Kominsky Scott L, Abdelmagid Samir M, Doucet Michele, Brady Kelly, Weber Kristy L
Department of Orthopaedic Surgery, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Cancer Res. 2008 Mar 1;68(5):1261-6. doi: 10.1158/0008-5472.CAN-07-6122.
Approximately 30% of patients with renal cell carcinoma (RCC) develop bone metastasis, which is characterized by extensive osteolysis leading to severe bone pain and pathologic fracture. Although the mechanism of RCC-induced osteolysis is unknown, studies of bone metastasis have shown that tumor-induced changes in bone remodeling are likely mediated by alterations in the bone microenvironment. Here, we report the discovery of a novel osteoclast stimulatory factor secreted by RCC bone metastasis (RBM). Through microarray analysis, we found expression of the chemokine, macrophage inflammatory protein-1 delta (MIP-1 delta), to be increased in RBM versus patient-matched primary RCC tissues and confirmed this finding by quantitative reverse transcription-PCR (qRT-PCR) and ELISA (P < 0.05). Furthermore, MIP-1 delta expression in RBM tissues was significantly (P < 0.001) higher than in human bone marrow, suggesting a potential alteration of the bone microenvironment. The receptors for MIP-1 delta, CCR1 and CCR3, were expressed in both osteoclast precursors and mature, bone-resorbing osteoclasts as shown by qRT-PCR and Western analysis. In functional studies, MIP-1 delta stimulated chemotaxis of two osteoclast precursor cell types: murine bone marrow mononuclear cells (BM-MNC) and RAW 264.7 cells. Furthermore, MIP-1 delta treatment of murine calvaria caused increased bone resorption as determined by measurement of released calcium. Correspondingly, MIP-1 delta significantly enhanced osteoclast formation and activity in response to RANKL in both BM-MNC and RAW 264.7 cells. Taken together, these data suggest that MIP-1 delta expression is increased in RBM relative to RCC and bone marrow, and may promote RBM-induced osteolysis by stimulating the recruitment and differentiation of osteoclast precursors into mature osteoclasts.
约30%的肾细胞癌(RCC)患者会发生骨转移,其特征是广泛的骨质溶解,导致严重骨痛和病理性骨折。尽管RCC诱导骨质溶解的机制尚不清楚,但对骨转移的研究表明,肿瘤诱导的骨重塑变化可能是由骨微环境的改变介导的。在此,我们报告发现了一种由RCC骨转移(RBM)分泌的新型破骨细胞刺激因子。通过微阵列分析,我们发现趋化因子巨噬细胞炎性蛋白-1δ(MIP-1δ)在RBM中的表达相对于患者匹配的原发性RCC组织有所增加,并通过定量逆转录PCR(qRT-PCR)和酶联免疫吸附测定(ELISA)证实了这一发现(P<0.05)。此外,RBM组织中MIP-1δ的表达显著高于人骨髓(P<0.001),提示骨微环境可能发生了改变。如qRT-PCR和蛋白质免疫印迹分析所示,MIP-1δ的受体CCR1和CCR3在破骨细胞前体以及成熟的、具有骨吸收功能的破骨细胞中均有表达。在功能研究中,MIP-1δ刺激了两种破骨细胞前体细胞类型的趋化作用:小鼠骨髓单个核细胞(BM-MNC)和RAW 264.7细胞。此外,通过测量释放的钙来确定,MIP-1δ处理小鼠颅骨后导致骨吸收增加。相应地,MIP-1δ在BM-MNC和RAW 264.7细胞中均显著增强了对核因子κB受体活化因子配体(RANKL)的反应,促进破骨细胞的形成和活性。综上所述,这些数据表明,相对于RCC和骨髓,MIP-1δ在RBM中的表达增加,并且可能通过刺激破骨细胞前体募集和分化为成熟破骨细胞来促进RBM诱导的骨质溶解。