7-Hydroxicoumarin Exposure Induces Prompt Activation of Caspase-3 in A-549 Cancer Cells Through Single Cell Microinjection
Current Overview on Pharmaceutical Science Vol 7(2023)
摘要
Coumarins and their apoptogenic effects have arisen conspicuous interest in recent years. The present chapter aims to determine whether 7-hydroxycoumarin (7-HC) induces changes in caspase-3 (C-3) activity in A549 human lung carcinoma cells. The effects of 7-HC were investigated using a variety of analytical approaches, including colorimetric and fluorometric assays, western blotting, single-cell microinjection, fluorescence microscopy, and image analysis. After being exposed to 1.85 mM 7-HC for 24 hours, C3 activity increased by 65%, procaspase-3 was significantly converted to C-3, and poly(ADP-ribose) polymerase was cleaved. Additionally, morphological modifications linked to apoptosis were noticed. The calcium conductance of the cells increased by 27% after being exposed to 7HC for 3 or 6 hours. Single-cell microinjection of a specific fluorescent substrate of C-3 into cells that had previously been exposed to 7-HC allowed for the identification of a typical enzymatic kinetic profile of C-3 activation several hours before the morphological and biochemical changes connected to the observation of apoptosis. These findings imply that 7-HC, the most important biotransformation product of coumarin in people, is what causes the rapid in vivo activation of C-3.
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