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Targeted proapoptotic effect of human recombinant caspase6 fusion protein on osteosarcoma SOSP9607 cells
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Abstract:
To investigate the proapoptotic effect of Her 2targeted recombinant caspase6 fusion protein on osteosarcoma SOSP9607 cells. Methods: Recombinant immunocasp6 was generated by fusing the genes of a signal peptide, a singlechain Her 2 antibody (e23sFv), a PEA translocation domain (PEA aa253364), and an active caspase6. The constructed immunocasp6 gene（e23sFvPEⅡcasp6） was cloned into pCMV plasmid and transfected into SOSP9607 cells. The target gene expression and the morphology of the transfected cells were observed by fluorescence and electron microscopy, and the proapoptotic effect of the recombinant gene was analyzed by Annexin VFITC staining, flow cytometry, and MTT assay. Results： The e23sFvPEⅡcasp6 fusion protein was detected in the cytoplasm of transfected SOSP9607 cells. The transfected cells presented the typical characteristics of apoptosis as detected by electron microscopy (cytoplasm concentration, chromatin condensation, vacuolation). Annexin VFITC staining revealed that the percentage of apoptotic cells in the transfectants of immunocasp6 genes was 31.4％,compared with 6.0％ in the control cells. MTT assay showed that the proliferation of immunocasp6 genetransfected SOSP9607 cells was much lower than that of non or mocktransfected cells. Conclusion： Her 2 targeted recombinant immunocasp6 fusion protein can express in SOSP9607 cells and induce apoptosis of SOSP9607 cells.
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Clc Number:
R73-362
