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.
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.
Katz
Matthew H
MH
Department of Surgery, University of California at San Diego, San Diego, California 92161, USA.
Spivack
Daniel E
DE
Takimoto
Shinako
S
Fang
Bingliang
B
Burton
Douglas W
DW
Moossa
A R
AR
Hoffman
Robert M
RM
Bouvet
Michael
M
D000256:Adenoviridae; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D015153:Blotting, Western; D021441:Carcinoma, Pancreatic Ductal / Q000628:therapy*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D020134:Catalytic Domain; D004268:DNA-Binding Proteins; D041722:Genes, Transgenic, Suicide; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins; D006801:Humans; D007303:Insect Vectors; D008164:Luminescent Proteins / Q000378:metabolism / Q000627:therapeutic use*; D008562:Membrane Glycoproteins / Q000627:therapeutic use*; D051379:Mice; D010190:Pancreatic Neoplasms / Q000628:therapy*; D053221:TNF-Related Apoptosis-Inducing Ligand; D019098:Telomerase / Q000627:therapeutic use*; D014162:Transfection; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000627:therapeutic use*; D000097573:Red Fluorescent Protein
D051017:Apoptosis Regulatory Proteins; D053148:Caspase 3; D020169:Caspases; D004268:DNA-Binding Proteins; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D053221:TNF-Related Apoptosis-Inducing Ligand; D019098:Telomerase; D014409:Tumor Necrosis Factor-alpha; D000097573:Red Fluorescent Protein; C505653:TNFSF10 protein, human; C505654:Tnfsf10 protein, mouse; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse