PURPOSE: The purpose of this study was to investigate the effect of calyculin A on the number of γ-H2AX foci (phosphorylated histone variant 2AX) in lymphocytes after in vitro and in vivo irradiation with rather low doses as they are used in diagnostic and interventional radiology. MATERIALS AND METHODS: For in vitro experiments blood samples of 14 healthy volunteers were irradiated with different doses (10, 50, 100 mGy) and incubated with (0.01, 0.1, 1, 10 nM) or without calyculin A for up to 2 hours. Non-irradiated samples with and without calyculin A served as controls. For in vivo evaluation blood samples were collected from seven patients undergoing computed tomography (CT) both with 1 nM calyculin A containing vials and vials without calyculin A. Foci were quantified in isolated lymphocytes using γ-H2AX immunofluorescence microscopy. RESULTS: 1 nM calyculin A led to a complete inhibition of γ-H2AX foci loss in irradiated samples whereas no inhibition of p53 binding protein 1 (53 BP1) foci was found. Lower concentrations of the phosphatase inhibitor did not have a sufficient effect on foci decrease. Calyculin A did not affect foci levels in non-irradiated samples. If no calyculin A was added into the vial before the blood draws detectable CT-induced foci levels were lower in all patients with a reduction of the medians of 35%. CONCLUSIONS: Using γ-H2AX immunofluorescence microscopy calyculin A can be a useful tool to mark the induced γ-H2AX foci after low dose irradiation and to avoid an underestimation of the real deoxyribonucleic acid (DNA) damage in in vitro and in vivo experiments.
PURPOSE: The purpose of this study was to investigate the effect of calyculin A on the number of γ-H2AX foci (phosphorylated histone variant 2AX) in lymphocytes after in vitro and in vivo irradiation with rather low doses as they are used in diagnostic and interventional radiology. MATERIALS AND METHODS: For in vitro experiments blood samples of 14 healthy volunteers were irradiated with different doses (10, 50, 100 mGy) and incubated with (0.01, 0.1, 1, 10 nM) or without calyculin A for up to 2 hours. Non-irradiated samples with and without calyculin A served as controls. For in vivo evaluation blood samples were collected from seven patients undergoing computed tomography (CT) both with 1 nM calyculin A containing vials and vials without calyculin A. Foci were quantified in isolated lymphocytes using γ-H2AX immunofluorescence microscopy. RESULTS: 1 nM calyculin A led to a complete inhibition of γ-H2AX foci loss in irradiated samples whereas no inhibition of p53 binding protein 1 (53 BP1) foci was found. Lower concentrations of the phosphatase inhibitor did not have a sufficient effect on foci decrease. Calyculin A did not affect foci levels in non-irradiated samples. If no calyculin A was added into the vial before the blood draws detectable CT-induced foci levels were lower in all patients with a reduction of the medians of 35%. CONCLUSIONS: Using γ-H2AX immunofluorescence microscopy calyculin A can be a useful tool to mark the induced γ-H2AX foci after low dose irradiation and to avoid an underestimation of the real deoxyribonucleic acid (DNA) damage in in vitro and in vivo experiments.
Authors: Niels Belmans; Anne Caroline Oenning; Benjamin Salmon; Bjorn Baselet; Kevin Tabury; Stéphane Lucas; Ivo Lambrichts; Marjan Moreels; Reinhilde Jacobs; Sarah Baatout Journal: Dentomaxillofac Radiol Date: 2021-05-14 Impact factor: 3.525
Authors: Lucián Zastko; Anna Račková; Petra Petrovičová; Matúš Durdík; Jakub Míšek; Eva Marková; Igor Belyaev Journal: Int J Mol Sci Date: 2021-05-22 Impact factor: 5.923
Authors: Bregje Van Oorschot; Arlene L Oei; Anna C Nuijens; Hans Rodermond; Ron Hoeben; Jan Stap; Lukas J Stalpers; Nicolaas A P Franken Journal: Cell Mol Biol Lett Date: 2013-12-23 Impact factor: 5.787