Percorrer por autor "Pires, Filipe Fernandes"
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- Co-60 source: a study on induction of damages and repair kinetics in a PC3 cell linePublication . Pires, Filipe Fernandes; Belchior, Ana; Eiras, MargaridaThis dissertation was designed to address the study of induction and repair kinetics of Co-60-induced deoxyribonucleic acid (DNA) damages in human prostate tumor cells (PC3 cell line). The analysis aims to investigate the number of complex damage induced in this cell line, to test its repairability and its influence on cell survival capacity. It is intended that this study contributes to the characterization of the response to a specific radiation source by these malignant prostate tumor cells. The present work proposes to evaluate the radio-induced effects in the following 24 hours and to observe the integrity of the survival capacity. In the first part, an immunofluorescence assay is performed. Two biomarkers are used, phosphorylated H2AX (g-H2AX) and tumor suppressor p53 binding protein 1 (53BP1), in order to determine the number of double-strand breaks (DSBs) through their correspondence with the foci, identified by antibodies specific to the biomarkers, over several time-points from the first half hour after irradiation up to 24 hours. Subsequently, co-localization between g-H2AX and 53BP1 is tested using the correlation coefficient provided by CellProfiler. In the second part, a clonogenic assay is performed, observing the evolution of the survival fraction with increasing dose. The results showed that cellular repair after induction of damage allows a decrease in the number of DSBs, but up to 24 hours’ post-irradiation there is a level of residual damage present. Between g-H2AX and 53BP1 there appears to be a partial level of co-localization, with a tendency for decreasing throughout the repair process. In summary, the work described in this dissertation demonstrates the level of induced damage and repair kinetics of the PC3 cell line, suggesting a dose and time repair dependence.