scholarly journals Quercetin Induces Mitochondrial Mediated Apoptosis and Protective Autophagy in Human Glioblastoma U373MG Cells

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Hyeonji Kim ◽  
Jeong Yong Moon ◽  
Kwang Seok Ahn ◽  
Somi Kim Cho
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Dependent Manner ◽  
Antitumor Effects ◽  
Concentration Dependent Manner ◽  
Proteolytic Activation ◽  
Caspase 7 ◽  
Dietary Flavonoid ◽  
Malignant Glioma Cells

Quercetin is a dietary flavonoid with known antitumor effects against several types of cancers by promoting apoptotic cell death and inducing cell cycle arrest. However, U373MG malignant glioma cells expressing mutant p53 are resistant to a 24 h quercetin treatment. In this study, the anticancer effect of quercetin was reevaluated in U373MG cells, and quercetin was found to be significantly effective in inhibiting proliferation of U373MG cells in a concentration-dependent manner after 48 and 72 h of incubation. Quercetin induced U373MG cell death through apoptosis, as evidenced by the increased number of cells in the sub-G1 phase, the appearance of fragmented nuclei, decreased mitochondrial membrane potential, proteolytic activation of caspase-3 and caspase-7, an increase in caspase-3 and 9 activities, and degradation of poly(ADP-ribose) polymerase protein. Furthermore, quercetin activated JNK and increased the expression of p53, which translocated to the mitochondria and simultaneously led to the release of cytochrome c from mitochondria to the cytosol. We also found that quercetin induced autophagy. Pretreatment with chloroquine, an autophagy inhibitor, strongly augmented apoptosis in U373MG cells, indicating that quercetin induced protective autopagy in U373MG cells.

scholarly journals Cytoprotective Activity ofGlycyrrhizae radixExtract against Arsenite-Induced Cytotoxicity

2008 ◽  
Vol 5 (2) ◽  
pp. 165-171 ◽  
Author(s):  
Sang Chan Kim ◽  
Sook Jahr Park ◽  
Jong Rok Lee ◽  
Jung Cheol Seo ◽  
Chae Ha Yang ◽  
...  
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Herbal Medicines ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Cytoprotective Activity ◽  
Flow Cytometric ◽  
H4iie Cells ◽  
Cell Line Cell

Licorice,Glycyrrhizae radix, is one of the herbal medicines in East Asia that has been commonly used for treating various diseases, including stomach disorders. This study investigated the effect of licorice on arsenite (As)-induced cytotoxicity in H4IIE cells, a rat hepatocyte-derived cell line. Cell viability was significantly diminished in As-treated H4IIE cells in a time and concentration-dependent manner. Furthermore, results from flow cytometric assay and DNA laddering in H4IIE cells showed that As treatment induced apoptotic cell death by activating caspase-3. Licorice (0.1 and 1.0 mg ml−1) treatment significantly inhibited cell death and the activity of caspase-3 in response to As exposure. These results demonstrate that licorice induced a cytoprotective effect against As-induced cell death by inhibition of caspase-3.

scholarly journals Anticancer effects of mifepristone on human uveal melanoma cells

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Prisca Bustamante Alvarez ◽  
Alexander Laskaris ◽  
Alicia A. Goyeneche ◽  
Yunxi Chen ◽  
Carlos M. Telleria ◽  
...  
Keyword(s):  
Cell Death ◽  
Cell Growth ◽  
Progesterone Receptor ◽  
Dna Fragmentation ◽  
Cell Lines ◽  
Caspase 3 ◽  
Annexin V ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Free Dna

Abstract Background Uveal melanoma (UM), the most prevalent intraocular tumor in adults, is a highly metastatic and drug resistant lesion. Recent studies have demonstrated cytotoxic and anti-metastatic effects of the antiprogestin and antiglucocorticoid mifepristone (MF) in vitro and in clinical trials involving meningioma, colon, breast, and ovarian cancers. Drug repurposing is a cost-effective approach to bring approved drugs with good safety profiles to the clinic. This current study assessed the cytotoxic effects of MF in human UM cell lines of different genetic backgrounds. Methods The effects of incremental concentrations of MF (0, 5, 10, 20, or 40 μM) on a panel of human UM primary (MEL270, 92.1, MP41, and MP46) and metastatic (OMM2.5) cells were evaluated. Cells were incubated with MF for up to 72 h before subsequent assays were conducted. Cellular functionality and viability were assessed by Cell Counting Kit-8, trypan blue exclusion assay, and quantitative label-free IncuCyte live-cell analysis. Cell death was analyzed by binding of Annexin V-FITC and/or PI, caspase-3/7 activity, and DNA fragmentation. Additionally, the release of cell-free DNA was assessed by droplet digital PCR, while the expression of progesterone and glucocorticoid receptors was determined by quantitative real-time reverse transcriptase PCR. Results MF treatment reduced cellular proliferation and viability of all UM cell lines studied in a concentration-dependent manner. A reduction in cell growth was observed at lower concentrations of MF, with evidence of cell death at higher concentrations. A significant increase in Annexin V-FITC and PI double positive cells, caspase-3/7 activity, DNA fragmentation, and cell-free DNA release suggests potent cytotoxicity of MF. None of the tested human UM cells expressed the classical progesterone receptor in the absence or presence of MF treatment, suggesting a mechanism independent of the modulation of the cognate nuclear progesterone receptor. In turn, all cells expressed non-classical progesterone receptors and the glucocorticoid receptor. Conclusion This study demonstrates that MF impedes the proliferation of UM cells in a concentration-dependent manner. We report that MF treatment at lower concentrations results in cell growth arrest, while increasing the concentration leads to lethality. MF, which has a good safety profile, could be a reliable adjuvant of a repurposing therapy against UM.

scholarly journals Antiviral effect of dehydroepiandrosterone on Japanese encephalitis virus infection

2005 ◽  
Vol 86 (9) ◽  
pp. 2513-2523 ◽  
Author(s):  
Chia-Che Chang ◽  
Yen-Chuan Ou ◽  
Shue-Ling Raung ◽  
Chun-Jung Chen
Keyword(s):  
Cell Death ◽  
Japanese Encephalitis Virus ◽  
Japanese Encephalitis ◽  
Apoptotic Cell ◽  
Antiviral Effect ◽  
Encephalitis Virus ◽  
Apoptotic Cell Death ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Japanese Encephalitis Virus Infection

Japanese encephalitis virus (JEV), which causes neurological disorders, completes its life cycle and triggers apoptotic cell death in infected cells. Dehydroepiandrosterone (DHEA), an adrenal-derived steroid, has been implicated in protection against neurotoxicity and protection of animals from viral-induced encephalitis, resulting in an increased survival rate of the animals. Currently, the mechanisms underlying the beneficial effects of DHEA against the virus are largely unknown. In this study, DHEA suppression of JEV replication and virus-induced apoptosis in murine neuroblastoma (N18) cells was investigated. It was found that DHEA suppressed JEV-induced cytopathic effects, JEV-induced apoptotic cell death and JEV propagation in a concentration-dependent manner. Antiviral activity was more efficient in cultures treated with DHEA immediately after viral adsorption compared with that in cultures receiving delayed administration after adsorption or transient exposure before adsorption. JEV-induced cytotoxicity was accompanied by the inactivation of extracellular signal-regulated protein kinase (ERK). Inactivation of ERK by JEV infection was reversed by DHEA. When cells were treated with the ERK inhibitor U0126, DHEA lost its antiviral effect. Activation of ERK by anisomycin mimicked the action of DHEA in suppressing JEV-induced cytotoxicity. DHEA-related compounds, such as its sulfate ester (DHEAS) and pregnenolone, were unable to suppress JEV-induced cytotoxicity and ERK inactivation. The hormone-receptor antagonists ICI 182780 and flutamide failed to abrogate the antiviral effect of DHEA. These findings suggest that the antiviral effect of DHEA is not linked directly to the genomic steroid-receptor pathways and suggest that the signalling pathways of ERK play a role in the antiviral action of DHEA.

Failure of gelsolin overexpression to regulate lymphocyte apoptosis

Blood ◽  
2000 ◽  
Vol 95 (11) ◽  
pp. 3483-3488 ◽  
Author(s):  
S. Celeste Posey ◽  
Maria Paola Martelli ◽  
Toshifumi Azuma ◽  
David J. Kwiatkowski ◽  
Barbara E. Bierer
Keyword(s):  
Cell Death ◽  
Growth Factor ◽  
Actin Filaments ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Morphological Changes ◽  
Apoptotic Cell Death ◽  
Model Systems ◽  
Dependent Manner ◽  
Amino Terminal

Abstract The actin regulatory protein gelsolin cleaves actin filaments in a calcium- and polyphosphoinositide-dependent manner. Gelsolin has recently been described as a novel substrate of the cysteinyl protease caspase-3, an effector protease activated during apoptosis. Cleavage by caspase-3 generates an amino-terminal fragment of gelsolin that can sever actin filaments independently of calcium regulation. The disruption of the actin cytoskeleton by cleaved gelsolin is hypothesized to mediate many of the downstream morphological changes associated with apoptosis. In contrast, overexpression of full-length gelsolin has also been reported to inhibit apoptotic cell death upstream of the activation of caspase-3, suggesting that gelsolin may also act prior to commitment to cell death. The authors previously observed that actin stabilization by the cell permeant agent jasplakinolide enhanced cell death upon interleukin (IL)-2 or IL-3 withdrawal from growth-factor–dependent lymphocyte cell lines, and hypothesized that actin polymerization could alter the activity of gelsolin, thus enhancing apoptosis. Here the authors show that constitutive overexpression of gelsolin did not, however, inhibit or dramatically enhance apoptotic cell death upon growth-factor withdrawal, nor did it modify sensitivity to jasplakinolide. In contrast to previous reports, overexpression of gelsolin in Jurkat T cells did not prevent or delay apoptosis induced by Fas ligation or ceramide treatment. Overexpressed gelsolin protein was cleaved during apoptosis, as seen previously in this and other cell types. In these model systems, therefore, the level of gelsolin expression was not a rate-limiting determinant in commitment to or time to the morphological changes of apoptosis.

A Newly Developed IAP Inhibitor (IAPi) Induces Apoptosis of Human Myeloma Cells and Synergises with Conventional and Novel Anti-Myeloma Therapeutics

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 5156-5156
Author(s):  
Tiffany T. Khong ◽  
Andrew Spencer
Keyword(s):  
Bone Marrow ◽  
Cell Death ◽  
Caspase 3 ◽  
Mononuclear Cells ◽  
Nuclear Translocation ◽  
Receptor Activation ◽  
Cell Killing ◽  
Parp Cleavage ◽  
Dependent Manner ◽  
Caspase 7

Abstract The IAPs (inhibitor of apoptosis proteins) are a family of 8 structurally related caspase inhibitors that efficiently block the execution phase of apoptosis induced by various stimuli including death receptor activation. Based on reports of elevated expression of IAPs in multiple myeloma (MM) we evaluated the therapeutic potential of a novel IAP (IAPi) in MM. A panel of 9 genetically heterogeneous human myeloma cell lines (HMCL) were screened and found to be sensitive to IAPi with an IC50 of 25 to 50μM in a dose and time dependent manner. Induction of cell death was confirmed by Annexin V and propidium iodide staining of treated HMCL. Mechanistic studies were then undertaken utilising 3 HMCL with high (NCI H929), intermediate (LP-1) or low (OPM2) sensitivity to IAPi with QRT-PCR demonstrating the highest expression of XIAP in the most IAPi sensitive HMCL NCI H929. In all 3 HMCL IAPi exposure was associated with rapid cleavage of caspase 3, caspase 7, ROCK-1, cytosolic degradation of ICAD, nuclear translocation of CAD and PARP cleavage confirming cell death via apoptosis. Replicate experiments utilising caspase specific inhibitors and siRNA knock-down of caspases 3 and 7 confirmed this to be both caspase 3 and caspase 7 dependent. Co-culture of HMCL with conditioned media from the human stromal cell line HS-5 or primary MM bone marrow mononuclear cells reproducibly demonstrated enhanced IAPi-induced HMCL apoptosis with a median increase in HMCL killing of 1.2 fold compared to HMCL treated in standard culture media. Similarly, IAPi activity was not abrogated but enhanced in the presence of either IL-6 or IGF-1 (1.1 and 1.3 fold, respectively) when compared to HMCL treated without the addition of exogenous growth factors. Primary MM cells from 9 multiply relapsed MM patients were treated with IAPi in an autologous bone marrow co-culture assay with concentrations of IAPi ranging from 10–100μM inducing a median of 23% (range, 8–56% as determined by Apo2.7 staining) MM cell killing at 48 hours. Finally the anti-MM activity of IAPi in combination with other conventional (etoposide, adriamycin, velcade) and novel (HSP90 inhibitor, agonistic TRAIL DR5 monoclonal antibody) therapeutics was investigated against both HMCL and primary MM tumour samples. All combinations showed synergistic cell killing with Combination Indices of <1 (Calcusyn) when compared to either agent alone. We conclude that IAPi induces apoptosis of MM cells via a caspase 3 and caspase 7 dependent process. Furthermore, IAPi retains anti-MM activity in the context of pro-survival cytokine exposure and induces synergistic killing of MM cells when combined with both conventional and novel anti-MM therapeutics. Based on these preliminary observations further investigation of the role of IAPi as a potential tumour sensitising agent for the therapy of MM is justified.

scholarly journals Mechanistic Interplay Between Autophagy and Apoptotic Signaling in Endosulfan-Induced Dopaminergic Neurotoxicity: Relevance to the Adverse Outcome Pathway in Pesticide Neurotoxicity

2019 ◽  
Vol 169 (2) ◽  
pp. 333-352 ◽  
Author(s):  
Chunjuan Song ◽  
Adhithiya Charli ◽  
Jie Luo ◽  
Zainab Riaz ◽  
Huajun Jin ◽  
...  
Keyword(s):  
Cell Death ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Neuronal Cells ◽  
Apoptotic Cell Death ◽  
Neuronal Cultures ◽  
Adverse Outcome Pathway ◽  
Proteolytic Activation ◽  
Apoptotic Signaling ◽  
Caspase 2

Abstract Chronic exposure to pesticides is implicated in the etiopathogenesis of Parkinson’s disease (PD). Previously, we showed that dieldrin induces dopaminergic neurotoxicity by activating a cascade of apoptotic signaling pathways in experimental models of PD. Here, we systematically investigated endosulfan’s effect on the interplay between apoptosis and autophagy in dopaminergic neuronal cell models of PD. Exposing N27 dopaminergic neuronal cells to endosulfan rapidly induced autophagy, indicated by an increased number of autophagosomes and LC3-II accumulation. Prolonged endosulfan exposure (>9 h) triggered apoptotic signaling, including caspase-2 and -3 activation and protein kinase C delta (PKCδ) proteolytic activation, ultimately leading to cell death, thus demonstrating that autophagy precedes apoptosis during endosulfan neurotoxicity. Furthermore, inhibiting autophagy with wortmannin, a phosphoinositide 3-kinase inhibitor, potentiated endosulfan-induced apoptosis, suggesting that autophagy is an early protective response against endosulfan. Additionally, Beclin-1, a major regulator of autophagy, was cleaved during the initiation of apoptotic cell death, and the cleavage was predominantly mediated by caspase-2. Also, caspase-2 and caspase-3 inhibitors effectively blocked endosulfan-induced apoptotic cell death. CRISPR/Cas9-based stable knockdown of PKCδ significantly attenuated endosulfan-induced caspase-3 activation, indicating that the kinase serves as a regulatory switch for apoptosis. Additional studies in primary mesencephalic neuronal cultures confirmed endosulfan’s effect on autophagy and neuronal degeneration. Collectively, our results demonstrate that a functional interplay between autophagy and apoptosis dictate pesticide-induced neurodegenerative processes in dopaminergic neuronal cells. Our study provides insight into cell death mechanisms in environmentally linked neurodegenerative diseases.

Failure of gelsolin overexpression to regulate lymphocyte apoptosis

Blood ◽  
2000 ◽  
Vol 95 (11) ◽  
pp. 3483-3488
Author(s):  
S. Celeste Posey ◽  
Maria Paola Martelli ◽  
Toshifumi Azuma ◽  
David J. Kwiatkowski ◽  
Barbara E. Bierer
Keyword(s):  
Cell Death ◽  
Growth Factor ◽  
Actin Filaments ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Morphological Changes ◽  
Apoptotic Cell Death ◽  
Model Systems ◽  
Dependent Manner ◽  
Amino Terminal

The actin regulatory protein gelsolin cleaves actin filaments in a calcium- and polyphosphoinositide-dependent manner. Gelsolin has recently been described as a novel substrate of the cysteinyl protease caspase-3, an effector protease activated during apoptosis. Cleavage by caspase-3 generates an amino-terminal fragment of gelsolin that can sever actin filaments independently of calcium regulation. The disruption of the actin cytoskeleton by cleaved gelsolin is hypothesized to mediate many of the downstream morphological changes associated with apoptosis. In contrast, overexpression of full-length gelsolin has also been reported to inhibit apoptotic cell death upstream of the activation of caspase-3, suggesting that gelsolin may also act prior to commitment to cell death. The authors previously observed that actin stabilization by the cell permeant agent jasplakinolide enhanced cell death upon interleukin (IL)-2 or IL-3 withdrawal from growth-factor–dependent lymphocyte cell lines, and hypothesized that actin polymerization could alter the activity of gelsolin, thus enhancing apoptosis. Here the authors show that constitutive overexpression of gelsolin did not, however, inhibit or dramatically enhance apoptotic cell death upon growth-factor withdrawal, nor did it modify sensitivity to jasplakinolide. In contrast to previous reports, overexpression of gelsolin in Jurkat T cells did not prevent or delay apoptosis induced by Fas ligation or ceramide treatment. Overexpressed gelsolin protein was cleaved during apoptosis, as seen previously in this and other cell types. In these model systems, therefore, the level of gelsolin expression was not a rate-limiting determinant in commitment to or time to the morphological changes of apoptosis.

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