Regulated expression of the diphtheria toxin A chain by a tumor-specific chimeric transcription factor results in selective toxicity for alveolar rhabdomyosarcoma cells
| Autor: | Edward J. Dunphy, Rebecca A. Redman, Timothy P. Cripe, Frederic G. Barr, Edmond S. Massuda, Jennifer J. Schreiber, Ian H. Maxwell, Lauren E. Nauta |
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| Rok vydání: | 1997 |
| Předmět: |
Recombinant Fusion Proteins
Muscle Proteins Nerve Tissue Proteins Biology Plasmid Gene expression Humans Cytotoxic T cell Diphtheria Toxin Rhabdomyosarcoma Alveolar Homeodomain Proteins Diphtheria toxin Regulation of gene expression Reporter gene Multidisciplinary Forkhead Box Protein O1 Gene Transfer Techniques PAX7 Transcription Factor Forkhead Transcription Factors Genetic Therapy Transfection Biological Sciences Fusion protein Molecular biology DNA-Binding Proteins Gene Expression Regulation Neoplastic HeLa Cells Transcription Factors |
| Zdroj: | Proceedings of the National Academy of Sciences. 94:14701-14706 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.94.26.14701 |
| Popis: | Alveolar rhabdomyosarcoma (ARMS) cells often harbor one of two unique chromosomal translocations, either t(2;13)(q35;q14) or t(1;13)(p36;q14). The chimeric proteins expressed from these rearrangements, PAX3-FKHR and PAX7-FKHR, respectively, are potent transcriptional activators. In an effort to exploit these unique cancer-specific molecules to achieve ARMS-specific expression of therapeutic genes, we have studied the expression of a minimal promoter linked to six copies of a PAX3 DNA binding site, prs-9. In transient transfections, expression of the prs-9-regulated reporter genes was ≈250-fold higher than expression of genes lacking the prs-9 sequences in cell lines derived from ARMS, but remained at or below baseline levels in other cells. High expression of these prs-9-regulated genes was also observed in a cancer cell line that lacks t(2;13) but was stably transfected with a plasmid expressing PAX3-FKHR. Transfection of a plasmid containing the diphtheria toxin A chain gene regulated by prs-9 sequences (pA3–6PED) was selectively cytotoxic for PAX3-FKHR-expressing cells. This was shown by inhibition of gene expression from cotransfected plasmids and by direct cytotoxicity after transfected cells were isolated by cell sorting. Gene transfer of pA3–6PED may thus be useful as a cancer-specific treatment strategy for t(2;13)- or t(1;13)-positive ARMS. Furthermore, gene transfer of fusion protein-regulated toxin genes might also be applied to the treatment of other cancers that harbor cancer-specific chromosomal translocations involving transcription factors. |
| Databáze: | OpenAIRE |
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