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Construction of recombinant replication defective adenoviral vectors carrying attenuated Shigalike toxinⅠmutant gene and their therapeutic effects on breast cancer
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Abstract:
Objective:To construct recombinant replication defective adenoviral vectors encoding 1/100 and 1/1000 attenuated Shigalike toxinⅠ(Stx1) gene, and to study their therapeutic effects against breast cancer T47D cells in vivo.Methods: Genes encoding 1/100 or 1/1000 attenuated Stx1 were amplified by overlapping PCR and were cloned into T vectors. The inserted gene was verified by nucleotide sequencing. Replication defective adenoviral vector AdvStx1R170L containing 1/1000 attenuated Stx1 mutant gene was generated by AdMAX Adenoviral Vector System. Nude mouse models bearing human breast cancer T47D cells were established, and the tumor inhibitory effect of AdvStx1R170L was studied by intratumor injection of the recombinant adenovirus. Results: Vectors carrying 1/100 or 1/1000 attenuated Stx1 gene were successfully constructed and were verified by nucleotide sequencing. Recombinant replication defective adenoviral vector AdvStx1R170L containing 1/1000 attenuated Shigalike toxinⅠgene was constructed. In vivo study showed that AdvStx1R170L significantly inhibited the growth of implanted T47D tumor in the nude mice compared with AdvGFP and PBS group（P<0.05）. Conclusion: Recombinant replication defective adenoviral vector AdvStx1R170L encoding 1/1000 attenuated Shigalike toxin Ⅰgene has been successfully constructed, and it can effectively inhibit the growth of implanted T47D tumor in nude mice, without obvious toxicity.
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Project Supported:
Supported by Science and Technology Fund of Tianjin Health Bureau（No.05KY32）; the Doctoral Fund of Affiliated Tumor Hospital of Tianjin Medical University（No.07B02）
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