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Quinacrine induces cytochrome c-dependent apoptotic signaling in human cervical carcinoma cells

  • Research Articles
  • Pharmacology, Toxicology & Pharmaceutic
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Abstract

Quinacrine (QU), a phospholipase-A2 (PLA-2) inhibitor has been used clinically as a chemotherapeutic adjuvant. To understand the mechanisms leading to its chemotherapeutic effect, we have investigated QU-induced apoptotic signaling pathways in human cervical squamous carcinoma Hela cells. In this study, we found that QU induced cytochrome c-dependent apoptotic signaling. The release of pro-apoptotic cytochrome c was QU concentration-and time-dependent, and preceded activation of caspase-9 and −3. Flow cytometric FACScan analysis using fluorescence intensities of DiOC6 demonstrated that QU-induced cytochrome c release was independent of mitochondrial permeability transition (MPT), since the concentrations of QU that induced cytochrome c release did not alter mitochondrial membrane potential (▴ Ψm). Moreover, kinetic analysis of caspase activities showed that cytochrome c release led to the activation of caspase-9 and downstream death effector, caspase-3. Caspase-3 inhibitor (Ac-DEVD-CHO) partially blocked QU-induced apoptosis, suggesting the importance of caspase-3 in this apoptotic signaling mechanism. Supplementation with arachidonic acid (AA) sustained caspase-3 activation induced by QU. Using inhibitors against cellular arachidonate metabolism of lipooxygenase (Nordihydroxyguaiaretic Acid, NDGA) and cyclooxygenase (5, 8, 11,14-Eicosatetraynoic Acid, ETYA) demonstrated that QU-induced apoptotic signaling may be dependent on its role as a PLA-2 inhibitor. Interestingly, NDGA attenuated QU-induced cytochrome c release, caspase activity as well as apoptotic cell death. The blockade of cytochrome c release by NDGA was much more effective than that attained with cyclosporin A (CsA), a MPT inhibitor. ETYA was not effective in blocking cytochrome c release, except under very high concentrations. Caspase inhibitor z-VAD blocked the release of cytochrome c suggesting that this signaling event is caspase dependent, and caspase-8 activation may be upstream of the mitochondrial, events. In summary, we report that QU induced cytochrome c-dependent apoptotic signaling cascade, which may be dependent on its role as a PLA-2 inhibitor. This apoptotic mechanism induced by QU may contribute to its known chemotherapeutic effects.

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Author notes

  1. Adedigbo A. Fasanmade

    Present address: Centocor Biotechnologies Inc., 200 Great Valley Parkway, 19355, Malvern, PA, USA

Authors and Affiliations

  1. Department of Pharmaceutics and Pharmacodynamics, Center for Pharmaceutical Biotechnology, College of Pharmacy, 200 Great Valley Parkway, 19355, Malvern, PA, USA

    Adedigbo A. Fasanmade, Edward D. Owuor, Rachel P. L. Ee, Dima Qato & Ah-Ng Tony Kong

  2. Centocor Biotechnologies Inc., 200 Great Valley Parkway, 19355, Malvern, PA, USA

    Mark Heller

  3. Department of Pharmaceutics, College of Pharmacy, Rutgers University, Environmental and Occupational Health Sciences Institute, 170 Frelinghuysen Road, 08854, Piscataway, NJ, USA

    Edward D. Owuor & Ah-Ng Tony Kong

  4. Department of Hematology/Oncology, College of Medicine, University of Illinois at Chicago, 60607, Chicago, IL, USA

    Adedigbo A. Fasanmade, Edward D. Owuor, Rachel P. L. Ee, Dima Qato, Mark Heller & Ah-Ng Tony Kong

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  1. Adedigbo A. Fasanmade
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Correspondence to Ah-Ng Tony Kong.

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Fasanmade, A.A., Owuor, E.D., Ee, R.P.L. et al. Quinacrine induces cytochrome c-dependent apoptotic signaling in human cervical carcinoma cells. Arch. Pharm. Res. 24, 126–135 (2001). https://doi.org/10.1007/BF02976480

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  • DOI: https://doi.org/10.1007/BF02976480

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