Caffeine increases sensitivity of DNA to denaturation in chromatin of L1210 cells.
Exposure of exponentially growing L1210 cells to 5 mM and higher concentrations of caffeine perturbs their progression through the cell cycle and results in increased sensitivity of DNA in situ to denaturation. The latter is detected by the increased metachromatic stainability of DNA with acridine orange (AO) and sensitivity to S1 nuclease, measured by flow cytometry. Decreased DNA stability is generally characteristic of chromatin condensation and in untreated cells is observed in mitosis or quiescence (G0). The caffeine-induced decrease in DNA stability affects the interphase cells regardless of their position in the cycle and the changes are stochastic, concentration- and time-dependent. Populations of cells responding to caffeine are very heterogenous with respect to the degree of destabilization of DNA; sensitivity of DNA to denaturation of the maximally affected cells is similar to that of untreated cells in mitosis. The present method allows one to quantitatively express effects of caffeine on nuclear chromatin in individual cells of large cell populations and may be employed in studies correlating chromatin changes induced by this agent with its effects in modulation of cell sensitivity to radiation or antitumour drugs.
Memorial Sloan-Kettering Cancer Center, New York, N.Y. 10021.
J E
JE
Kunicka
A
A
Myc
M R
MR
Melamed
Z
Z
Darzynkiewicz
D000165:Acridine Orange; D000818:Animals; D002110:Caffeine / Q000494:pharmacology*; D002843:Chromatin / Q000187:drug effects*; D004273:DNA, Neoplasm / Q000032:analysis / Q000187:drug effects*; D005434:Flow Cytometry; D007399:Interphase; D007939:Leukemia L1210 / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D009691:Nucleic Acid Denaturation / Q000187:drug effects*; D013194:Staining and Labeling; D014407:Tumor Cells, Cultured
D002843:Chromatin; D004273:DNA, Neoplasm; D002110:Caffeine; D000165:Acridine Orange