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CdSe-ZnS量子点对DNA的光敏断裂与损伤

Photosensitized breakage and damage of DNA by CdSe-ZnS quantum dots.

作者信息

Anas AbdulAziz, Akita Hidetaka, Harashima Hideyoshi, Itoh Tamitake, Ishikawa Mitsuru, Biju Vasudevanpillai

机构信息

Nanobioanalysis Team, Health Technology Research Center, National Institute of Advanced Industrial Science and Technology, 2217-14 Hayashi-cho, Takamatsu, Kagawa 761-0395, Japan.

出版信息

J Phys Chem B. 2008 Aug 14;112(32):10005-11. doi: 10.1021/jp8018606. Epub 2008 Jun 26.

Abstract

Strand breakages and nucleobase damages in plasmid DNA (pDNA) by CdSe-ZnS quantum dots (QDs) are investigated under different conditions of photoactivation. Here, streptavidin functionalized CdSe-ZnS QDs are conjugated to biotinylated pDNA, and photosensitized strand breakages and nucleobase damages in the conjugates are investigated using atomic force microscopy (AFM) imaging, gel electrophoreses analyses, and assay of reactive oxygen intermediates (ROI). Also, reactions of photoactivated pDNA-QD conjugates with base excision repair enzymes such as formamidopyrimidine glycosylase (Fpg) and endonuclease III (Endo III) show damages of purine and pyrimidine bases. The base excision repair enzymes recognize and remove the damaged bases. The base excision reactions of photoactivated pDNA-QD conjugates resulted in pDNA strand breakages, which appeared as sheared bands in agarose gel images. On the basis of AFM imaging, reactions of Fpg and Endo III with damaged pDNA, ROI assay, and literature reports, we attribute the breakage and damage of pDNA to its reactions with ROI. The production of ROI by photoactivated QDs is confirmed by nitroblue tetrazolium (NBT) assay. The current work shows that photoactivation of QD-conjugated nucleic acids for an extended period of time is not favorable for their stability. On the other hand, photoinduced production of ROI by QDs is an emerging research area with potential applications in the photodynamic therapy of cancer. In this regard, photosensitized damage of pDNA observed in the current work shows possibilities of QDs in nucleus-targeted photodynamic therapy.

摘要

在不同的光激活条件下,研究了CdSe-ZnS量子点(QDs)对质粒DNA(pDNA)的链断裂和核碱基损伤。在此,将链霉亲和素功能化的CdSe-ZnS QDs与生物素化的pDNA偶联,并使用原子力显微镜(AFM)成像、凝胶电泳分析和活性氧中间体(ROI)测定来研究偶联物中的光敏链断裂和核碱基损伤。此外,光激活的pDNA-QD偶联物与碱基切除修复酶如甲酰胺嘧啶糖基化酶(Fpg)和内切酶III(Endo III)的反应显示了嘌呤和嘧啶碱基的损伤。碱基切除修复酶识别并去除受损碱基。光激活的pDNA-QD偶联物的碱基切除反应导致pDNA链断裂,这在琼脂糖凝胶图像中表现为剪切带。基于AFM成像、Fpg和Endo III与受损pDNA的反应、ROI测定以及文献报道,我们将pDNA的断裂和损伤归因于其与ROI的反应。通过硝基蓝四唑(NBT)测定证实了光激活的QDs产生ROI。目前的工作表明,长时间光激活QD偶联的核酸对其稳定性不利。另一方面,QDs光诱导产生ROI是一个新兴的研究领域,在癌症光动力治疗中具有潜在应用。在这方面,当前工作中观察到的pDNA的光敏损伤显示了QDs在细胞核靶向光动力治疗中的可能性。

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