TY - JOUR
T1 - Combined Cytosine Deaminase Expression, 5-Fluorocytosine Exposure, and Radiotherapy Increases Cytotoxicity to Cholangiocarcinoma Cells
AU - Pederson, Lee C.
AU - Vickers, Selwyn M.
AU - Buchsbaum, Donald J.
AU - Kancharla, Sreekanth R.
AU - Mayo, Matthew S.
AU - Curiel, David T.
AU - Stackhouse, Murray A.
N1 - Funding Information:
From the Departments of Surgery (L.C.P. and SM.V) and Radiation Oncology (D.J.B., S.R.K, and M.A.S.), and the Biostatistics Unit (M.S.M.), and the Gene Therapy Program (D.T.C.), University of Alabama at Birmingham, Birmingham Ala. Supported by award 633602 from the Robert Wood Johnson Foundation. Presented at the Thirty-Eighth Annual Meeting of The Society for Surgery of the Alimentary Tract, Washington, D.C., May 1 l-14,1997. Reprint requests: Selwyn M. Vickers, M.D., University of Alabama at Birmingham, Department of Surgery, Section of Gastrointestinal Surgery, 405 Kracke Building, 1922 Seventh Ave. South, Birmingham, AL 35294-0007. E-mail: [email protected].
PY - 1998
Y1 - 1998
N2 - Cholangiocarcinoma is a malignancy that is resistant to current therapy. We applied the toxin gene therapy strategy of cytosine deaminase conversion of the nontoxic prodrug 5-fluorocytosine to 5-fluorouracil combined with radiotherapy to cholangiocarcinoma. The transduction efficiency of SK-ChA-1 cholangiocarcinoma cells was determined by fluorescence-activated cell-sorting analysis following infection with recombinant adenovirus AdCMVLacZ, which encodes the gene for β-galactosidase. To evaluate cytosine deaminase-mediated conversion of 5-fluorocytosine to 5-fluorouracil and subsequent cytotoxicity, SK-ChA-1 cells were infected with the recombinant adenovirus AdCMVCD, which encodes cytosine deaminase, and exposed to 5-fluorocytosine for 6 to 8 days. Additive cytotoxicity of radiation therapy was evaluated by cobalt-60 exposure following AdCMVCD infection and 5-fluorocytosine treatment. SK-ChA-1 cells were transduced (98.4%) by AdCMVLacZ at 100 plaque-forming units per cell. Following infection with AdCMVCD and exposure to 5 to 100 μg/ml of 5-fluorocytosine, 20% to 64% of SK-ChA-1 cells were killed. A combination of radiation and cytosine deaminase/5-fluorocytosine therapy resulted in enhanced cell killing (83.5% to 91.5%). Cholangiocarcinoma cells were transduced by recombinant adenoviral vectors and were killed by cytosine deaminase-mediated production of 5-fluorouracil. Enhanced cytotoxicity was seen with the addition of external beam radiation. These results provide a foundation for multimodality therapy for human cholangiocarcinoma that combines gene therapy technology with radiation therapy.
AB - Cholangiocarcinoma is a malignancy that is resistant to current therapy. We applied the toxin gene therapy strategy of cytosine deaminase conversion of the nontoxic prodrug 5-fluorocytosine to 5-fluorouracil combined with radiotherapy to cholangiocarcinoma. The transduction efficiency of SK-ChA-1 cholangiocarcinoma cells was determined by fluorescence-activated cell-sorting analysis following infection with recombinant adenovirus AdCMVLacZ, which encodes the gene for β-galactosidase. To evaluate cytosine deaminase-mediated conversion of 5-fluorocytosine to 5-fluorouracil and subsequent cytotoxicity, SK-ChA-1 cells were infected with the recombinant adenovirus AdCMVCD, which encodes cytosine deaminase, and exposed to 5-fluorocytosine for 6 to 8 days. Additive cytotoxicity of radiation therapy was evaluated by cobalt-60 exposure following AdCMVCD infection and 5-fluorocytosine treatment. SK-ChA-1 cells were transduced (98.4%) by AdCMVLacZ at 100 plaque-forming units per cell. Following infection with AdCMVCD and exposure to 5 to 100 μg/ml of 5-fluorocytosine, 20% to 64% of SK-ChA-1 cells were killed. A combination of radiation and cytosine deaminase/5-fluorocytosine therapy resulted in enhanced cell killing (83.5% to 91.5%). Cholangiocarcinoma cells were transduced by recombinant adenoviral vectors and were killed by cytosine deaminase-mediated production of 5-fluorouracil. Enhanced cytotoxicity was seen with the addition of external beam radiation. These results provide a foundation for multimodality therapy for human cholangiocarcinoma that combines gene therapy technology with radiation therapy.
UR - http://www.scopus.com/inward/record.url?scp=0032057198&partnerID=8YFLogxK
U2 - 10.1016/S1091-255X(98)80024-3
DO - 10.1016/S1091-255X(98)80024-3
M3 - Article
C2 - 9841986
AN - SCOPUS:0032057198
SN - 1091-255X
VL - 2
SP - 283
EP - 291
JO - Journal of Gastrointestinal Surgery
JF - Journal of Gastrointestinal Surgery
IS - 3
ER -