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Bid is cleaved upstream of caspase-8 activation during TRAIL-mediated apoptosis in human osteosarcoma cells

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Abstract

TRAIL induces apoptosis in many malignant cell types. In this study, we used the human papilloma virus (HPV) 16 E6 protein as a molecular tool to probe the TRAIL pathway in HCT116 colon carcinoma cells and U2OS osteosarcoma cells. Intriguingly, we found that while E6 protected HCT116 cells from TRAIL, U2OS cells expressing E6 remained sensitive to TRAIL. Furthermore, silencing FADD and procaspase-8 expression with siRNA did not prevent TRAIL-induced apoptosis in U2OS cells. However, siBid provided significant protection from TRAIL, and the cleavage kinetics of Bid and caspase-8 revealed that Bid was cleaved prior to the activation of caspase-8. Cathepsin B activity in U2OS cells was significantly activated shortly after exposure to TRAIL, and the cathepsin B inhibitor, CA074Me, inhibited both TRAIL- and anti-DR5-mediated apoptosis and delayed the cleavage of Bid. These findings suggest that TRAIL activates a pathway dependent on Bid, but largely independent of FADD and caspase-8, in U2OS cells.

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Acknowledgments

This work was supported by NIH Grant 1 R01 CA-095461 (PDH), the Ruth L. Kirschstein National Service Award Individual Fellowship 1 F31 CA113650-01A1 (TG), and by NIH award 2R25 GM060507-05. In addition, we thank Dr. Carl Ware (La Jolla Institute for Allergy and Immunology) for the pcDNA-FADD-encoding plasmid.

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  1. Department of Biochemistry and Microbiology, Loma Linda University School of Medicine, 11085 Campus Street, 121 Mortensen Hall, Loma Linda, CA, 92354, USA

    Theodore O. Garnett, Maria Filippova & Penelope Jayne Duerksen-Hughes

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  1. Theodore O. Garnett
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  2. Maria Filippova
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  3. Penelope Jayne Duerksen-Hughes
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Correspondence to Penelope Jayne Duerksen-Hughes.

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Garnett, T.O., Filippova, M. & Duerksen-Hughes, P.J. Bid is cleaved upstream of caspase-8 activation during TRAIL-mediated apoptosis in human osteosarcoma cells. Apoptosis 12, 1299–1315 (2007). https://doi.org/10.1007/s10495-007-0058-8

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