https://scholars.lib.ntu.edu.tw/handle/123456789/370365
標題: | Probing the Dynamics of Doxorubicin-DNA Intercalation during the Initial Activation of Apoptosis by Fluorescence Lifetime Imaging Microscopy (FLIM) | 作者: | Chen, N.-T. Wu, C.-Y. Chung, C.-Y. Hwu, Y. Cheng, S.-H. CHUNG-YUAN MOU Lo, L.-W. |
公開日期: | 2012 | 卷: | 7 | 期: | 9 | 來源出版物: | PLoS ONE | 摘要: | Doxorubicin is a potent anthracycline antibiotic, commonly used to treat a wide range of cancers. Although postulated to intercalate between DNA bases, many of the details of doxorubicin's mechanism of action remain unclear. In this work, we demonstrate the ability of fluorescence lifetime imaging microscopy (FLIM) to dynamically monitor doxorubicin-DNA intercalation during the earliest stages of apoptosis. The fluorescence lifetime of doxorubicin in nuclei is found to decrease rapidly during the first 2 hours following drug administration, suggesting significant changes in the doxorubicin-DNA binding site's microenvironment upon apoptosis initiation. Decreases in doxorubicin fluorescence lifetimes were found to be concurrent with increases in phosphorylation of H2AX (an immediate signal of DNA double-strand breakage), but preceded activation of caspase-3 (a late signature of apoptosis) by more than 150 minutes. Time-dependent doxorubicin FLIM analyses of the effects of pretreating cells with either Cyclopentylidene-[4-(4-chlorophenyl)thiazol-2-yl)-hydrazine (a histone acetyltransferase inhibitor) or Trichostatin A (a histone deacetylase inhibitor) revealed significant correlation of fluorescence lifetime with the stage of chromatin decondensation. Taken together, our findings suggest that monitoring the dynamics of doxorubicin fluorescence lifetimes can provide valuable information during the earliest phases of doxorubicin-induced apoptosis; and implicate that FLIM can serve as a sensitive, high-resolution tool for the elucidation of intercellular mechanisms and kinetics of anti-cancer drugs that bear fluorescent moieties. ? 2012 Chen et al. |
URI: | http://www.scopus.com/inward/record.url?eid=2-s2.0-84866508874&partnerID=MN8TOARS http://scholars.lib.ntu.edu.tw/handle/123456789/370365 |
DOI: | 10.1371/journal.pone.0044947 | SDG/關鍵字: | caspase 3; doxorubicin; genomic DNA; histone H2AX; hydrazine derivative; trichostatin A; apoptosis; article; chromatin condensation; controlled study; double stranded DNA break; drug DNA binding; drug DNA interaction; drug DNA intercalation; enzyme activation; enzyme activity; fluorescence lifetime imaging microscopy; fluorescence microscopy; HeLa cell; histone phosphorylation; in vitro study; Antineoplastic Agents; Apoptosis; Caspase 3; Cell Nucleus; DNA; Doxorubicin; Enzyme Activation; Extracellular Space; HeLa Cells; Humans; Intercalating Agents; Kinetics; Microscopy, Fluorescence; Time Factors |
顯示於: | 化學系 |
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Probing the Dynamics of Doxorubicin-DNA Interc.pdf | 1.07 MB | Adobe PDF | 檢視/開啟 |
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