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  • Original Paper
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Bcl-XL protects pancreatic adenocarcinoma cells against CD95- and TRAIL-receptor-mediated apoptosis

Abstract

In this study we sought to clarify the role of the pro-apoptotic potential of mitochondria in the death pathway emanating from the TRAIL (APO-2L) and CD95 receptors in pancreatic carcinoma cells. We focused on the role of the Bcl-2 family member Bcl-XL, using three pancreatic carcinoma cell lines as a model system, two of which have high (Panc-1, PancTuI) and one has low (Colo357) Bcl-XL expression. In these cell lines, the expression of Bcl-XL correlated with sensitivity to apoptosis induced by TRAIL or anti-CD95. Flow cytometric analysis revealed cell surface expression of TRAIL-R1 and TRAIL-R2 on PancTuI and Colo357, and TRAIL-R2 on Panc-1 cells. In Colo357 cells retrovirally transduced with Bcl-XL, caspase-8 activation in response to treatment with TRAIL or anti-CD95 antibody was not different from parental cells and EGFP-transfected controls, however, apoptosis was completely suppressed as measured by the mitochondrial transmembrane potential Δψm, caspase-3 activity (PARP cleavage) and DNA-fragmentation. Inhibition of Bcl-XL function by overexpression of Bax or administration of antisense oligonucleotides against Bcl-XL mRNA resulted in sensitization of Panc-1 cells to TRAIL and PancTuI cells to anti-CD95 antibody-induced cell death. The results show that Bcl-XL can protect pancreatic cancer cells from CD95- and TRAIL-mediated apoptosis. Thus, in these epithelial tumour cells the mitochondrially mediated ‘type II’ pathway of apoptosis induction is not only operative regarding the CD95 system but also regarding the TRAIL system.

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Abbreviations

ATCC:

American type culture collection

DISC:

death-inducing signalling complex

EGFP:

enhanced green fluorescent protein

FACS:

fluorescence activated cell sorting

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

mAB:

monoclonal (m) antibody (ab)

PAGE:

polyacrylamide gel electrophoresis

PARP:

poly(ADP-ribose) polymerase

TRAIL:

tumor necrosis factor-related apoptosis-inducing ligand

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Acknowledgements

We thank Dr JC Reed for the Bax expression vector, Dr T Ketterer for synthesis of Bcl-X antisense-134 phosphothioate oligonucleotides and Immunex Corp. (Seattle, WA, USA) for the kind gift of anti-TRAIL receptor antibodies. We also thank C Timm, A Schulze, H Stahl, E Rieser for excellent technical assistance, and Dr B Howard, San Diego, for establishing the retroviral transduction techniques. Supported by the Deutsche Forschungsgemeinschaft, SFB 415, project A3 and by Grant KV 9821 and a BioFuture Grant from the Bundesministerium für Bildung und Forschung (BMBF) to H Kalthoff and H Walczak, respectively. Some of the results from this study form part of the doctoral thesis of S Hinz.

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Hinz, S., Trauzold, A., Boenicke, L. et al. Bcl-XL protects pancreatic adenocarcinoma cells against CD95- and TRAIL-receptor-mediated apoptosis. Oncogene 19, 5477–5486 (2000). https://doi.org/10.1038/sj.onc.1203936

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