抗癌药物阿霉素的荧光发射光谱分析

ANALYSIS OF FLUORESCENCE EMISSION SPECTRA OF THE ANTICANCER DRUG DOXORUBICIN

利用蒽环类抗癌药物阿霉素的自发荧光可反映药物在细胞中的分布和相对剂量；利用特异性荧光探针Hoechst33342(Ho．33342）可标记活细胞DNA，从而反映细胞所处周期时相及染色体的形态变化．但是在阿霉素和Ho．33342共存在的细胞中，用MPVⅡ型显微荧光分光光度计对药物的自发荧光和Ho．3342的发射荧光进行光谱分析的结果表明：在紫外激发下，药物自发荧光与Ho.33342荧光之间存在干扰；选择适合的外加带通滤片S525后，这种干扰可降低到可以忽略的程度，从而在同一细胞内可分离开2种荧光，并进行定量分析．该结论有利于今后发展影像分析和荧光多参数相关测量方法，可以通过荧光图像的获取及分析来研究单个活细胞摄取、滞留抗癌药物与细胞增殖、细胞凋亡及抗药性的关系，从而揭示药物的作用机理．

The distribution and relative dose of the anthracycline anticancer drug Doxorubicin in the cell are reflected by its autofluorescence. The DNA of the living cell is marked with the special fluorescent probe Hoechst33342, by which the phase of the cell cycle could be identified and the morphological changes of the chromatin can be observed. However, in the same cell simultaneously marked by Doxorubicin and Ho.33342, the fluorescent emission spectranalysis by MPVⅡ microspectrofluorometer demonstrates that excited by UV, there is interference between the fluorescence of Doxorubicin and Ho.33342. When the proper additional band-pass filter S525 is adopted, the interference could be decreased to a negligible degree, thus, in the same cell, the two kind of fluorescence could be divided and be quantitatively measured. This conclusion devotes to developing the methods of image analysis and fluorescent multiparameters measurement. Through the acquisition and analysis of the fluorescent image, the relation between the uptake, retention of antitumor agent and cell proliferation, cell apoptosis, multidrug resistance (MDR) in single living cell can be investigated so as to discover the drug function mechanism.