Wortmannin enhances activation of CPP32 (Caspase-3) induced by TNF or anti-Fas

Fujita, Eriko; Kouroku, Yoriko; Miho, Yasuko; Tsukahara, Toshifumi; Ishiura, Shouichi; Momoi, Takashi

doi:10.1038/sj.cdd.4400355

Cited by 40 publications

(25 citation statements)

References 34 publications

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“…The PI3K inhibitor wortmannin enhanced the activation of caspase-3 that was induced by TNF or anti-Fas (8). Here we demonstrate that TNF acts like other growth factors to promote the phosphorylation of BAD at Ser-136.…”

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confidence: 56%

Tumor Necrosis Factor Induces Phosphorylation and Translocation of BAD through a Phosphatidylinositide-3-OH Kinase-dependent Pathway

Pastorino

Tafani

Farber

1999

Journal of Biological Chemistry

132

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mannin. The mechanism by which the inhibition of the phosphorylation of BAD is likely linked to the induction of lethal mitochondrial damage in TNF-intoxicated cells is discussed. Phosphatidylinositide-3-OH kinase (PI3K)1 is a member of a signaling cascade that eventuates in the phosphorylation of the proapoptotic protein BAD (1, 2). Such growth factors as NGF and interleukin-3 are presumed to promote cell survival as a consequence of the PI3K-mediated activation of the serinethreonine kinase Akt, which in turn phosphorylates BAD (3, 4). Whereas BAD can be phosphorylated at either the Ser-112 or Ser-136 site (5), Akt phosphorylates BAD specifically at Ser-136 (phospho-BAD-136) (2, 6). BAD is capable of forming heterodimers with the antiapoptotic proteins Bcl-X L or Bcl-2 (7). Importantly, phospho-BAD-136 cannot bind either Bcl-X L or Bcl-2 (5). It is postulated that by binding to Bcl-X L and Bcl-2 BAD antagonizes their antiapoptotic activity. Upon inducing its phosphorylation, growth factors lead to the dissociation of BAD from Bcl-X L and Bcl-2 and thereby promote cell survival by allowing the unhindered action of these proteins.The PI3K inhibitor wortmannin enhanced the activation of caspase-3 that was induced by TNF or anti-Fas (8). Here we demonstrate that TNF acts like other growth factors to promote the phosphorylation of BAD at Ser-136. As a result, there is the translocation of BAD from the mitochondria to the cytosol. Moreover, the phosphorylation of BAD by TNF occurs by a PI3K-dependent pathway and is necessary to prevent the cytotoxicity of this cytokine. Inhibition of PI3K prevents both the phosphorylation of BAD and its translocation from mitochondria to the cytosol, effects that are accompanied by substantially enhanced cell killing by TNF. EXPERIMENTAL PROCEDURESTissue Culture-HeLa cells (ATCC-CC-1, American Type Culture Collection) were maintained in 25 cm 2 polystyrene flasks (Corning Costar Corp., Oneonta, NY) with 5 ml of Dulbecco's modified Eagle's medium (DMEM) (high glucose, without pyruvate) (Life Technologies, Inc.) containing 100 units/ml penicillin, 0.1 mg/ml streptomycin, and 10% heat-inactivated fetal bovine serum, all incubated under an atmosphere of 95% air and 5% CO 2 . For transient transfections, HeLa cells were plated at one-third of confluency (3.0 ϫ 10 4 cells/cm 2 ) in 24-well plates. After overnight incubation, the cells were washed twice in phosphate-buffered saline (PBS). Transfections were performed using Lipofectamine-Plus (Life Technologies, Inc.) according to the manufacturer's instructions. Transfection efficiencies of 25-30% were routinely obtained, as assessed by ␤-galactosidase staining. For experiments, cells were transfected with 0.5 g of pCDNA-LacZ and 5 g of either pCDNA, pCDNA3-myc-⌬p85 (referred to as pCDNA-PI3K(Ϫ)

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mentioning

confidence: 56%

Tumor Necrosis Factor Induces Phosphorylation and Translocation of BAD through a Phosphatidylinositide-3-OH Kinase-dependent Pathway

Pastorino

Tafani

Farber

1999

Journal of Biological Chemistry

132

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“…Ac-DEVD-MCA cleavage activity was measured as described previously. 38 After incubation, cells were washed twice with PBS, and the cell pellets were lysed in PBS containing 0.2% Triton X-100 on ice for 10 min. Cells were then centrifuged at 10 0006g for 5 min.…”

Section: Methodsmentioning

confidence: 99%

BMP-4 and retinoic acid synergistically induce activation of caspase-9 and cause apoptosis of P19 embryonal carcinoma cells cultured as a monolayer

et al. 1999

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“…Indeed, PI3-kinase inhibition enhances processing of caspase 3 in TNF-and Fas-treated, but not untreated cells. 95 …”

Section: Pkb and Caspasesmentioning

confidence: 99%

Molecular signals in anti-apoptotic survival pathways

O'Gorman

Cotter

2001

Leukemia

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Drug resistance, to date, has primarily been attributed to increased drug export or detoxification mechanisms. Despite correlations between drug export and drug resistance, it is increasingly apparent that such mechanisms cannot fully account for chemoresistance in neoplasia. It is now widely accepted that chemotherapeutic drugs kill tumour cells by inducing apoptosis, a genetically regulated cell death programme. Evidence is emerging that the exploitation of survival pathways, which may have contributed to disease development in the first instance, may also be important in the development of the chemoresistance. This review discusses the components of and associations between multiple signalling cascades and their possible contribution to the development of neoplasia and the chemoresistant phenotype. Leukemia (2001) 15, 21-34.

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Wortmannin enhances activation of CPP32 (Caspase-3) induced by TNF or anti-Fas

Cited by 40 publications

References 34 publications

Tumor Necrosis Factor Induces Phosphorylation and Translocation of BAD through a Phosphatidylinositide-3-OH Kinase-dependent Pathway

Tumor Necrosis Factor Induces Phosphorylation and Translocation of BAD through a Phosphatidylinositide-3-OH Kinase-dependent Pathway

BMP-4 and retinoic acid synergistically induce activation of caspase-9 and cause apoptosis of P19 embryonal carcinoma cells cultured as a monolayer

Molecular signals in anti-apoptotic survival pathways

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