Characterization of L5178Y murine lymphoblasts resistant to quinone antitumor agents.
The exact contribution of the quinone group to the activity of quinone antitumor agents remains uncertain. Two L5178Y murine lymphoblastic cell lines resistant to the model quinone antitumor agent, hydrolyzed benzoquinone mustard, and one partial-revertant cell line were isolated and characterized. The antitumor activity of hydrolyzed benzoquinone mustard has been shown previously to be due to its ability to induce free radical mediated DNA strand breaks. Resistant cells were obtained by growing a cloned L5178Y parental cell line in media containing increasing concentrations of hydrolyzed benzoquinone mustard. L5178Y/HBM2 cells were selected from L5178Y cells growing in media containing 0.2 mM drug, while L5178Y/HBM10 cells were selected from cells growing in media containing 1.0 mM drug. The L5178Y/HBMR cells were obtained by growing L5178Y/HBM10 cells in media without hydrolyzed benzoquinone mustard. The resistant cell lines, L5178Y/HBM2 and L5178Y/HBM10, were 2.5- and 6-fold less sensitive, respectively, to hydrolyzed benzoquinone mustard compared to parental cells, and this was accompanied by a decrease in the formation of DNA single and double strand breaks by this drug. The partial-revertant cell line, L5178Y/HBMR was 2.9-fold less sensitive to hydrolyzed benzoquinone mustard compared to parental cells. Drug uptake appeared to be lower in the resistant cells compared to parental cells. The resistant cells had a slightly elevated level of superoxide dismutase activity compared to parental cells, but there was no increase in the mRNA for superoxide dismutase nor any amplification of the gene for this enzyme. Intracellular catalase activities of the L5178Y/HBM2 and L5178Y/HBM10 cells were elevated by 1.25- and 2.6-fold, respectively, and the increased enzyme activity in the L5178Y/HBM10 cells appeared to result from a 3.6-fold increase in mRNA for this enzyme. Glutathione peroxidase activity was slightly elevated in L5178Y/HBM2 cells, but was unchanged in the other resistant cells. The L5178Y/HBM2 and L5178Y/HBM10 cells showed increased concentrations of glutathione and elevated levels of glutathione transferase activity. The resistant cell lines also had DT-diaphorase activity that was 3- and 24-fold higher in L5178Y/HBM2 and L5178Y/HBM10 cells, respectively, compared to sensitive cells. However, cytochrome P-450 reductase activity and the ratio of reduced to oxidized pyridine nucleotides was unchanged in the resistant cell lines. The partial-revertant cell line, L5178Y/HBMR, showed approximately the same level of resistance to hydrolyzed benzoquinone mustard as the L5178Y/HBM2 cells.(ABSTRACT TRUNCATED AT 400 WORDS)
D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology*; D016227:Benzoquinones; D001692:Biological Transport; D002470:Cell Survival / Q000187:drug effects; D003579:Cytochrome Reductases / Q000378:metabolism; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000187:drug effects; D004351:Drug Resistance; D005979:Glutathione Peroxidase / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D007940:Leukemia L5178 / Q000503:physiopathology*; D007942:Leukemia, Experimental / Q000503:physiopathology*; D009243:NAD / Q000378:metabolism; D016660:NAD(P)H Dehydrogenase (Quinone); D009249:NADP / Q000378:metabolism; D011808:Quinone Reductases / Q000378:metabolism; D011809:Quinones / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism
D000970:Antineoplastic Agents; D016227:Benzoquinones; D004273:DNA, Neoplasm; D011809:Quinones; D013438:Sulfhydryl Compounds; D009243:NAD; C040043:2-(bis(2-hydroxyethyl)amino)-1,4-benzoquinone; D009249:NADP; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; D003579:Cytochrome Reductases; D016660:NAD(P)H Dehydrogenase (Quinone); D011808:Quinone Reductases; D005982:Glutathione Transferase