Gene therapy of pancreatic cancer with green fluorescent protein and tumor necrosis factor-related apoptosis-inducing ligand fusion gene expression driven by a human telomerase reverse transcriptase promoter.

Abstract

BACKGROUND:

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in malignant cells but not in normal cells. Ad/g-TRAIL, an adenoviral vector in which expression of green fluorescent protein (GFP) and TRAIL is driven by a human telomerase reverse transcriptase promoter, has shown promise as a targeted antitumor agent.

METHODS:

To investigate the effects of TRAIL gene therapy on pancreatic cancer, BxPC-3, MIA-PaCa-2, Panc-1, and ASPC-1 cells were treated with Ad/g-TRAIL. Transfection and protein expression were determined by using immunoblotting and identification of GFP with fluorescent microscopy and flow cytometry. Cell viability was determined by proliferation assay. Cell-cycle analysis and quantification of caspase-3 were used to identify apoptosis. The in vivo efficacy of Ad/g-TRAIL was characterized in a novel red fluorescent protein murine model of MIA-PaCa-2 pancreatic cancer.

RESULTS:

Cells treated with Ad/g-TRAIL expressed GFP and exhibited apoptotic morphology within 2 days of treatment. Treatment with this vector in vitro resulted in less cell viability, increased caspase-3 activity, and a greater apoptotic fraction than treatment with controls. In vivo, treatment with Ad/g-TRAIL significantly suppressed tumor growth.

CONCLUSIONS:

TRAIL gene therapy induces apoptosis of pancreatic tumor cells both in vitro and in vivo and is a promising therapy in the treatment of pancreatic cancer.

• Authors: Matthew H Katz, Daniel E Spivack, Shinako Takimoto, Bingliang Fang, Douglas W Burton, A R Moossa, Robert M Hoffman, Michael Bouvet
• Journal: Annals of surgical oncology (Ann Surg Oncol) Vol. 10 Issue 7 Pg. 762-72 (Aug 2003) ISSN: 1068-9265 [Print] United States
• PMID: 12900367 (Publication Type: Journal Article)

Chemical References

• Apoptosis Regulatory Proteins
• DNA-Binding Proteins
• Luminescent Proteins
• Membrane Glycoproteins
• TNF-Related Apoptosis-Inducing Ligand
• TNFSF10 protein, human
• Tnfsf10 protein, mouse
• Tumor Necrosis Factor-alpha
• red fluorescent protein
• Green Fluorescent Proteins
• Telomerase
• CASP3 protein, human
• Casp3 protein, mouse
• Caspase 3
• Caspases

Topics

• Adenoviridae
• Animals
• Apoptosis Regulatory Proteins
• Blotting, Western
• Carcinoma, Pancreatic Ductal (therapy)
• Caspase 3
• Caspases (metabolism)
• Catalytic Domain
• DNA-Binding Proteins
• Genes, Transgenic, Suicide
• Genetic Therapy
• Green Fluorescent Proteins
• Humans
• Insect Vectors
• Luminescent Proteins (metabolism, therapeutic use)
• Membrane Glycoproteins (therapeutic use)
• Mice
• Pancreatic Neoplasms (therapy)
• TNF-Related Apoptosis-Inducing Ligand
• Telomerase (therapeutic use)
• Transfection
• Tumor Cells, Cultured
• Tumor Necrosis Factor-alpha (therapeutic use)