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Targeted Tumor Supprssion by a Secreted Fusion Protein Consisting of Anti-erbB2 Antibody and Reversed caspase-3 to Three Kinds of Cells
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Abstract:
Objiective：To investigate the targeted killing effect to ErbB2 antigen positive cells due to the expression of a secreted fusion protein consisting of anti-erB2 single chain antibody and reversed caspase-3 protein. Methods：pCMV-e23scFv-PeⅡ-revcasp-3 was constructed by sub cloning reversed caspase-3 gene to downstream of anti-erbB2 antibody and PE40 domain Ⅱ genes in recombinant pCMV vector and transfect Jurkat cells. The cell lines secreted expressing fusion protein stable were selected. Fusion protein in mediate was detected by ELISA. Then such mediate was used to culture Hela, SKBr3 and SKOV3 cells respectively and their survivals were compared through MTT. Finally Jurkat- pCMV-e23scFv-PEⅡ-revcasp-3 cells were administrated to BALB/c nude mice bearing SKBr3 tumor through their tail veins. Results： Fusion protein could be expressed by Jurkat cells stably and kill SKBr3 and SKOV3 cells which express ErbB2 antigen but had no influence on Hela which does not expressed ErbB2 antigen. Administrating Jurkat- pCMV-e23scFv-PEⅡ-revcasp-3 cells could inhibit SKBr3 tumor in vivo. Conclusions: Secreted expression of the fusion protein consisting of anti-erbB2 antibody and reversed caspase-3 could targetedly induce ErbB2 antigen positive cells to death.
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