The Targeted Pro-apoptotic Effect of scFv-mediated Delivery of Caspase-6 on HER2-positive Osteosarcoma Cells  

作　　者：SHAN Le-qun ZHOU Ben-gen QIU Xiu-chun XU Yan-ming WANG Yu-cai MA Bao-an ZHOU Yong FAN Qing-yu 

机构地区：[1]Department of Orthopaedics Surgery, Tangdu Hospital, the Fourth Military Medical University, Xi＇an 710038 [2]State Key Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, the Fourth Military Medical University, Xi＇an 710032

出　　处：《Journal of US-China Medical Science》2008年第1期1-6,共6页美中医学（英文版）

基　　金：Acknowledgements： This work was supported by grants from the National Basic Research Program of China （No. 2004CB518805 and （No. 2006AA02A255）, the National Natural Science Foundation of China （No. 30471988 and No. 30330610） and Postdoctoral Science Foundation of China （No.2005038259）.

摘　　要：Objective To investigate the pro-apoptotic effect of scFv-mediated delivery of caspase-6 fusion protein on osteosarcoma E 10 cells. Methods The expression of human epidermal growth factor receptor-2 （Her-2） in osteosarcoma cell line E10 was examined by flow cytometry analysis. ScFv-mediated delivery of caspase-6 was generated by sequential fusion of the genes of a signal peptide, a single-chain Her-2 antibody （e23sFv）, a PEA translocation domain （PEA aa253-264）, and an recombinant caspase-6. The pCMV-immunocaspase-6 was produced by cloning above mentioned fusion genes into pCMV plasmid, and then pCMV-immunocaspase-6 plasmid mixed cationic liposome was transfected into E10 cells. The expression of fusion gene on transfected E10 cells was detected by immunocytochemical staining, The pro-apoptotic effects of fusion gene expression on morphology and growth status of E10 cells were observed by trypan blue cell staining, electron microscopy and MTT assay. Results The fusion protein was detected in the cytoplasm of the transfected E10 cells. These cells presented typical characteristics of apoptosis as detected by electron microscopy. MTT assay revealed that the proliferation of transfected E10 cells was suppressed than that of non- or mock-transfected cells （P〈0.01）. Conclusion ScFv-mediated delivery of caspase-6 fusion protein can induce apoptosis of Her-2 positive osteosarcoma E10 cells, suggesting the potential of this strategy for the treatment of human high-grade osteosarcoma.

关 键 词：OSTEOSARCOMA gene therapy HER-2/NEU PEA CASPASE-6 fusion protein apoptosis 

分 类 号：R738.1[医药卫生—肿瘤]