Pro-apoptotic Effect of Recombinant Anti-HER2 Fusion Protein Scfv/Tbid on Osteosarcoma El0 Cells  

作　　者：Xiuchun Qiu Lequn Shan Sai Ma Zhengang Jl Tongtao Yang Hua Long Yanming Xu Yong Zhou Baoan Ma Angang Yang Qingyu Fan 

机构地区：[1]Department of Orthopaedics Surgery, Tangdu hospital, Xi ＇an 710038, China [2]Department of Prosthodontics, School of Stomatology, Fourth Military Medical University, Xi＇an 710038, China [3]Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi＇an 710038, China [4]Department of Immunology, Faculty of Preclinical Medicine,＂ Fourth Military Medical University, Xi＇an 710038, China

出　　处：《Journal of US-China Medical Science》2010年第4期14-19,共6页美中医学（英文版）

基　　金：The first two authors contributed equally to this study Acknowledgements This work was supported by grants from the National Natural Science Foundation of China （No. 30471988, 30873027, 30901784 and No.30330610） and Postdoctoral Science Foundation of China （No.2005038259）.

摘　　要：Objective： To construct the gene of a recombinant anti-HER2 fusion protein, ScFv/tBid, and investigate its pro-apopotic effect on osteosarcoma E10 cell line. Methods： Firsl, the expression of HER-2 in El0 cells was examined. Then the gene of recombinant tBid fusion protein was constructed by sequentially fusing the coding sequences of a signal peptide, a single-chain HER2 antibody, a PE translocation domain and tBid. The effect of the recombinant fusion protein on the morphology and growth status orE 10 cells was studied by immunofluorescent staining. Meanwhile, pro-apoptotic effect was observed by Annexin V-FITC staining and TUNEL staining. Results： The expression of HER-2 in El0 was higher than control group. The recombinant ScFv/tBid protein was expressed and cell shrinkage and nuclear condensation can be detected in E 10 cells transfected with pCMV-ScFv/tBid. The apoptosis rate in El0 cells transfected with pCMV-ScFv/tBid or pCMV was 16.1% and 4.5%, respectively. The pro-apoptotic effect of recombinant ScFv/tBid protein was further verified by TUNEL staining. Conclusion： The recombinant ScFv/tBid protein can induce apoptosis in HER2/neu -positive osteosarcoma.

关 键 词：TBID HER2 fusion protein osteosarcoma cell apoptosis. 

分 类 号：Q786[生物学—分子生物学] Q255