scholarly journals Induction of Apoptotic Cell Death in Human Leukemia U937 Cells by C18 Hydroxy Unsaturated Fatty Acid Isolated from Red Alga Tricleocarpa jejuensis

Marine Drugs ◽  
2021 ◽  
Vol 19 (3) ◽  
pp. 138
Author(s):  
Shijiao Zha ◽  
Mikinori Ueno ◽  
Yan Liang ◽  
Seiji Okada ◽  
Tatsuya Oda ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Dna Fragmentation ◽  
Hydroxy Group ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Red Alga ◽  
Caspase 8 ◽  
Human Leukemia ◽  
U937 Cells ◽  
Cleavage Activity

Our previous studies have found that (±)-(E)-12-hydroxyoctadec-10-enoic acid (HOEA) isolated from the red alga Tricleocarpa jejuensis showed cytotoxic effects on various living organisms including harmful microalgae, Gram-positive bacteria, and mammalian tumor cells. Since natural products with apoptosis-inducing ability can be promising anti-cancer agents, in this study, we investigated the cytotoxic mechanism of HOEA on U937 cells focusing on apoptosis induction. HOEA showed much stronger cytotoxic and cytolytic effects on U937 cells than elaidic acid, which has similar structure but no 12-hydroxy group, suggesting that hydroxy group is important for the cytotoxicity of HOEA. HOEA induced apoptotic nuclear morphological changes, DNA fragmentation, and decrease in mitochondrial membrane potential. Furthermore, time-dependent increase in annexin V+/PI+ cell population in HOEA-treated U937 cells was detected. Among the apoptosis-related reagents, caspase-family inhibitor almost completely inhibited HOEA-induced DNA fragmentation. In the analyses using specific caspase-substrates, extremely high cleavage activity toward caspase-3/7/8 substrate was observed in HOEA-treated U937 cells, and weak activities of caspase-1 and -3 were detected. Analyses using specific caspase inhibitors suggested that caspase-3 and caspase-8 might be predominantly responsible for the cleavage activity. Activation of these caspases were also confirmed by western blotting in which significant levels of cleaved forms of caspase 3, caspase 8, and PARP were detected in HOEA-treated U937 cells. Our results suggest that HOEA is capable of inducing apoptosis in U937 cells in which caspase-3 and caspase-8 might play important roles. Since the cytotoxic effect of HOEA is not strictly specific to tumor cells, development of appropriate drug delivery system for selective tumor targeting is necessary for the clinical applications to reduce the possible side effects.

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.

scholarly journals Efficient apoptosis requires feedback amplification of upstream apoptotic signals by effector caspase-3 or -7

2019 ◽  
Vol 5 (7) ◽  
pp. eaau9433 ◽  
Author(s):  
Scott McComb ◽  
Pik Ki Chan ◽  
Anna Guinot ◽  
Holmfridur Hartmannsdottir ◽  
Silvia Jenni ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Apoptotic Cell ◽  
Caspase 8 ◽  
Human Leukemia ◽  
Double Knockout ◽  
Cytochrome C Release ◽  
Discernible Effect ◽  
Effector Caspase ◽  
Caspase 6 ◽  
Apoptotic Pathways

Apoptosis is a complex multi-step process driven by caspase-dependent proteolytic cleavage cascades. Dysregulation of apoptosis promotes tumorigenesis and limits the efficacy of chemotherapy. To assess the complex interactions among caspases during apoptosis, we disrupted caspase-8, -9, -3, -7, or -6 and combinations thereof, using CRISPR-based genome editing in living human leukemia cells. While loss of apical initiator caspase-8 or -9 partially blocked extrinsic or intrinsic apoptosis, respectively, only combined loss of caspase-3 and -7 fully inhibited both apoptotic pathways, with no discernible effect of caspase-6 deficiency alone or in combination. Caspase-3/7 double knockout cells exhibited almost complete inhibition of caspase-8 or -9 activation. Furthermore, deletion of caspase-3 and -7 decreased mitochondrial depolarization and cytochrome c release upon apoptosis activation. Thus, activation of effector caspase-3 or -7 sets off explosive feedback amplification of upstream apoptotic events, which is a key feature of apoptotic signaling essential for efficient apoptotic cell death.

Synthesis and biological activities of chaetocin and its derivatives

2012 ◽  
Vol 84 (6) ◽  
pp. 1369-1378 ◽  
Author(s):  
Mikiko Sodeoka ◽  
Kosuke Dodo ◽  
Yuou Teng ◽  
Katsuya Iuchi ◽  
Yoshitaka Hamashima ◽  
...  
Keyword(s):  
Total Synthesis ◽  
Caspase 3 ◽  
Biological Activities ◽  
Caspase 8 ◽  
Human Leukemia ◽  
Mechanistic Studies ◽  
Anticancer Activities ◽  
Histone Methyltransferases ◽  
Structure Activity ◽  
Apoptosis Inducer

Chaetocin, a natural product isolated from fungi of Chaetomium species, is a member of the epipolythiodiketopiperazines (ETPs), which have various biological activities, including cytostatic and anticancer activities. Recently, the inhibitory activity toward histone methyltransferases (HMTs) was discovered for chaetocin. We previously reported the first total synthesis of chaetocin and various derivatives. During studies on the structure–activity relationship for HMT inhibition, we found that the enantiomer of chaetocin (ent-chaetocin) is a more potent apoptosis inducer than natural chaetocin in human leukemia HL-60 cells. Mechanistic studies showed that ent-chaetocin induces apoptosis through the caspase-8/caspase-3 pathway.

scholarly journals Granzyme B Short-Circuits the Need for Caspase 8 Activity during Granule-Mediated Cytotoxic T-Lymphocyte Killing by Directly Cleaving Bid

2000 ◽  
Vol 20 (11) ◽  
pp. 3781-3794 ◽  
Author(s):  
Michele Barry ◽  
Jeffrey A. Heibein ◽  
Michael J. Pinkoski ◽  
Siow-Fong Lee ◽  
Richard W. Moyer ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Caspase 3 ◽  
Cytotoxic T Lymphocyte ◽  
Granzyme B ◽  
Serine Proteinase ◽  
Target Cells ◽  
Caspase 8 ◽  
Intact Cells ◽  
Whole Cells

ABSTRACT Cytotoxic T lymphocytes (CTL) can trigger an apoptotic signal through the Fas receptor or by the exocytosis of granzyme B and perforin. Caspase activation is an important component of both pathways. Granzyme B, a serine proteinase contained in granules, has been shown to proteolytically process and activate members of the caspase family in vitro. In order to gain an understanding of the contributions of caspases 8 and 3 during granule-induced apoptosis in intact cells, we have used target cells that either stably express the rabbitpox virus-encoded caspase inhibitor SPI-2 or are devoid of caspase 3. The overexpression of SPI-2 in target cells significantly inhibited DNA fragmentation, phosphatidylserine externalization, and mitochondrial disruption during Fas-mediated cell death. In contrast, SPI-2 expression in target cells provided no protection against granzyme-mediated apoptosis, mitochondrial collapse, or cytolysis, leading us to conclude that SPI-2-inhibited caspases are not an essential requirement for the granzyme pathway. Caspase 3-deficient MCF-7 cells were found to be resistant to CTL-mediated DNA fragmentation but not to CTL-mediated cytolysis and loss of the mitochondrial inner membrane potential. Furthermore, we demonstrate that granzyme B directly cleaves the proapoptotic molecule Bid, bypassing the need for caspase 8 activation of Bid. These results provide evidence for a two-pronged strategy for mediating target cell destruction and provide evidence of a direct link between granzyme B activity, Bid cleavage, and caspase 3 activation in whole cells.

Characterization of the Resistance Mechanisms of HL60 Leukemia Cell Derivatives to Apo2L/TRAIL.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3412-3412
Author(s):  
Jinrong Cheng ◽  
Bonnie L. Hylander ◽  
Maria R. Baer ◽  
Elizabeth A. Repasky
Keyword(s):  
Cell Surface ◽  
Tumor Cells ◽  
Surface Expression ◽  
Resistance Mechanisms ◽  
Cell Surface Expression ◽  
Caspase 8 ◽  
Human Leukemia ◽  
Resistant Clone ◽  
Antibody Treatment ◽  
Hl60 Cells

Abstract Tumor necrosis factor-related apoptosis-inducing ligand (Apo2L/TRAIL) is a recently identified member of the TNF superfamily. Recombinant Apo2L/TRAIL is a promising immunotherapeutic agent for treating malignant diseases since this molecule preferentially induces apoptosis in a variety of tumor cells with apparently little toxicity to normal cells. However, it has also been shown that some tumor cells are resistant to this molecule. We hypothesized that resistance to Apo2L/TRAIL occurs through defects in the Apo2L/TRAIL-mediated apoptotic signaling pathway. To address this hypothesis, we developed several Apo2L/TRAIL-resistant HL60 derivatives (HL60/TR) by exposure of Apo2L/TRAIL-sensitive HL60 human leukemia cells to escalating levels of Apo2L/TRAIL, followed by subcloning. Two of these resistant clones (a moderately resistant clone-R1 and a highly resistant clone-R3) were selected for further study. Molecules in the Apo2L/TRAIL-mediated apoptotic pathway of R1 and R3 cells were analyzed by Western blot analysis, flow cytometry and gene sequencing and compared to those in parental HL60 cells. In the R1 cells, the activation of caspase-8 and -10 by Apo2L/TRAIL was significantly inhibited. However, R1 cells were still sensitive to Fas agonistic monoclonal antibody treatment, indicating that the FAS-mediated apoptosis-inducing pathway was intact. In the R3 cells, caspase-8 expression was completely lost and activation of caspase-10 in response to Apo2L/TRAIL was totally inhibited; R3 cells were therefore also resistant to FAS antibody treatment. Although the total protein level of death receptors DR4 and DR5 was equal in HL60 cells and in the Apo2L/TRAIL-resistant derivatives, the cell surface levels of DR4 were significantly decreased in both R1 and R3 cells, while the surface expression level of DR5 in these two clones was comparable to that on HL60 cells. No mutation in either the DR4 or DR5 genes was found in these cells. These results suggest that defective targeting of DR4 molecules to the cell surface occurs in these Apo2L/TRAIL resistant cells. Blocking cell surface DR4 significantly attenuated the sensitivity of parental HL60 cells to Apo2L/TRAIL, indicating that cell surface expression of DR4 plays a crucial role in regulating susceptibility of tumor cells to Apo2L/TRAIL. Taken together, our results demonstrate that malignant cells can develop resistance to Apo2L/TRAIL by several different mechanisms and multiple resistance mechanisms may develop in a single tumor cell (such as R3 cells). Understanding the basis of Apo2L/TRAIL resistance will help to predict sensitivity and to develop strategies to circumvent resistance.

Kalopanaxsaponin A induces apoptosis in human leukemia U937 cells through extracellular Ca2+ influx and caspase-8 dependent pathways

2008 ◽  
Vol 46 (11) ◽  
pp. 3486-3492 ◽  
Author(s):  
Jung-Hye Choi ◽  
Heon-Woo Lee ◽  
Hee-Juhn Park ◽  
Sung-Hoon Kim ◽  
Kyung-Tae Lee
Keyword(s):  
Caspase 8 ◽  
Human Leukemia ◽  
U937 Cells

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.

Midazolam induce apoptosis in mouse leydig cells and MA-10 tumor cells through caspase-3,caspase-8 and caspase-9

2011 ◽  
Vol 28 ◽  
pp. 138
Author(s):  
E. C. So ◽  
Y. X. Lin ◽  
B. M. Huang ◽  
Y. K. Wang ◽  
K. L. Wong ◽  
...  
Keyword(s):  
Tumor Cells ◽  
Leydig Cells ◽  
Caspase 3 ◽  
Caspase 8 ◽  
Caspase 9
Sign in / Sign up

Export Citation Format

Share Document