J Feng1, J-X Wang, C-H Li. 1. Health Management Centre, Weifang People's Hospital, Weifang, China. licaihua12@163.com.
Abstract
OBJECTIVE: The aim of this study was to elucidate whether long non-coding RNA (lncRNA) GAS5 could target microRNA-498 to regulate RUNX2, thus alleviating the development of osteoporosis. PATIENTS AND METHODS: Human multipotential mesenchymal stem cells (hMSCs) were isolated from bone marrow of osteoporosis patients or healthy controls. Quantitative Real-time polymerase chain reaction (qRT-PCR) was performed to detect the expression levels of GAS5, microRNA-498 and RUNX2 in hMSCs of osteoporosis patients and controls, respectively. Meanwhile, the protein level of RUNX2 in hMSCs was detected by Western blot. Furthermore, alkaline phosphatase (ALP) activity assay and ALP staining were performed to evaluate the degree of osteogenic differentiation under the control of GAS5, microRNA-498 and RUNX2. RESULTS: MicroRNA-498 was highly expressed in hMSCs derived from osteoporosis patients, whereas GAS5 and RUNX2 were lowly expressed. GAS5 overexpression significantly increased ALP activity and promoted osteogenic differentiation of hMSCs derived from osteoporosis patients. Meanwhile, GAS5 significantly promoted osteogenic differentiation by mediating microRNA-498 expression to up-regulate RUNX2. Co-overexpression of GAS5 and microRNA-498 could remarkably reverse the increase of RUNX2 expression. Besides, RUNX2 overexpression markedly elevated ALP activity. CONCLUSIONS: LncRNA GAS5 is lowly expressed in patients with osteoporosis. Overexpression of GAS5 promotes osteogenic differentiation of hMSCs through regulating microRNA-498 to up-regulate RUNX2 expression, thus alleviating the development of osteoporosis.
OBJECTIVE: The aim of this study was to elucidate whether long non-coding RNA (lncRNA) GAS5 could target microRNA-498 to regulate RUNX2, thus alleviating the development of osteoporosis. PATIENTS AND METHODS: Human multipotential mesenchymal stem cells (hMSCs) were isolated from bone marrow of osteoporosispatients or healthy controls. Quantitative Real-time polymerase chain reaction (qRT-PCR) was performed to detect the expression levels of GAS5, microRNA-498 and RUNX2 in hMSCs of osteoporosispatients and controls, respectively. Meanwhile, the protein level of RUNX2 in hMSCs was detected by Western blot. Furthermore, alkaline phosphatase (ALP) activity assay and ALP staining were performed to evaluate the degree of osteogenic differentiation under the control of GAS5, microRNA-498 and RUNX2. RESULTS: MicroRNA-498 was highly expressed in hMSCs derived from osteoporosispatients, whereas GAS5 and RUNX2 were lowly expressed. GAS5 overexpression significantly increased ALP activity and promoted osteogenic differentiation of hMSCs derived from osteoporosispatients. Meanwhile, GAS5 significantly promoted osteogenic differentiation by mediating microRNA-498 expression to up-regulate RUNX2. Co-overexpression of GAS5 and microRNA-498 could remarkably reverse the increase of RUNX2 expression. Besides, RUNX2 overexpression markedly elevated ALP activity. CONCLUSIONS: LncRNA GAS5 is lowly expressed in patients with osteoporosis. Overexpression of GAS5 promotes osteogenic differentiation of hMSCs through regulating microRNA-498 to up-regulate RUNX2 expression, thus alleviating the development of osteoporosis.