Mesh : Animals Cell Line Chromatography / methods Chromatography, Ion Exchange Cricetinae Cricetulus Culture Media DNA / metabolism Thymidine / analogs & derivatives metabolism

来  源:   DOI:10.1093/carcin/1.6.487

Abstract:
When Chinese hamster V79A cells are cultured in the presence of O4-methyl-[6-3H]-thymidine the incorporation of this modified nucleoside into newly synthesised DNA is observed. The radioactivity incorporated has been identified as O4-methylthymidine by digesting the DNA to 3\'-monophosphates with spleen phosphodiesterase followed by treatment with alkaline phosphatase to give O4-methylthymidine as the major radioactive product. The radioactivity associated with the latter co-chromatographs with authentic O4-methylthymidine in several chromatographic systems. Once incorporated the modified nucleoside appears to be rapidly removed from the DNA with half-life of 2-3 h. There is no evidence of demethylation of the O4-methylthymidine to give thymidine, in either the culture medium or within the cells once incorporated into DNA. Although the levels of incorporation observed are low, (being only 125 O4-methylthymidine residues per 10(8) thymidine residues at a modified nucleoside concentration of 10(-5) M), they may still be relevant as similar levels are apparently produced in the DNA of the cultured cells on treatment with biologically significant doses of carcinogenic alkylating agents.
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