Walsh C A, Birch M A, Fraser W D, Lawton R, Dorgan J, Walsh S, Sansom D, Beresford J N, Gallagher J A
Department of Human Anatomy and Cell Biology, The University, Liverpool, United Kingdom.
J Bone Miner Res. 1995 Jan;10(1):17-25. doi: 10.1002/jbmr.5650100106.
We investigated the production of parathyroid hormone-related protein (PTHrP) by cells derived from explants of human bone. Using an immunoradiometric assay (IRMA), PTHrP was detected in conditioned medium from cultures of bone-derived cells from 6 of 7 patients investigated in this study. PTHrP mRNA was identified in human bone cells using reverse transcriptase-linked polymerase chain reaction (RT-PCR) and by Northern analysis. Transcripts for PTHrP were detected in a purified population of alkaline phosphatase positive cells isolated from human bone marrow cultures by flow cytometry, confirming the expression of PTHrP mRNA by cells of the osteoblastic lineage. Production of PTHrP was inhibited by 10(-6) M of the glucocorticoids, prednisolone and desacetylated deflazacort, in a dose-dependent manner. In addition, RT-PCR followed by Southern blot analysis detected a decrease in steady-state PTHrP mRNA in cultures of human bone-derived cells treated with 10(-6) M prednisolone.
我们研究了源自人骨外植体的细胞甲状旁腺激素相关蛋白（PTHrP）的产生情况。使用免疫放射分析（IRMA），在本研究中所调查的7例患者中，有6例的骨源性细胞培养条件培养基中检测到了PTHrP。利用逆转录聚合酶链反应（RT-PCR）和Northern分析在人骨细胞中鉴定出了PTHrP mRNA。通过流式细胞术从人骨髓培养物中分离出的碱性磷酸酶阳性细胞纯化群体中检测到了PTHrP转录本，证实了成骨细胞系细胞表达PTHrP mRNA。10⁻⁶ M的糖皮质激素泼尼松龙和去乙酰化地夫可特以剂量依赖方式抑制PTHrP的产生。此外，RT-PCR随后进行Southern印迹分析发现在用10⁻⁶ M泼尼松龙处理的人骨源性细胞培养物中，稳态PTHrP mRNA有所减少。
