4-hydroxynonenal induces a cellular redox status-related activation of the caspase cascade for apoptotic cell death

2000 ◽  
Vol 113 (4) ◽  
pp. 635-641 ◽  
Author(s):  
W. Liu ◽  
M. Kato ◽  
A.A. Akhand ◽  
A. Hayakawa ◽  
H. Suzuki ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Death ◽  
Dna Fragmentation ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
K562 Cells ◽  
Apoptotic Cell Death ◽  
Jurkat Cells ◽  
Substrate Peptide ◽  
Caspase Cascade

4-Hydroxynonenal (HNE), a diffusible product of lipid peroxidation, has been suggested to be a key mediator of oxidative stress-induced cell death. In this study, we partially characterized the mechanism of HNE-mediated cytotoxicity. Incubation of human T lymphoma Jurkat cells with 20–50 μM HNE led to cell death accompanied by DNA fragmentation. Western blot analysis showed that HNE-treatment induced time- and dose-dependent activation of caspase-8, caspase-9 and caspase-3. HNE-induced caspase-3 processing was confirmed by a flow cytometric demonstration of increased catalytic activity on the substrate peptide. HNE treatment also led to remarkable cleavage of poly(ADP-ribose) polymerase (PARP), which was prevented by pretreatment of cells with DEVD-FMK as a caspase-3 inhibitor. The HNE-mediated activation of caspases, cleavage of PARP and DNA fragmentation were blocked by antioxidants cysteine, N-acety-L-cysteine and dithiothreitol, but not by two other HNE-reactive amino acids lysine and histidine, or by cystine, the oxidized form of cysteine. HNE rapidly decreased levels of intracellular reduced glutathione (GSH) and its oxidized form GSSG, and these were also attenuated by the reductants. Coincubation of Jurkat cells with a blocking anti-Fas antibody prevented Fas-induced but not HNE-induced activation of caspase-3. HNE also activated caspase-3 in K562 cells that do not express functional Fas. Our results thereby demonstrate that HNE triggers oxidative stress-linked apoptotic cell death through activation of the caspase cascade. The results also suggest a possible mechanism involving a direct scavenge of intracellular GSH by HNE.

The AKT Inhibitor, A443654, Induces Cell Cycle Arrest, Apoptosis and Synergizes with Chemotherapeutic Drugs in Multi-Drug Resistant T-Cell Acute Lymphoblastic Leukemia - A Novel Agent for Therapy of Drug Resistant ALL.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3344-3344
Author(s):  
James A. McCubrey ◽  
William L. Blalock ◽  
Pier Luigi Tazzari ◽  
Alessandra Cappellini ◽  
Ilaria Iacobucci ◽  
...  
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Apoptotic Cell Death ◽  
Jurkat Cells ◽  
Drug Resistant ◽  
Akt Inhibitor ◽  
Cycle Arrest ◽  
Hematopoietic Precursor ◽  
Dose Dependent

Abstract The PI3K/Akt pathway is a key mediator of proliferation and survival; its constitutive activation is implicated in pathogenesis and progression of a wide variety of hematological malignancies. Constitutively-activated Akt is a common feature of the T-ALL cell lines (MOLT-4, Jurkat, CEM). Hyperactivity of Akt leads not only to their growth and resistance to apoptosis but also to drug-resistance. In this study, we sought to examine the potential of a novel Akt inhibitor, A443654, in the therapy of T-ALL. A443654 led to rapid cell death of CEM, Jurkat, and MOLT-4. All three lines were sensitive to nanomolar (nM) doses of A443654 (IC50=60 nM, 120 nM and 900 nM for MOLT-4, CEM, and Jurkat, respectively). Effects were dose dependent and resulted in G2/M arrest. Indeed, approximately 39% of Jurkat cells treated with A443654 were in G2/M whereas only 11% of the untreated cells were in G2/M. Cell cycle arrest was followed by apoptotic cell death as determined by annexin V-PI and trypan blue staining. Treatment of CEM and Jurkat cells led to de-phosphorylation of the downstream Akt substrate GSK-3beta. Treatment of Jurkat cells with A443654 resulted in activation of caspase-2, -3, -8, and -9. Apoptotic cell death was greatly reduced by caspase-3 and -9 selective inhibitors. Additionally, A443654 was shown to be highly effective against the drug-resistant cell line CEM-VBL100 (CEM-R), which overexpresses Pgp. This Akt inhibitor initially (1 to 4 hour treatments) decreased Pgp activity, but not protein levels, further documenting a link between Akt and Pgp activity. While CEM-R cells displayed decreased sensitivity to A443654, treatment of CEM-R cells with sub-lethal doses of A443654 for 24 hours, reduced the surface expression of Pgp. Moreover, A443654 synergized with the DNA damaging agent etoposide, a substrate of Ppg, in both drug sensitive and resistant lines. At etoposide concentrations between 25 mM to 100 mM, A443654 enhanced the extent of cell killing from 25% to 45%. We then confirmed the effect of A443654 on human leukemia samples using blasts from 6 patients with T-ALL, all of whom displayed constitutive Akt activation. In a dose dependent fashion, A443654 was able to induce apoptotic cell death of T-ALL blast cells, as indicated by flow cytometric analysis of samples immunostained for active (cleaved) caspase-3. In contrast, this Akt inhibitor was determined to be minimally cytotoxic on normal CD34+ hematopoietic precursor cells isolated from cord blood. Taken together, our findings indicate that the Akt inhibitor, A443654, either alone or in combination with existing drugs, may in the future be a useful therapeutic option for primary and refractory T-ALL displaying activated Akt signaling. Furthermore, this novel Akt inhibitor was effective in suppressing the growth of multidrug resistant ALL cells while having minimal effects on normal hematopoietic precursor cells documenting its poteintial in the treatment of drug resistant leukemias.

Berberine Induces Apoptotic Cell Death via Activation of Caspase-3 and -8 in HL-60 Human Leukemia Cells: Nuclear Localization and Structure–Activity Relationships

2017 ◽  
Vol 45 (07) ◽  
pp. 1497-1511 ◽  
Author(s):  
Shinya Okubo ◽  
Takuhiro Uto ◽  
Aya Goto ◽  
Hiroyuki Tanaka ◽  
Tsuyoshi Nishioku ◽  
...  
Keyword(s):  
Cell Death ◽  
Dna Fragmentation ◽  
Antiproliferative Activity ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Apoptotic Cell Death ◽  
Leukemia Cells ◽  
Caspase 8 ◽  
Human Leukemia ◽  
Caspase 9

Berberine (BBR), an isoquinoline alkaloid, is a well-known bioactive compound contained in medicinal plants used in traditional and folk medicines. In this study, we investigated the subcellular localization and the apoptotic mechanisms of BBR were elucidated. First, we confirmed the incorporation of BBR into the cell visually. BBR showed antiproliferative activity and promptly localized to the nucleus from 5[Formula: see text]min to 15[Formula: see text]min after BBR treatment in HL-60 human promyelocytic leukemia cells. Next, we examined the antiproliferative activity of BBR (1) and its biosynthetically related compounds (2-7) in HL-60 cells. BBR exerted strongest antiproliferative activity among 1-7 and the results of structures and activity relation suggested that a methylenedioxyl group in ring A, an [Formula: see text]-alkyl group at C-9 position, and the frame of isoquinoline may be necessary for antiproliferative activity. Moreover, BBR showed the most potent antiproliferative activity in HL-60 cells among human cancer and normal cell lines tested. Next, we examined the effect of BBR on molecular events known as apoptosis induction. In HL-60 cells, BBR induced chromatin condensation and DNA fragmentation, and triggered the activation of PARP, caspase-3 and caspase-8 without the activation of caspase-9. BBR-induced DNA fragmentation was abolished by pretreatment with inhibitors against caspase-3 and caspase-8, but not against caspase-9. ERK and p38 were promptly phosphorylated after 15 min of BBR treatment, and this was correlated with time of localization to the nucleus of BBR. These results demonstrated that BBR translocated into nucleus immediately after treatments and induced apoptotic cell death by activation of caspase-3 and caspase-8.

Comparison of endothelial function, O2−· and H2O2 production, and vascular oxidative stress resistance between the longest-living rodent, the naked mole rat, and mice

2006 ◽  
Vol 291 (6) ◽  
pp. H2698-H2704 ◽  
Author(s):  
Nazar Labinskyy ◽  
Anna Csiszar ◽  
Zsuzsanna Orosz ◽  
Kira Smith ◽  
Aracelie Rivera ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cell Death ◽  
Endothelial Function ◽  
Dna Fragmentation ◽  
Apoptotic Cell ◽  
Heat Exposure ◽  
Apoptotic Cell Death ◽  
Vascular Aging ◽  
Mole Rat ◽  
Naked Mole Rat

Vascular aging is characterized by decreased nitric oxide (NO) bioavailability, oxidative stress, and enhanced apoptotic cell death. We hypothesized that interspecies comparative assesment of vascular function among rodents with disparate longevity may offer insight into the mechanisms determining successful vascular aging. We focused on four rodents that show approximately an order of magnitude range in maximum longevity (ML). The naked mole rat (NMR; Heterocephalus glaber) is the longest-living rodent known (ML > 28 yr), Damara mole rats (DMRs, Cryptomys damarensis; ML ∼ 16 yr) and guinea pigs (GPs, Cavia porcellus; ML ∼ 6 yr) have intermediate longevity, whereas laboratory mice are short living (ML ∼ 3.5 yr). We compared interspecies differences in endothelial function, O2−· and H2O2 production, and resistance to apoptotic stimuli in blood vessels. Sensitivity to acetylcholine-induced, NO-mediated relaxation was smaller in carotid arteries from NMRs, GPs, and DMRs than in mouse vessels. Measurements of production of O2−· (lucigenin chemiluminescence and ethidium bromide fluorescence) and H2O2 (dichlorofluorescein fluorescence) showed that free radical production in vascular endothelial and smooth muscle cells is comparable in vessels of the three longer-living species and in arteries of shorter-living mice. In mouse arteries, H2O2 (from 10−6 to 10−3 mol/l) and heat exposure (42°C for 15–45 min) enhanced apoptotic cell death, as indicated by an increased DNA fragmentation rate and increased caspase 3/7 activity. In NMR vessels, only the highest doses of H2O2 enhanced apoptotic cell death, whereas heat exposure did not increase DNA fragmentation rate. Interspecies comparison showed there is a negative correlation between H2O2-induced apoptotic cell death and ML. Thus endothelial vasodilator function and vascular production of reactive oxygen species do not correlate with maximal lifespan, whereas increased lifespan potential is associated with an increased vascular resistance to proapoptotic stimuli.

scholarly journals Active caspase 3 and DNA fragmentation as markers for apoptotic cell death in primary and metastatic liver tumours

Pathology ◽  
2007 ◽  
Vol 39 (6) ◽  
pp. 558-564 ◽  
Author(s):  
Eva Karamitopoulou ◽  
Luca Cioccari ◽  
Sabine Jakob ◽  
Claudio Vallan ◽  
Thomas Schaffner ◽  
...  
Keyword(s):  
Cell Death ◽  
Dna Fragmentation ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Apoptotic Cell Death ◽  
Liver Tumours ◽  
Metastatic Liver ◽  
Active Caspase

scholarly journals Transcriptome Analysis Reveals HgCl2 Induces Apoptotic Cell Death in Human Lung Carcinoma H1299 Cells through Caspase-3-Independent Pathway

2021 ◽  
Vol 22 (4) ◽  
pp. 2006
Author(s):  
Mi Jin Kim ◽  
Jinhong Park ◽  
Jinho Kim ◽  
Ji-Young Kim ◽  
Mi-Jin An ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Transcriptome Analysis ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Expression Patterns ◽  
Nitrogen Compound ◽  
Apoptotic Cell Death ◽  
Mercury Chloride ◽  
Rna Seq

Mercury is one of the detrimental toxicants that can be found in the environment and exists naturally in different forms; inorganic and organic. Human exposure to inorganic mercury, such as mercury chloride, occurs through air pollution, absorption of food or water, and personal care products. This study aimed to investigate the effect of HgCl2 on cell viability, cell cycle, apoptotic pathway, and alters of the transcriptome profiles in human non-small cell lung cancer cells, H1299. Our data show that HgCl2 treatment causes inhibition of cell growth via cell cycle arrest at G0/G1- and S-phase. In addition, HgCl2 induces apoptotic cell death through the caspase-3-independent pathway. Comprehensive transcriptome analysis using RNA-seq indicated that cellular nitrogen compound metabolic process, cellular metabolism, and translation for biological processes-related gene sets were significantly up- and downregulated by HgCl2 treatment. Interestingly, comparative gene expression patterns by RNA-seq indicated that mitochondrial ribosomal proteins were markedly altered by low-dose of HgCl2 treatment. Altogether, these data show that HgCl2 induces apoptotic cell death through the dysfunction of mitochondria.

Two steroidal saponins from Camassia cusickii induce L1210 cell death through the apoptotic mechanism

2001 ◽  
Vol 79 (11) ◽  
pp. 953-958 ◽  
Author(s):  
Ellyawati Candra ◽  
Kimihiro Matsunaga ◽  
Hironori Fujiwara ◽  
Yoshihiro Mimaki ◽  
Yutaka Sashida ◽  
...  
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Chromatin Condensation ◽  
Flow Cytometric Analysis ◽  
Apoptotic Cell Death ◽  
Morphological Observation ◽  
Steroidal Saponin ◽  
Steroidal Saponins ◽  
L1210 Cells

Two steroidal saponins, tigogenin hexasaccharide-1 (TGHS-1, (25R)-5α-spirostan-3β-yl 4-O-[2-O-[3-O- (α-L-rhamnopyranosyl)-β-D-glucopyranosyl]-3-O-[4-O-(α-L-rhamnopyranosyl)-β-D-glucopyranosyl]-β-D-glucopyranosyl]- β-D-galactopyranoside) and tigogenin hexasaccharide-2 (TGHS-2, (25R)-5α-spirostan-3β-yl 4-O-[2-O-[3-O- (β-D-glucopyranosyl)-β-D-glucopyranosyl]-3-O-[4-O-(α-L-rhamnopyranosyl)-β-D-glucopyranosyl]-β-D-glucopyranosyl]- β-D-galactopyranoside), were isolated from the fresh bulbs of Camassia cusickii. In murine leukemic L1210 cells, both compounds showed cytotoxicity with an EC50 value of 0.06 µM. The morphological observation revealed that TGHS-1 and TGHS-2 induced shrinkage in cell soma and chromatin condensation, suggesting apoptotic cell death. The cell death was confirmed to be apoptosis by Annexin V binding to phosphatidylserine in the cell membrane and excluding propidium iodide. A typical apoptotic DNA ladder and the cleavage of caspase-3 were observed after treatment with TGHS-1 and TGHS-2. In the presence of both the compounds, cells with sub-G1 DNA content were detected by flow cytometric analysis, indicating that TGHS-1 and TGHS-2 (each EC50 value of 0.1 µM) are the most powerful apoptotic saponins known. These results suggest that TGHS-1 and TGHS-2 induce apoptotic cell death through caspase-3 activation.Key words: steroidal saponin, tigogenin hexasaccharide, apoptosis, DNA fragmentation, murine leukemic L1210 cells.

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