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Bcl-2-mediated potentiation of neocarzinostatin-induced apoptosis: requirement for caspase-3, sulfhydryl groups, and cleavable Bcl-2

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Abstract

Overexpression of antiapoptotic Bcl-2 family members is thought to contribute to chemotherapeutic resistance of neural crest tumors. Paradoxical potentiation by Bcl-2 of apoptosis induced by the antineoplastic prodrug, neocarzinostatin (NCS), has been observed in PC12 pheochromocytoma cells. Prior studies have indicated that the cleavage of Bcl-2 to its proapoptotic counterpart mediated by caspase-3 is responsible for this potentiation of apoptosis. This has led to the hypothesis that induction of caspase-3 expression in bcl-2-transfected, caspase-3-deficient MCF-7 cells, will result in Bcl-2 cleavage and Bcl-2-dependent potentiation of NCS-induced apoptosis. These studies have further led to the hypothesis that both cleavable Bcl-2 and sulfhydryl groups are required for the activity of caspase-3 in this regard. As hypothesized, co-transfection of bcl-2-transfected MCF-7 cells with a caspase-3 expression construct results in cleavage of Bcl-2 and potentiation of dose-dependent, NCS-mediated cell death. Furthermore, PC12 cells transfected with an expression construct for cleavage-resistant Bcl-2 demonstrated attenuated potentiation of apoptosis relative to their counterparts transfected with wild-type bcl-2. Finally, irreversible oxidative titration of sulfhydryl groups resulted in concentration-dependent attenuation of apoptosis in PC12 cells, along with prevention of caspase-3 activation and Bcl-2 cleavage. These results definitively demonstrate the requirement for caspase-3, cleavable Bcl-2, and available sulfhydryl groups (separate from those required for NCS activation) in potentiation of NCS-induced apoptosis by Bcl-2.

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Acknowledgements

The authors thank Carmel Portugal and Karen D. Nylander for expert technical assistance in the performance of the studies described. They also thank Dr. Guodong Cao for his help in making the caspase-3 expression plasmid. These studies were supported by a grant from the National Institutes of Health (R01-CA74289) and the Carol Ann Craumer Endowment Fund of the Children’s Hospital of Pittsburgh Foundation.

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Authors and Affiliations

  1. Division of Child Neurology, Pediatric Center for Neuroscience, Children’s Hospital of Pittsburgh of UPMC, 3705 Fifth Avenue, Pittsburgh, PA, 15213, USA

    Zhiping Mi, Borah Hong, Zeljka Korade Mirnics, Ye Liang & Nina Felice Schor

  2. Department of Pediatrics, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Zhiping Mi, Borah Hong, Zeljka Korade Mirnics, Ye Liang & Nina Felice Schor

  3. Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Yulia Y. Tyurina & Valerian E. Kagan

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  1. Zhiping Mi
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  2. Borah Hong
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  3. Zeljka Korade Mirnics
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  4. Yulia Y. Tyurina
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  5. Valerian E. Kagan
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  6. Ye Liang
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  7. Nina Felice Schor
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Correspondence to Nina Felice Schor.

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Mi, Z., Hong, B., Mirnics, Z.K. et al. Bcl-2-mediated potentiation of neocarzinostatin-induced apoptosis: requirement for caspase-3, sulfhydryl groups, and cleavable Bcl-2. Cancer Chemother Pharmacol 57, 357–367 (2006). https://doi.org/10.1007/s00280-005-0054-z

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  • DOI: https://doi.org/10.1007/s00280-005-0054-z

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