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Crosstalk between the COX2‐PGE2‐EP4 signaling pathway and primary cilia in osteoblasts after mechanical stimulation

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Primary cilia have been found to function as mechanosensors in low‐magnitude high‐frequency vibration (LMHFV)‐induced osteogenesis. The PGE2 also regulates bone homeostasis and mechanical osteogenesis through its receptor EP4 signaling, but… Click to show full abstract

Primary cilia have been found to function as mechanosensors in low‐magnitude high‐frequency vibration (LMHFV)‐induced osteogenesis. The PGE2 also regulates bone homeostasis and mechanical osteogenesis through its receptor EP4 signaling, but its involvement in LMHFV‐induced or in primary cilia‐induced osteogenesis has not been investigated. We hypothesized that LMHFV stimulates osteoblast (OB) differentiation by activating the COX2‐PGE2‐EP pathway in a manner dependent on primary cilia and that primary cilia are also affected by the PGE2 pathway. In this study, through western blot analysis, RNA interference, enzyme‐linked immunosorbent assay, real‐time quantitative polymerase chain reaction, and cytochemical staining, we observed that COX2, mPGES‐1, and PGE2 levels were markedly elevated in cells treated with LMHFV and were greatly decreased in LMHFV‐treated cells following IFT88 silencing. EP4 expression was significantly increased in OBs following LMHFV treatment, but IFT88 silencing significantly blocked this increase. EP4 localized to the bases of primary cilia. LMHFV reduced the length and abundance of primary cilia, but the cells could self‐repair their primary cilia after mechanical damage. EP4 antagonism significantly blocked the LMHFV‐induced increase in IFT88 expression and blocked the recovery of primary cilia length and the proportion of cells with primary cilia. In addition, COX2 or EP4 antagonism disrupted LMHFV‐induced osteogenesis. These results demonstrate the integration of and crosstalk between primary cilia and the COX2‐PGE2‐EP4 signaling pathway under mechanical stimulation.

Keywords: cox2 pge2; ep4 signaling; lmhfv; primary cilia; ep4

Journal Title: Journal of Cellular Physiology
Year Published: 2020

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