暴露于乙基化剂后人淋巴细胞和粒细胞加合物和碱不稳定位点的诱导和修复比较

Henk H. Schutte, Govert P. van der Schans, Paul H.M. Lohman
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引用次数: 18

摘要

对暴露于乙基化剂n -乙基-n -亚硝基脲(ENU)和硫酸二乙酯(DES)的人淋巴细胞和粒细胞DNA中加合物和碱不稳定位点的诱导和修复进行了比较研究。为了评估这些损害,用高3h标记的ENU和DES处理人血细胞,并通过HPLC分析所得的3h -乙基加合物。在非放射性ENU和DES处理过程中,DNA中引入的碱不稳定位点通过碱性洗脱检测,并对洗脱组分中的DNA进行荧光定量。所有已知的ENU和DES诱导的加合物均可通过HPLC检测到。此外,由于所用色谱柱的分辨率提高,这些加合物从许多未识别的化合物中分离出来。大多数加合物在处理后20 h的潜伏期内相当持久,但部分加合物(7-乙基腺苷和3-乙基腺苷)部分消失。在淋巴细胞和粒细胞中碱不稳定位点的诱导是非常相似的,但这些位点的去除动力学似乎有很大的不同。在粒细胞中几乎没有任何修复,而在淋巴细胞中,特别是在ENU治疗后,观察到大量且相对较快的修复。在0°C时,人淋巴细胞和粒细胞中碱不稳定位点也发生了诱导;数据表明,这种损伤不是酶修复过程的结果。淋巴细胞和粒细胞对碱不稳定部位的诱导和修复与各种乙基加合物的比较并没有给出可能导致碱不稳定的加合物的身份的线索。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparison of induction and repair of adducts and of alkali-labile sites in human lymphocytes and granulocytes after exposure to ethylating agents

A comparative study has been made of the induction and repair of adducts and alkali-labile sites in the DNA of human lymphocytes and granulocytes exposed to the ethylating agents N-ethyl-N-nitrosourea (ENU) and diethyl sulphate (DES).

To evaluate these damages, the human blood cells were treated with highly 3H-labelled ENU and DES, and the resulting 3H-ethyl adducts were analysed via HPLC. Alkali-labile sites introduced in the DNA during treatment with non-radioactive ENU and DES were detected by alkaline elution with fluorometric quantitation of the DNA in the eluted fractions.

All known adducts induced by ENU and DES could be detected by the HPLC methods applied. Furthermore, these adducts were separated from a number of unidentified compounds, because of the improved resolution on the columns used. Most of the adducts were rather persistent during a subsequent incubation period of up to 20 h after treatment, but some partly disappeared (7-ethyladenine and 3-ethyladenine).

The induction of alkali-labile sites in lymphocytes and granulocytes was very similar, but the kinetics of the removal of these sites appeared to be quite different. In granulocytes there was hardly any repair, whereas in lymphocytes, particularly after ENU treatment, a substantial and relatively fast repair was observed. Induction of alkali-labile sites in human lymphocytes and granulocytes occurred also at 0°C; the data suggest that this kind of damage is not a result of enzymic repair processes.

A comparison of the induction and the repair of alkali-labile sites in lymphocytes and granulocytes with those of the various ethyl adducts did not give a clue as to the identity of the adduct that could be responsible for the lability towards alkali.

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