Chapter 14 Measurement of Cell Death

Loo, Deryk; Rillema, Jill R.

doi:10.1016/s0091-679x(08)61583-6

Cited by 184 publications

(129 citation statements)

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“…In the report by Ashkenazi et al, 1999, PrEC were found to be TRAIL-insensitive, judging by propidium iodide staining. A possible drawback to the use of vital dyes, such as propidium iodide, is that cells undergoing apoptosis may retain cell membrane integrity and appear alive until late in the apoptotic program when secondary necrosis begins (Loo and Rillema, 1998). For example, in our experiments, staining of PrEC with vital dye, trypan blue, did not detect a signi®cant number of dead cells if they were treated with TRAIL for less than 24 h (data not shown).…”

mentioning

confidence: 60%

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) triggers apoptosis in normal prostate epithelial cells

Nesterov

Ivashchenko²,

Kraft

2002

Oncogene

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mentioning

confidence: 60%

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) triggers apoptosis in normal prostate epithelial cells

Nesterov

Ivashchenko²,

Kraft

2002

Oncogene

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“…DNA Fragmentation-Apoptosis was accompanied by fragmentation of DNA (25)(26). To determine the occurrence of DNA fragmentation, total DNA was extracted from unstimulated and stimulated KS cells.…”

Section: Methodsmentioning

confidence: 99%

HIV-1-Tat Protein Activates Phosphatidylinositol 3-Kinase/ AKT-dependent Survival Pathways in Kaposi's Sarcoma Cells

Deregibus¹,

Cantaluppi²,

Doublier³

et al. 2002

Journal of Biological Chemistry

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In this study we found that Tat protected vincristinetreated Kaposi's sarcoma cells from apoptosis and from down-regulation of several anti-apoptotic genes such as AKT-1, AKT-2, BCL2, BCL-XL, and insulin-like growth factor I and induced the de novo expression of the interleukin-3 gene. Moreover, we found that Tat enhanced phosphorylation of AKT and BAD proteins. The inhibition of phosphatidylinositol 3-kinase with two unrelated pharmacological inhibitors, wortmannin and LY294002, abrogated both the anti-apoptotic effect and the phosphorylation of AKT induced by Tat. After treatment with Tat, the AKT enzymatic activity showed a biphasic increase: an early activation (15 min), independent from protein synthesis; and a delayed activation (24 h), which was significantly decreased upon blockage of protein synthesis. Experiments with a function blocking antivascular endothelial cell growth factor receptor-2 antibody suggested that both the early and delayed AKT activation and the protection from apoptosis were triggered by the interaction of Tat with vascular endothelial cell growth factor receptor-2. Moreover, experiments with function-blocking antibodies directed against insulin-like growth factor I/insulin-like growth factor I receptor or interleukin-3 indicated their involvement in the delayed activation of AKT and their contribution to the anti-apoptotic effect of Tat on vincristine-treated Kaposi's sarcoma cells.

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“…7) A ladder pattern of internucleosomal DNA fragmentation was observed when 10 nmol/l of AETD was applied to the cells (Fig. 1B).…”

Section: )mentioning

confidence: 93%

“…The following traditional criteria for this purpose have been proposed: (1) typical morphologic changes, including chromatin condensation and nuclear fragmentation, must be observed under light microscopy; (2) a DNA ladder must be demonstrated by agarose gel electrophoresis. 7) Several new assays potentially useful for the detection of apoptosis have recently been developed on the basis of biochemical events associated with apoptosis (e.g. DNA fragmentation by a flow cytometry, 6) terminal transferase-mediated dUTP nick endlabeling (TUNEL) staining, 7) and caspase activation 8) ).…”

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confidence: 99%

“…7) Several new assays potentially useful for the detection of apoptosis have recently been developed on the basis of biochemical events associated with apoptosis (e.g. DNA fragmentation by a flow cytometry, 6) terminal transferase-mediated dUTP nick endlabeling (TUNEL) staining, 7) and caspase activation 8) ). Several apoptosis-inducing agents isolated from the natural products have been reported to have anti-cancer potential.…”

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confidence: 99%

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4-Acetyl-12,13-epoxyl-9-trichothecene-3,15-diol Isolated from the Fruiting Bodies of Isaria japonica YASUDA Induces Apoptosis of Human Leukemia Cells (HL-60).

Hong

et al. 2001

Biological & Pharmaceutical Bulletin

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The fruiting bodies of Isaria fungi have been traditionally used in Korea to treat cancer. An apoptosis-inducing compound, 4-acetyl-12,13-epoxyl-9-trichothecene-3,15-diol, was isolated from the methanol extract of fruiting bodies of Isaria japonica YASUDA by bioassay-guided fractionation. The apoptosis of the human leukemia cells (HL-60) by the compound was accessed by propidium iodide-staining flow cytometric analysis, and apoptosis-inducing activity at IC 50 concentration (10 nmol/l) was further confirmed by a nuclear morphological change, a ladder pattern of internucleosomal DNA fragmentation, and an activation of caspase-3.Key words Isaria japonica YASUDA; human HL-60 leukemia cell; 4-acetyl-12,13-epoxyl-9-trichothecene-3,15-diol; apoptosis japonica were inoculated into potato dextrose agar, and incubated at 25°C until the fungal colony grow to 5 cm in diameter. It usually took 10 d, and the colony turned gray. The cultures were then refrigerated at approximately 4°C, and used as stock cultures. In the second culture, a 1 l flask, containing 500 ml of culture medium, which had been sterilized at 121°C for 20 min and then cooled to 15°C, was inoculated with four 5-mm agar plugs taken from stock cultures. The flask culture was aerated by agitation on an orbital shaker at 140 rpm for a period of 3 d at 25°C, then at 160 ppm for 4 d. The culture medium consists of potato (200 g/l) and sugars (30 g/l). All cultures were conducted in the dark room. The third fermentation was carried out in the pupae of the silkworm (100 g). The fermentation media were autoclaved at 121°C for 40 min. After the media had cooled to 15°C, they were inoculated with 5 ml of spore inoculum and incubated for 8 d at 25°C in the dark room. After 8 d of cultivation, the culture condition was changed to 60% humidity for 10 d in the dark room, then 80% humidity at 20°C in 2000 lux of brightness for 30 d. After incubation, the fruiting bodies of the fungi were harvested, and extracted with MeOH for further chemical and biological investigations.Isolation and Structure Determination of 4-Acetyl-12,13-epoxy-9-trichothecene-3,15-diol The air-dried fruiting bodies (226 g) from the fermentation of I. japonica were extracted with methanol (MeOH) for 48 h. The MeOH extract was concentrated, suspended in H 2 O, and sequentially partitioned with n-hexane, CH 2 Cl 2 , and ethyl acetate (EtOAc). The EtOAc-soluble fraction (247.5 mg) was subjected to C18 flash column chromatography with a stepwise gradient of 30 to 100% (v/v) MeOH in H 2 O. The fractions eluting at 30% and 40% MeOH in H 2 O (36.8 mg) were combined, and subjected to preparative reversed-phase HPLC using a gradient from 15 to 35% CH 3 CN in H 2 O over 40 min, then 100% CH 3 CN for 10 min (Alltech HS Hyperprep 100 BDS C 18 (1.0ϫ25 cm; 8-mm particle size; 2 ml/min; UV detection at 210 nm)) to yield compound 1 (12.3 mg). The structure of compound 1 was identified as 4-acetyl-12,13-epoxy-9-trichothecene-3,15-diol (AEDT) by the comparison of MS and NMR data with those reported in the literature ...

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Chapter 14 Measurement of Cell Death

Cited by 184 publications

References 19 publications

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) triggers apoptosis in normal prostate epithelial cells

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) triggers apoptosis in normal prostate epithelial cells

HIV-1-Tat Protein Activates Phosphatidylinositol 3-Kinase/ AKT-dependent Survival Pathways in Kaposi's Sarcoma Cells

4-Acetyl-12,13-epoxyl-9-trichothecene-3,15-diol Isolated from the Fruiting Bodies of Isaria japonica YASUDA Induces Apoptosis of Human Leukemia Cells (HL-60).

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