Abstract

Methods of fluorescence spectroscopy and microscopy—including intensity and lifetime (FLIM) images—are used to examine uptake, intracellular location and interaction of the chemotherapeutic drug doxorubicin in MCF-7 human breast cancer cells as a function of cholesterol content. By comparing cells with natural and decreased cholesterol levels after 2 h or 24 h incubation with doxorubicin, we observed that higher fluorescence intensities and possibly shortened fluorescence lifetimes—reflecting increased uptake of the drug and more pronounced drug response—are concomitant with higher membrane fluidity.

Highlights

  • IntroductionDoxorubicin, an anthracycline antibiotic, is used as a cytostatic drug in cancer chemotherapy, such as breast cancer, bronchial carcinoma and lymphoma, and has been studied for several decades [1,2]

  • Doxorubicin, an anthracycline antibiotic, is used as a cytostatic drug in cancer chemotherapy, such as breast cancer, bronchial carcinoma and lymphoma, and has been studied for several decades [1,2].The drug is taken up by cells due to passive diffusion through their membrane and intercalates in DNA strands, where it causes chromatin condensation and initiates apoptosis [3]

  • Intensity of Doxorubicin Fluorescence Increases after Cholesterol Depletion

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Summary

Introduction

Doxorubicin, an anthracycline antibiotic, is used as a cytostatic drug in cancer chemotherapy, such as breast cancer, bronchial carcinoma and lymphoma, and has been studied for several decades [1,2]. The drug is taken up by cells due to passive diffusion through their membrane and intercalates in DNA strands, where it causes chromatin condensation and initiates apoptosis [3]. Due to its fluorescence properties [4] doxorubicin can be localized within the cells, e.g., by wide-field microscopy, and, fluorescence lifetime measurements [5,6,7,8] permit assessing. 2013, 14 intermolecular interactions with its microenvironment. Low or moderate light doses are needed to avoid phototoxic effects in microscopic experiments [9]

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