SDF-1CXCR4在BMP9介导C2C12细胞成骨分化过程中的.PDFVIP

中国细胞生物学学报 Chinese Journal of Cell Biology 2013, 35(2): 154– 162 http ://www.cj SDF-1/CXCR4在BMP9介导C2C12细胞成骨分化 过程中的作用研究 刘 晨 杨丹丹 白慧丽 李宝林 何 方 张汝益 严树涓 施 琼* (重庆医科大学检验医学院临床检验诊断学教育部重点实验室, 重庆市重点实验室, 重庆400016) 摘要 为了观察SDF-1/CXCR4信号轴在BMP9促C2C12细胞成骨分化过程中的作用, 通过重 组腺病毒过表达BMP9, 检测对C2C12细胞中SDF-1及受体CXCR4 mRNA和蛋白表达水平的影响; 同时利用重组腺病毒或中和抗体干扰SDF-1/CXCR4, 与BMP9先后作用于C2C12细胞, 通过定量测 定碱性磷酸酶(ALP) 、染色测定ALP表达、免疫细胞化学测定骨钙蛋白(OCN)表达、茜素红S染色 测定钙盐沉积、Real-time PCR检测成骨相关转录因子Runx2和Osx 的表达、Western blot检测成骨 分化信号通路MAPK和Smad的变化。结果显示, BMP9能明显抑制C2C 12 细胞中SDF- 1、CXCR4 的表达(P0.01), 且具有剂量和时间依赖性; 预先干扰SDF-1/CXCR4 能明显影响由BMP9介导的早、 中期成骨标志物ALP 、OCN及早期转录因子Runx2 、Osx的表达(P0.01)和MAPK 、Smad信号通路 相关蛋白的变化(P0.05); 外源性SDF-1并不能影响晚期成骨标志物钙盐沉积。提示SDF-1/CXCR4 信号轴在由BMP9介导的C2C12细胞成骨分化早、中期过程中发挥重要作用。 关键词 SDF-1; CXCR4; BMP9; 成骨分化; C2C12细胞 The Role for SDF-1/CXCR4 Signal Axis in BMP9-induced Osteogenic Differentiation of C2C12 Cells Liu Chen, Yang Dandan, Bai Huili, Li Baolin, He Fang, Zhang Ruyi, Yan Shujuan, Shi Qiong* (Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Faculty of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China) Abstract To elucidate the role of SDF-1/CXCR4 signal axis during BMP9-incuced osteogenic differen- tiation in C2C12 cells, BMP9 was introduced by using recombinant adenoviruses assay. The mRNA and protein expression levels of SDF-1 and CXCR4 induced by Ad-BMP9 were detected in C2C12 cells. At the same time, the recombinant adenovirus and neutralizing antibody were used to perturbing the SDF-1/CXCR4 signal axis in C2C12 cells before or after the addition of BMP9. The alkaline phosphatase (ALP) quantitative assay and fast blue RR salt staining were used to determine the express