Transfer of the cytidine deaminase cDNA into hematopoietic cells.
In order to investigate whether transfer of the cytidine deaminase (CDD) cDNA would increase chemotherapy resistance to cytosine arabinoside (ara-C) we used a retroviral vector expressing both, neomycin phosphotransferase and the CDD cDNA, to transduce hematopoietic cells from cell lines and from murine bone marrow (BM). After coculture on producer clones with a viral titer of 1 x 10(5) CFU/ml and up to 3-fold increased CDD enzymatic activity, WEHI-3 cell line and primary hematopoietic cells were exposed to ara-C in clonogenic assays. A transduction efficiency of 34.8 +/- 6.2% could be determined for BM clonogenic progenitor cells by G418 resistance. We could observe significantly more colonies (77 +/- 3.1%) surviving from transduced primary BM cells than from mock cells (51.7 +/- 9.3%) at 10(-8) mol/l ara-C. At 10(-7) mol/l ara-C 8.7% of BM cells became absolutely resistant after retroviral transduction. Our data confirm that CDD represents another candidate gene for increasing resistance to cytotoxic drugs in hematopoietic cells.
Flasshove
M
M
Department of Internal Medicine (Cancer Research), West German Cancer Center, University of Essen Medical School.
Frings
W
W
Schröder
J K
JK
Moritz
T
T
Schütte
J
J
Seeber
S
S
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D003561:Cytarabine / Q000494:pharmacology; D003564:Cytidine Deaminase / Q000235:genetics*; D017931:DNA Primers; D018076:DNA, Complementary / Q000008:administration & dosage*; D004279:DNA, Viral; D004351:Drug Resistance / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000201:enzymology*; D051379:Mice; D011401:Promoter Regions, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic
D017931:DNA Primers; D018076:DNA, Complementary; D004279:DNA, Viral; D003561:Cytarabine; D003564:Cytidine Deaminase