Retroviral transfer of deoxycytidine kinase into tumor cell lines enhances nucleoside toxicity.
Deoxycytidine kinase (dCK) phosphorylates a number of nucleoside analogues that are useful in the treatment of various malignancies. Although the level of dCK activity in malignant cells is thought to correlate with chemotherapeutic response, no direct data are available to support this assumption. We have tested this hypothesis by infecting three tumor cell lines, MCF-7, HT-29, and H1437, with the retroviral vector LNPO containing either dCK or LacZ cDNA and measuring the corresponding sensitivity to nucleoside analogues. DCK activity was increased by 1.7-, 2.3-, and 16-fold in MCF-7, HT-29, and H1437 cells, respectively. Northern and Western blots demonstrated a similar increase in mRNA and protein levels. As a result of dCK expression, MCF-7 cells demonstrated a 2.5-fold increase in drug sensitivity to 1-beta-D-arabinofuranosylcytosine (AraC) and 2-chloro-2'-deoxyadenosine (CdA). HT-29 cells had a 7-fold increase in sensitivity to AraC, CdA, and 2-fluoro-9-beta-D-arabinofuranosyladenine, whereas H1437 cells demonstrated a 20- to 106-fold increase. For all three drugs, there was a linear relationship between dCK activity in clonally derived cell lines and IC50s. These data demonstrate a direct effect of dCK activity on drug sensitivity in cell lines. Because many tumors have relatively low levels of dCK, it is possible that dCK gene transfer will be a useful adjunct to the treatment of these malignancies.
Hapke
D M
DM
Department of Pharmacology, University of North Carolina-Chapel Hill, 27599, USA.
Stegmann
A P
AP
Mitchell
B S
BS
D000230:Adenocarcinoma / Q000473:pathology; D000964:Antimetabolites, Antineoplastic / Q000378:metabolism / Q000494:pharmacology*; D001711:Biotransformation / Q000187:drug effects; D001943:Breast Neoplasms / Q000473:pathology; D018288:Carcinoma, Small Cell / Q000473:pathology; D017338:Cladribine / Q000378:metabolism / Q000494:pharmacology*; D003110:Colonic Neoplasms / Q000473:pathology; D003561:Cytarabine / Q000378:metabolism / Q000494:pharmacology*; D018076:DNA, Complementary / Q000235:genetics; D003842:Deoxycytidine Kinase / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism*; D019008:Drug Resistance, Neoplasm; D004354:Drug Screening Assays, Antitumor; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D008175:Lung Neoplasms / Q000473:pathology; D008979:Moloney murine leukemia virus / Q000235:genetics; D010766:Phosphorylation; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000187:drug effects; D014740:Vidarabine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology
D000964:Antimetabolites, Antineoplastic; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D003561:Cytarabine; D017338:Cladribine; D003842:Deoxycytidine Kinase; D014740:Vidarabine; C024352:fludarabine