Liu Hui, Peng Fengping, Liu Zhaoyun, Jiang Fengjuan, Li Lijuan, Gao Shan, Wang Guojin, Song Jia, Ruan Erbao, Shao Zonghong, Fu Rong
Department of Hematology, Tianjin Medical University General Hospital, Tianjin 300052, P.R. China.
Int J Oncol. 2017 Feb;50(2):631-639. doi: 10.3892/ijo.2016.3815. Epub 2016 Dec 22.
Cysteine-rich 61 (CYR61/CCN1), a secreted protein in bone marrow (BM) microenvironment, has diverse effects on many cellular activities such as growth and differentiation. However, the effect of CCN1 on osteoblasts (OBs) in myeloma bone disease remains unclear. In our study, the level of CCN1 in multiple myeloma (MM) patients was detected by ELISA and RT-PCR. The proliferation and differentiation of OBs from MM patients were observed after stimulated by CCN1 in vitro. The myeloma cells transduced with CYR61 gene (RPMI‑8226/CYR61) were injected in a mouse model to evaluate the efficacy of CCN1 in vivo and compare with zoledronic acid. The results showed that CYR61/CCN1 levels in BM supernatant and OBs both elevated significantly in all newly diagnosed MM patients, especially in patients without bone disease (P=0.001 and P<0.001). After 30 ng/l CCN1 stimulation for 24 h, the quantity and mineralization of OBs increased significantly in vitro (P=0.046 and 0.048). The transcription factors of Wnt pathway, runt-related transcription factor 2 (Runx2) and β-catenin were upregulated in OBs after CCN1 stimulation (P=0.012 and 0.011). After injection of RPMI‑8226 cells, bone lesions were observed obviously by microCT and histochemistry at 7 weeks. Radiographic analysis of the bones showed decreased resorption in CCN1 overexpression group and zoledronic acid group, while severe resorption in negative control. Furthermore, trabecular bone volume in CCN1 overexpression group (1.7539±0.16949) was significantly higher than zoledronic acid group (1.2839±0.077) (P=0.012). In conclusion, CCN1 can stimulate the proliferation and differentiation of OBs in vitro and contribute to bone remodeling in vivo in MBD.
富含半胱氨酸的61蛋白（CYR61/CCN1）是骨髓（BM）微环境中的一种分泌蛋白，对许多细胞活动如生长和分化具有多种作用。然而，CCN1在骨髓瘤骨病中对成骨细胞（OBs）的影响仍不清楚。在我们的研究中，通过酶联免疫吸附测定（ELISA）和逆转录聚合酶链反应（RT-PCR）检测了多发性骨髓瘤（MM）患者体内CCN1的水平。体外给予CCN1刺激后，观察了MM患者来源的OBs的增殖和分化情况。将转导了CYR61基因的骨髓瘤细胞（RPMI‑8226/CYR61）注射到小鼠模型中，以评估CCN1在体内的疗效，并与唑来膦酸进行比较。结果显示，所有新诊断的MM患者骨髓上清液和OBs中的CYR61/CCN1水平均显著升高，尤其是在无骨病的患者中（P = 0.001和P < 0.001）。在30 ng/l CCN1刺激24小时后，体外OBs的数量和矿化显著增加（P = 0.046和0.048）。CCN1刺激后，OBs中Wnt通路的转录因子，即 runt相关转录因子2（Runx2）和β-连环蛋白上调（P = 0.012和0.011）。注射RPMI‑8226细胞7周后，通过显微计算机断层扫描（microCT）和组织化学明显观察到骨病变。对骨骼的影像学分析显示，CCN1过表达组和唑来膦酸组的骨吸收减少，而阴性对照组骨吸收严重。此外，CCN1过表达组的骨小梁体积（1.7539±0.16949）显著高于唑来膦酸组（1.2839±0.077）（P = 0.012）。总之，CCN1在体外可刺激OBs的增殖和分化，并有助于骨髓瘤骨病（MBD）体内的骨重塑。
