p mimic or inhibitor. Regulatory effects of miRNA-19a-3p/HDAC4 axis on osteogenic differentiation of hMSCs were evaluated. MiRNA-19a-3p was downregulated in osteoporosis patients. Its level gradually increased in hMSCs with the prolongation of osteogenic differentiation. Overexpression of miRNA-19a-3p upregulated levels of RUNX2 and OCN, and enhanced ALP activity. Knockdown of miRNA-19a-3p obtained the opposite trends. Dual-luciferase reporter gene assay verified that miRNA-19a-3p could target to 3'UTR of HDAC4. Protein level of HDAC4 was negatively regulated by miRNA-19a-3p in hMSCs. More importantly, co-overexpression of miRNA-19a-3p and HDAC4 could reverse the regulatory effects of miRNA-19a-3p on enhancing ALP activity and upregulating RUNX2 and OCN. MiRNA-19a-3p promotes the osteogenic differentiation of hMSCs by inhibiting HDAC4 expression, thus alleviating the progression of osteoporosis.

Chen R ，Qiu H ，Tong Y ，Liao F ，Hu X ，Qiu Y ，Liao Y ... -  《-》

MiRNA-27a-3p promotes osteogenic differentiation of human mesenchymal stem cells through targeting ATF3.

Fu YC ，Zhao SR ，Zhu BH ，Guo SS ，Wang XX ... -  《-》

miR-206 inhibits osteogenic differentiation of bone marrow mesenchymal stem cells by targetting glutaminase.

Osteoblast-mediated bone formation is a complex process involving various pathways and regulatory factors, including cytokines, growth factors, and hormones. Investigating the regulatory mechanisms behind osteoblast differentiation is important for bone regeneration therapy. miRNAs are known as important regulators, not only in a variety of cellular processes, but also in the pathogenesis of bone diseases. In the present study, we investigated the potential roles of miR-206 during osteoblast differentiation. We report that miR-206 expression was significantly down-regulated in human bone marrow mesenchymal stem cells (BMSCs) at days 7 and 14 during osteogenic induction. Furthermore, miR-206 overexpressing BMSCs showed attenuated alkaline phosphatase (ALP) activity, Alizarin Red staining, and osteocalcin secretion. The mRNA levels of osteogenic markers, Runx2 and Osteopontin (OPN), were significantly down-regulated in miR-206 overexpressing BMSCs. We observed that significantly increased glutamine uptake at days 7 and 14 during the osteogenic induction and inhibition of glutamine metabolism by knocking down glutaminase (GLS)-suppressed osteogenic differentiation of BMSCs. Here, we discover that miR-206 could directly bind to the 3'-UTR region of GLS mRNA, resulting in suppressed GLS expression and glutamine metabolism. Finally, restoration of GLS in miR-206 overexpressing BMSCs led to recovery of glutamine metabolism and osteogenic differentiation. In summary, these results reveal a new insight into the mechanisms of the miR-206-mediated osteogenesis through regulating glutamine metabolism. Our study may contribute to the development of therapeutic agents against bone diseases.

Chen Y ，Yang YR ，Fan XL ，Lin P ，Yang H ，Chen XZ ，Xu XD ... -  《-》