Metabolism of 5-fluorocytosine to 5-fluorouracil in human colorectal tumor cells transduced with the cytosine deaminase gene: significant antitumor effects when only a small percentage of tumor cells express cytosine deaminase.
The gene encoding cytosine deaminase (CD) has been expressed in the human colorectal carcinoma cell line WiDr. Metabolism studies confirm that tumor cells expressing CD convert the very nontoxic prodrug 5-fluorocytosine (5FCyt) to 5-fluorouracil (5FUra) and 5FUra metabolites. Tumor xenografts composed of CD-expressing cells can selectively generate tumor levels of > 400 microM 5FUra when the host mouse is dosed with nontoxic levels of 5FCyt. The selective metabolic conversion of 5FCyt to 5FUra in CD-expressing tumor cells results in the inhibition of thymidylate synthase and incorporation of 5FUra into RNA. 5FUra is also liberated into the surrounding environment when CD-expressing tumor cells are treated with 5FCyt. The liberated 5FUra is able to kill neighboring, non-CD-expressing tumor cells in vitro and in vivo. Most importantly, when only 2% of the tumor mass contains CD-expressing cells (98% non-CD-expressing cells), significant regressions in all tumors are observed when the host mouse is dosed with nontoxic levels of 5FCyt.
Huber
B E
BE
Division of Cell Biology, Wellcome Research Laboratories, Research Triangle Park, NC 27709.
Austin
E A
EA
Richards
C A
CA
Davis
S T
ST
Good
S S
SS
D000818:Animals; D001711:Biotransformation; D002455:Cell Division; D002470:Cell Survival; D002851:Chromatography, High Pressure Liquid; D015179:Colorectal Neoplasms / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology / Q000628:therapy; D043525:Cytosine Deaminase; D004273:DNA, Neoplasm / Q000096:biosynthesis; D005437:Flucytosine / Q000378:metabolism*; D005472:Fluorouracil / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D051379:Mice; D008819:Mice, Nude; D009700:Nucleoside Deaminases / Q000096:biosynthesis / Q000378:metabolism*; D012334:RNA, Neoplasm / Q000096:biosynthesis; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors; D013997:Time Factors; D014183:Transplantation, Heterologous; D014316:Tritium; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D012334:RNA, Neoplasm; D014316:Tritium; D005437:Flucytosine; D013940:Thymidylate Synthase; D009700:Nucleoside Deaminases; D043525:Cytosine Deaminase; D005472:Fluorouracil