On the complexity and dynamics of in vivo Cisplatin-DNA adduct formation using HPLC/ICP-MS.
On the complexity and dynamics of in vivo Cisplatin-DNA adduct formation using HPLC/ICP-MS.
复制标题
DOI：
10.1039/c2mt20128c
复制
发表时间：
2012-10
期刊：
影响因子：
--
通讯作者：
Matthias Ziehe;D. Esteban-Fernández;U. Hochkirch;J. Thomale;M. Linscheid
中科院分区：
文献类型：
--
作者：
Matthias Ziehe;D. Esteban-Fernández;U. Hochkirch;J. Thomale;M. Linscheid
在这项工作中，我们提出了一种方法来测量顺铂与DNA的相互作用所造成的复杂加合物光谱。通过使用优化的DNA消化程序，我们能够表明，在体内双链体DNA中的加合物光谱比迄今为止所描述的要复杂得多。首次使用HPLC/ESI-MS检测到高丰度的链间加合物。这些加合物可能在DNA修复机制和细胞对顺铂耐药性的发展中发挥关键作用。通过物种非特异性同位素稀释分析HPLC/ICP-MS测量，我们能够研究加合物形成的动力学。通过这些实验，我们证明了在加合物初始形成后，DNA链上发生重排，导致加合物模式随时间发生显着变化。此外，优化了物种非特异性同位素稀释分析的参数，以允许测量顺铂暴露细胞DNA中的特异性加合物。
In this work we present a methodology to measure the complex adduct spectrum caused by the interaction of Cisplatin with DNA. By using an optimized DNA digestion procedure we were able to show that the adduct spectrum in in vivo duplex DNA is much more complex than described so far. For the first time a high abundance of interstrand adducts has been detected by using HPLC/ESI-MS. These adducts could play a key role in the DNA repair mechanisms and the development of cellular resistance to Cisplatin. By species-unspecific isotope dilution analysis HPLC/ICP-MS measurements, we were able to study the kinetics of adduct formation. With these experiments we proved that after the initial formation of adducts a rearrangement occurs on the DNA-strands leading to significant changes in adduct patterns over time. Furthermore, the parameters of the species-unspecific isotope dilution analysis were optimized to allow measurements of specific adducts in the DNA of Cisplatin exposed cells.