DNA-Platinum Thin Films for Use in Chemoradiation Therapy Studies.
DNA-Platinum Thin Films for Use in Chemoradiation Therapy Studies.
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DOI：
10.1155/2012/923914
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发表时间：
2012
影响因子：
3.8
通讯作者：
Sanche L
中科院分区：
文献类型：
--
作者：
Rezaee M;Alizadeh E;Hunting D;Sanche L
铂化疗药物与质粒DNA（Pt-DNA）共价结合的干膜是一种有用的实验模型，可用于研究放射对紧邻铂化疗药物的DNA的直接影响，这种情况与了解伴随放化疗的机制具有相当大的相关性。在本文中，我们确定了用于辐照实验的Pt-DNA膜的最佳制备条件。DNA铂化反应的孵育条件对膜中Pt-DNA的结构有实质性的影响。通过电感耦合等离子体质谱法测量作为孵育时间和温度的函数的与DNA结合的Pt的量。我们的实验表明，当DNA铂化在37°C下发生24小时时，Pt-DNA样品中的化学不稳定性和DNA损伤增加，该条件已广泛用于体外研究。用于形成Pt-DNA膜的DNA的铂化在室温下反应时间小于2小时是最佳的。通过增加Pt化合物相对于DNA的浓度，从而加速它们相互结合的速率，可以制备含有已知浓度的Pt的Pt-DNA样品，同时减少由更长的程序引起的DNA降解。
Dry films of platinum chemotherapeutic drugs covalently bound to plasmid DNA (Pt-DNA) represent a useful experimental model to investigate direct effects of radiation on DNA in close proximity to platinum chemotherapeutic agents, a situation of considerable relevance to understand the mechanisms underlying concomitant chemoradiation therapy. In the present paper we determine the optimum conditions for preparation of Pt-DNA films for use in irradiation experiments. Incubation conditions for DNA platination reactions have a substantial effect on the structure of Pt-DNA in the films. The quantity of Pt bound to DNA as a function of incubation time and temperature is measured by inductively coupled plasma mass spectroscopy. Our experiments indicate that chemical instability and damage to DNA in Pt-DNA samples increase when DNA platination occurs at 37°C for 24 hours, the condition which has been extensively used for in vitro studies. Platination of DNA for the formation of Pt-DNA films is optimal at room temperature for reaction times less than 2 hours. By increasing the concentration of Pt compounds relative to DNA and thus accelerating the rate of their mutual binding, it is possible to prepare Pt-DNA samples containing known concentrations of Pt while reducing DNA degradation caused by more lengthy procedures.
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