Adenoviral Bid Overexpression Induces Caspase-dependent Cleavage of Truncated Bid and p53-independent Apoptosis in Human Non-small Cell Lung Cancers
| Autor: | Laurie B. Owen-Schaub, Takuya Fukazawa, Barbara N. Walter |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Programmed cell death
Lung Neoplasms Time Factors Cell Survival Truncated BID Blotting Western Genetic Vectors Apoptosis DNA Fragmentation urologic and male genital diseases Biochemistry Adenoviridae Membrane Potentials Carcinoma Non-Small-Cell Lung Tumor Cells Cultured medicine Humans heterocyclic compounds Viability assay neoplasms Molecular Biology Caspase Cisplatin biology Gene Transfer Techniques Cell Biology Flow Cytometry Molecular biology digestive system diseases Mitochondria Cell culture Caspases biology.protein DNA fragmentation Tumor Suppressor Protein p53 biological phenomena cell phenomena and immunity Carrier Proteins BH3 Interacting Domain Death Agonist Protein DNA Damage medicine.drug |
| Zdroj: | Journal of Biological Chemistry. 278:25428-25434 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m302058200 |
| Popis: | Proapoptotic gene transfer to promote death or to augment killing by DNA-damaging agents represents a promising strategy for cancer therapy. We have constructed an adenoviral Tet-Off trade mark vector with tightly controlled expression of Bid (Ad-Bid) (Clontech, Palo Alto, CA). Using the non-small cell lung cancer cell lines H460, H358, and A549, low dose Ad-Bid was shown to induce high levels of full-length Bid as well as caspase-3 and -9 activity. Although only a small fraction of Bid was processed to truncated Bid (a step inhibited by benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone), Ad-Bid gene transfer resulted in mitochondrial changes consistent with apoptosis (mitochondrial depolarization, cytochrome c release), DNA fragmentation, and a dramatic loss of cell viability. The proapoptotic effects of Ad-Bid were independent of p53 status and were augmented markedly by caspase-8 activators such as the DNA-damaging agent cisplatin. When Ad-Bid and cisplatin were used together, chemosensitivity was restored in p53-null H358 cells, increasing death from 35% following treatment with cisplatin and Ad-LacZ to >90% death with Ad-Bid and cisplatin (Ad-Bid alone induced 50% cell death under these conditions). Ad-Bid can induce apoptosis in malignant cells and enhance chemosensitivity in the absence of p53, suggesting this approach as a potential cancer therapy. |
| Databáze: | OpenAIRE |
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