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Dexamethasone Regulates EphA5 a Potential Inhibitory Factor with Osteogenic Capability of Human Bone Marrow Stromal Cells

机译:地塞米松调节EphA5一种具有潜在成骨能力的人骨髓基质细胞成骨能力的抑制因子。

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摘要

We previously demonstrated the importance of quality management procedures for the handling of human bone marrow stromal cells (hBMSCs) and provided evidence for the existence of osteogenic inhibitor molecules in BMSCs. One candidate inhibitor is the ephrin type-A receptor 5 (EphA5), which is expressed in hBMSCs and upregulated during long-term culture. In this study, forced expression of EphA5 diminished the expression of osteoblast phenotypic markers. Downregulation of endogenous EphA5 by dexamethasone treatment promoted osteoblast marker expression. EphA5 could be involved in the normal growth regulation of BMSCs and could be a potential marker for replicative senescence. Although Eph forward signaling stimulated by ephrin-B-Fc promoted the expression of ALP mRNA in BMSCs, exogenous addition of EphA5-Fc did not affect the ALP level. The mechanism underlying the silencing of EphA5 in early cultures remains unclear. EphA5 promoter was barely methylated in hBMSCs while histone deacetylation could partially suppress EphA5 expression in early-passage cultures. In repeatedly passaged cultures, the upregulation of EphA5 independent of methylation could competitively inhibit osteogenic signal transduction pathways such as EphB forward signaling. Elucidation of the potential inhibitory function of EphA5 in hBMSCs may provide an alternative approach for lineage differentiation in cell therapy strategies and regenerative medicine.
机译:我们先前证明了质量管理程序对于处理人骨髓基质细胞(hBMSCs)的重要性,并为BMSCs中存在成骨抑制剂分子提供了证据。一种候选抑制剂是ephrin A型受体5(EphA5),它在hBMSC中表达并在长期培养过程中被上调。在这项研究中,EphA5的强制表达减少了成骨细胞表型标记的表达。地塞米松处理下调内源性EphA5可促进成骨细胞标志物的表达。 EphA5可能参与BMSCs的正常生长调节,并可能是复制性衰老的潜在标志。尽管ephrin-B-Fc刺激的Eph正向信号传导促进了BMSCs中ALP mRNA的表达,但外源添加EphA5-Fc不会影响ALP水平。早期培养中EphA5沉默的潜在机制尚不清楚。 EphA5启动子在hBMSC中几乎没有被甲基化,而组蛋白去乙酰化可以部分抑制早期传代培养物中EphA5的表达。在反复传代的培养物中,独立于甲基化的EphA5的上调可以竞争性地抑制成骨信号转导途径,例如EphB正向信号传导。阐明hBMSCs中EphA5的潜在抑制功能可能为细胞治疗策略和再生医学中的谱系分化提供另一种方法。

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