Caspase-8 activation is necessary but not sufficient for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis in the prostatic carcinoma cell line LNCaP

Oskar W Rokhlin; Natalya V Guseva; Agshin F Tagiyev; Rebecca A Glover; Michael B Cohen

doi:10.1002/pros.10074

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Caspase-8 activation is necessary but not sufficient for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis in the prostatic carcinoma cell line LNCaP

Journal article

Peer reviewed

Caspase-8 activation is necessary but not sufficient for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis in the prostatic carcinoma cell line LNCaP

Oskar W Rokhlin, Natalya V Guseva, Agshin F Tagiyev, Rebecca A Glover and Michael B Cohen

The Prostate, Vol.52(1), pp.1-11

06/01/2002

DOI: 10.1002/pros.10074

PMID: 11992615

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Abstract

The differential sensitivity of tumor cells to TRAIL-induced apoptosis may be mediated by different intracellular inhibitors of apoptosis, and only a few reports have described the pathway(s) that are activated in response to TRAIL in prostate cells. LNCaP was transfected with a dominant-negative form of FADD (FADD-DN) and cells were selected in the presence of hygromycin. Cell viability was estimated by calcein assay. Apoptosis was estimated by caspase activation using both fluorogenic substrates and Western blot analysis of activated caspases. To detect cytochrome c release, mitochondria-free cytosol was prepared and Western blot analysis was performed. LNCaP is resistant to TRAIL but TRAIL transiently induces DEVDase activity and activation of caspase-8; caspase-2, -3, -7, and -9 were not activated. Wortmannin, an inhibitor of the PI3K/Akt pathway, converted the phenotype of LNCaP from TRAIL-resistant to -sensitive. In the presence of wortmannin TRAIL induced activation of caspase-2, -3, -7, -8, and -9, as well as dissipation of mitochondrial transmembrane potential and release of cyto-chrome c from mitochondria into the cytosol. In addition, combined TRAIL and wortmannin treatment resulted in cleavage of several proteins: PARP, Akt, p21/WAF1, and MDM2 as well as dephosphorylation of Akt. The proteolysis of p21/WAFI and Akt, which are known survival factors, presumably amplify the apoptotic cascade in LNCaP. Transfection of FADD-DN in LNCaP resulted in inhibition of caspase activation as well as in resistance to combined treatment with TRAIL and wortmannin. These results suggest that caspase-8 activation is necessary but not sufficient for TRAIL-mediated apoptosis and is presumably blocked downstream of caspase-8 by the PI3K/Akt pathway.

Adaptor Proteins, Signal Transducing

Androstadienes - pharmacology

Apoptosis

Apoptosis Regulatory Proteins

Blotting, Western

Carrier Proteins - genetics

Carrier Proteins - physiology

Caspase 8

Caspase 9

Caspases - metabolism

Cinnamates

Cyclin-Dependent Kinase Inhibitor p21

Cyclins - metabolism

Drug Resistance

Enzyme Activation

Enzyme Inhibitors - pharmacology

Fas-Associated Death Domain Protein

Humans

Hygromycin B - analogs & derivatives

Hygromycin B - pharmacology

Male

Membrane Glycoproteins - pharmacology

Membrane Glycoproteins - physiology

Nuclear Proteins

Peptide Hydrolases - metabolism

Phosphatidylinositol 3-Kinases - antagonists & inhibitors

Phosphorylation

Prostatic Neoplasms - metabolism

Prostatic Neoplasms - pathology

Protein-Serine-Threonine Kinases

Proteins - metabolism

Proto-Oncogene Proteins - metabolism

Proto-Oncogene Proteins c-akt

Proto-Oncogene Proteins c-mdm2

TNF-Related Apoptosis-Inducing Ligand

Transfection

Tumor Cells, Cultured

Tumor Necrosis Factor-alpha - pharmacology

Tumor Necrosis Factor-alpha - physiology

Details

Title: Subtitle: Caspase-8 activation is necessary but not sufficient for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis in the prostatic carcinoma cell line LNCaP
Creators: Oskar W Rokhlin - University of Iowa
Natalya V Guseva - University of Iowa
Agshin F Tagiyev - University of Iowa
Rebecca A Glover - University of Iowa
Michael B Cohen - University of Iowa
Resource Type: Journal article
Publication Details: The Prostate, Vol.52(1), pp.1-11
DOI: 10.1002/pros.10074
PMID: 11992615
ISSN: 0270-4137
eISSN: 1097-0045
Grant note: CA 87717 / NCI NIH HHS
Language: English
Date published: 06/01/2002
Academic Unit: Pathology; Biology
Record Identifier: 9984646762502771

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