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COMBINATION OF ATOMIC FORCE MICROSCOPY AND COMET ASSAY FOR ANALYSIS OF DNA DAMAGE INDUCED BY PDT


The aim of the present study was to evaluate the efficiency of photosensitisation induced by two photosensitizers, TMPyP and ClAlPcS2, tested in vitro on the tumor cell line MCF7. The oxidative damage of DNA in MCF-7 cells was analyzed by comet assay (CA) combined with Atomic Force Microscopy (AFM). The ability of detection of apoptotic response detected by Atomic Force Microscopy at the individual molecule level of DNA was successfully demonstrated; when DNA get damaged, cleavage to fragments caused by photodynamic treatment was directly visualized by AFM imaging of individual molecules. Its accuracy and reliability was validated through the comparison with traditional single cell agarose electrophoresis.

Keywords:
photodynamic therapy, atomic force microscopy, DNA damage, comet assay


Autoři: Hana Zapletalová 1;  Kateřina Bartoň Tománková 1;  Jakub Malohlava 1;  Milan Vůjtek 2;  Hana Kolářová 1
Působiště autorů: Department of Medical Biophysics, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Republic 1;  Department of Experimental Physics, Faculty of Science, Palacky University Olomouc, Czech Republic 2
Vyšlo v časopise: Lékař a technika - Clinician and Technology No. 4, 2016, 46, 83-90
Kategorie: Původní práce

Souhrn

The aim of the present study was to evaluate the efficiency of photosensitisation induced by two photosensitizers, TMPyP and ClAlPcS2, tested in vitro on the tumor cell line MCF7. The oxidative damage of DNA in MCF-7 cells was analyzed by comet assay (CA) combined with Atomic Force Microscopy (AFM). The ability of detection of apoptotic response detected by Atomic Force Microscopy at the individual molecule level of DNA was successfully demonstrated; when DNA get damaged, cleavage to fragments caused by photodynamic treatment was directly visualized by AFM imaging of individual molecules. Its accuracy and reliability was validated through the comparison with traditional single cell agarose electrophoresis.

Keywords:
photodynamic therapy, atomic force microscopy, DNA damage, comet assay


Zdroje

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