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Single-cell microinjection assay indicates that 7-hydroxycoumarin induces rapid activation of caspase-3 in A549 cancer cells

  • Authors:
    • Maribel Soto‑Nuñez
    • Karen Azucena Díaz‑Morales
    • Patricia Cuautle‑Rodríguez
    • Víctor Torres‑Flores
    • José Sullivan López‑González
    • Juan José Mandoki‑Weitzner
    • Juan Arcadio Molina‑Guarneros
  • View Affiliations / Copyright

    Affiliations: Department of Pharmacology, School of Medicine, National University of Mexico, Mexico City 04510, Mexico, Research Division, School of Medicine, National University of Mexico, Mexico City 04510, Mexico, Department of Chronic‑Degenerative Diseases, National Institute of Respiratory Diseases ‘Ismael Cosio Villegas’, Mexican Ministry of Health, Mexico City 14080, Mexico
    Copyright: © Soto‑Nuñez et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 1789-1795
    |
    Published online on: September 23, 2015
       https://doi.org/10.3892/etm.2015.2765
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Abstract

Coumarins have attracted intense interest in recent years due to their apoptogenic effects. The aim of the present study was to determine whether 7-hydroxycoumarin (7-HC) induces changes in caspase-3 (C‑3) activity in A549 human lung carcinoma cells. A range of analytical techniques, including colorimetric and fluorometric assays, western blotting, single‑cell microinjection, fluorescence microscopy and image analysis were conducted to elucidate the effects of 7‑HC. A 24‑h exposure to 1.85 mM 7‑HC induced a 65% increase in C‑3 activity, and a notable conversion of procaspase‑3 to C‑3, in addition to poly(ADP‑ribose)polymerase cleavage. Furthermore, morphological changes associated with apoptosis were observed. Exposure of the cells to 7‑HC for 3 or 6 h increased calcium conductance by 27%. By performing the single‑cell microinjection of a specific fluorescent substrate of C‑3 into previously 7‑HC‑exposed cells, a typical enzymatic kinetic profile of C‑3 activation was identified a number of hours prior to the morphological and biochemical changes associated with apoptosis being observed. These results suggest that the rapid in vivo activation of C‑3 is induced by 7‑HC, the most relevant biotransformation product of coumarin in humans.
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Copy and paste a formatted citation
Spandidos Publications style
Soto‑Nuñez M, Díaz‑Morales KA, Cuautle‑Rodríguez P, Torres‑Flores V, López‑González JS, Mandoki‑Weitzner JJ and Molina‑Guarneros JA: Single-cell microinjection assay indicates that 7-hydroxycoumarin induces rapid activation of caspase-3 in A549 cancer cells. Exp Ther Med 10: 1789-1795, 2015.
APA
Soto‑Nuñez, M., Díaz‑Morales, K.A., Cuautle‑Rodríguez, P., Torres‑Flores, V., López‑González, J.S., Mandoki‑Weitzner, J.J., & Molina‑Guarneros, J.A. (2015). Single-cell microinjection assay indicates that 7-hydroxycoumarin induces rapid activation of caspase-3 in A549 cancer cells. Experimental and Therapeutic Medicine, 10, 1789-1795. https://doi.org/10.3892/etm.2015.2765
MLA
Soto‑Nuñez, M., Díaz‑Morales, K. A., Cuautle‑Rodríguez, P., Torres‑Flores, V., López‑González, J. S., Mandoki‑Weitzner, J. J., Molina‑Guarneros, J. A."Single-cell microinjection assay indicates that 7-hydroxycoumarin induces rapid activation of caspase-3 in A549 cancer cells". Experimental and Therapeutic Medicine 10.5 (2015): 1789-1795.
Chicago
Soto‑Nuñez, M., Díaz‑Morales, K. A., Cuautle‑Rodríguez, P., Torres‑Flores, V., López‑González, J. S., Mandoki‑Weitzner, J. J., Molina‑Guarneros, J. A."Single-cell microinjection assay indicates that 7-hydroxycoumarin induces rapid activation of caspase-3 in A549 cancer cells". Experimental and Therapeutic Medicine 10, no. 5 (2015): 1789-1795. https://doi.org/10.3892/etm.2015.2765
Copy and paste a formatted citation
x
Spandidos Publications style
Soto‑Nuñez M, Díaz‑Morales KA, Cuautle‑Rodríguez P, Torres‑Flores V, López‑González JS, Mandoki‑Weitzner JJ and Molina‑Guarneros JA: Single-cell microinjection assay indicates that 7-hydroxycoumarin induces rapid activation of caspase-3 in A549 cancer cells. Exp Ther Med 10: 1789-1795, 2015.
APA
Soto‑Nuñez, M., Díaz‑Morales, K.A., Cuautle‑Rodríguez, P., Torres‑Flores, V., López‑González, J.S., Mandoki‑Weitzner, J.J., & Molina‑Guarneros, J.A. (2015). Single-cell microinjection assay indicates that 7-hydroxycoumarin induces rapid activation of caspase-3 in A549 cancer cells. Experimental and Therapeutic Medicine, 10, 1789-1795. https://doi.org/10.3892/etm.2015.2765
MLA
Soto‑Nuñez, M., Díaz‑Morales, K. A., Cuautle‑Rodríguez, P., Torres‑Flores, V., López‑González, J. S., Mandoki‑Weitzner, J. J., Molina‑Guarneros, J. A."Single-cell microinjection assay indicates that 7-hydroxycoumarin induces rapid activation of caspase-3 in A549 cancer cells". Experimental and Therapeutic Medicine 10.5 (2015): 1789-1795.
Chicago
Soto‑Nuñez, M., Díaz‑Morales, K. A., Cuautle‑Rodríguez, P., Torres‑Flores, V., López‑González, J. S., Mandoki‑Weitzner, J. J., Molina‑Guarneros, J. A."Single-cell microinjection assay indicates that 7-hydroxycoumarin induces rapid activation of caspase-3 in A549 cancer cells". Experimental and Therapeutic Medicine 10, no. 5 (2015): 1789-1795. https://doi.org/10.3892/etm.2015.2765
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