Liu Ji-Zhong, Ji Zong-Ling, Hu Yun-Yu, Chen Su-Min, Zhu Bang-Fu, Yang Tong-Tao
The Fourth Military Medical University, Department of Orthopedics, Xijing Hospital, China.
Sheng Wu Gong Cheng Xue Bao. 2003 Jan;19(1):35-40.
Using the isolated total RNA from osteosacoma cell line MG63, the cDNA encoding human OPG was amplified by RT-PCR. A recombinant adenoviral vector carrying cDNA of OPG was constructed and OPG expression in mouse myoblast C2C12 cells was confirmed by Western blot and ELISA. The secreted expression of OPG protein persisted more than 6 weeks in vitro, and the growth of C2C12 cells infected by recombinant adenoviral were in good state. Osteoclasts derived from mouse bone marrow cells infected with recombinant adenoviral made less number of TRAP positive cells and resorption pits formed on dentine slices.
利用骨肉瘤细胞系MG63中分离出的总RNA,通过逆转录聚合酶链反应(RT-PCR)扩增编码人骨保护素(OPG)的cDNA。构建携带OPG cDNA的重组腺病毒载体,并通过蛋白质免疫印迹法(Western blot)和酶联免疫吸附测定(ELISA)证实其在小鼠成肌细胞C2C12中的表达。OPG蛋白的分泌表达在体外持续超过6周,并且感染重组腺病毒的C2C12细胞生长状态良好。感染重组腺病毒的小鼠骨髓细胞来源的破骨细胞形成的抗酒石酸酸性磷酸酶(TRAP)阳性细胞数量减少,在牙本质切片上形成的吸收陷窝也减少。