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在培养的L1210细胞中,用[2-¹⁴C]脱氧胞苷对DNA的胸苷和脱氧胞苷碱基进行标记。

Labelling of the thymidine and deoxycytidine bases of DNA by [2-14C]deoxycytidine in cultured L1210 cells.

作者信息

Karle J M, Hoerauf R M, Cysyk R L

出版信息

Cancer Lett. 1983 Jun;19(2):147-57. doi: 10.1016/0304-3835(83)90148-9.

Abstract

Exposure of cultured L1210 cells to [2-14C]deoxycytidine and analysis or radioactivity incorporated into DNA-pyrimidines revealed that 2.7--5.5-fold more radioactivity is incorporated into DNA-thymine than into cytosine bases. Thus the pathway involving deamination of deoxycytidylate to deoxyuridylate and methylation to thymidylate is highly favored over successive phosphorylation to dCTP. Several modified and endogenous pyrimidines altered the labelling of DNA-thymine and DNA-cytosine with [2-14C]deoxycytidine. 3-Deazauridine at 0.1 mM caused a 56% increase in the labelling of DNA-thymine. Both thymidine and 3-deazauridine (greater than or equal to 10 microM) increased the specific activity of DNA-cytosine by 4-fold. Cytosine arabinoside (ara-C) (greater than or equal to 10 microM) reduced the labelling of both DNA-cytosine and DNA-thymine. Excess cytidine (0.1 mM) reduced the labelling of DNA-cytosine by 40%. Tetrahydrouridine at concentrations up to 1 mM had no effect.

摘要

将培养的L1210细胞暴露于[2-¹⁴C]脱氧胞苷,并分析掺入DNA嘧啶中的放射性,结果显示掺入DNA胸腺嘧啶中的放射性比掺入胞嘧啶碱基中的放射性多2.7至5.5倍。因此,与连续磷酸化生成dCTP相比,涉及脱氧胞苷酸脱氨生成脱氧尿苷酸并甲基化生成胸苷酸的途径受到极大青睐。几种修饰的和内源性嘧啶改变了[2-¹⁴C]脱氧胞苷对DNA胸腺嘧啶和DNA胞嘧啶的标记。0.1 mM的3-去氮尿苷使DNA胸腺嘧啶的标记增加了56%。胸苷和3-去氮尿苷(≥10 μM)均使DNA胞嘧啶的比活性增加了4倍。阿糖胞苷(ara-C)(≥10 μM)降低了DNA胞嘧啶和DNA胸腺嘧啶的标记。过量的胞苷(0.1 mM)使DNA胞嘧啶的标记降低了40%。浓度高达1 mM的四氢尿苷没有影响。

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