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Nuoya Cai,Chao Li,Fuke Wang 연세대학교의과대학 2019 Yonsei medical journal Vol.60 No.8
Purpose: This study aimed to explore the effects and mechanisms of long non-coding RNA (lncRNA) anti-differentiation noncodingRNA (ANCR) on the osteogenesis of osteoblast cells in postmenopausal osteoporosis (PMOP). Materials and Methods: Mice models of PMOP were established. ANCR expression and intracellular calcium ions were detectedby quantitative real-time polymerase chain reaction (qRT-PCR) and laser confocal microscopy, respectively. ANCR was silencedin osteoblast cells from PMOP mice by the transfection of siRNA-ANCR (si-ANCR). The proliferation and apoptosis of osteoblast cellswas analyzed by MTT and flow cytometry, respectively. Alkaline phosphatase (ALP) activity and calcium nodules were examinedby ALP and alizarin red staining assay, respectively. The expression of enhancer of zeste homolog 2 (EZH2), runt related transcriptionfactor 2 (RUNX2), and OSTERIX was detected by qRT-PCR and Western blot. Furthermore, an osteogenesis model was constructedin mice, and osteoid formation was observed by hematoxylin-eosin (HE) staining. The interaction between lncRNA-ANCRand EZH2 was further identified by RNA pull-down assay. Results: ANCR expression and intracellular calcium ions were increased in PMOP mice. Si-ANCR significantly increased the proliferation,ALP activity, calcium deposition of osteoblast cells and decreased apoptosis. ANCR and EZH2 were down-regulated bysi-ANCR, while RUNX2 and OSTERIX were upregulated. Si-ANCR also promoted osteoid formation in mice treated with hydroxyapatite-tricalcium phosphate. In addition, ANCR specifically bound to EZH2. Conclusion: Silencing ANCR promotes the osteogenesis of PMOP osteoblast cells. The specific binding of ANCR with EZH2 suppressedRUNX2, thereby inhibiting osteogenesis.