ASK1-P38 Pathway is Important for Anoikis Induced by Microtubule-Targeting Aryl Chloroethylureas

PURPOSE. We investigated the involvement of MAPK signaling in the cell death mechanisms of classical microtubule interfering agents (MIA) and aryl-3-(2-chloroethyl)ureas (CEU) acting as antimitotics, along with CEU that don’t affect directly microtubules (non-MIA CEU). METHODS. To ascertain the activated signaling pathway profile of MIA and non-MIA CEU, Western blot, immunoprecipitation and transfection experiments were performed. RESULTS. Non-MIA CEU do not activate p38, as opposed to MIA, and the extent of ERK and JNK activation is lower than in response to MIA. The effect of MIA and non-MIA CEU on focal adhesion associated protein was also studied; MIA were shown to induce focal adhesion dismantlement associated with a sustained increase in paxillin phosphorylation and FAK cleavage, as opposed to non-MIA CEU. In addition, bcl-2 phosphorylation and AKT cleavage, induced by all MIA tested, was not observed in response to non-MIA CEU further emphasizing the differential cell death mechanisms induced by MIA and non-MIA CEU. Pharmacologic and genetic approaches emphasize that the ASK1-p38 pathway activation contributes to the cytotoxic mechanism of MIA, in contrast to non-MIA CEU. ASK1-p38 is important for increased paxillin phosphorylation and FAK cleavage, suggesting that ASK-1-p38 is an upstream event of FA structure dismantlement induced by MIA. Moreover, the endogen inhibitor of ASK-1, thioredoxin, is released from ASK-1 in response to MIA as opposed to non-MIA CEU. CONCLUSION. Our study supports that ASK1-p38 activation is an important signaling event, induced by MIA, which impairs focal adhesion structure and induces anchorage-dependent apoptosis or anoikis.

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Early Phosphatidylinositol 3-Kinase/Akt Pathway Activation Limits Poliovirus-Induced JNK-Mediated Cell Death

Journal of Virology ◽

10.1128/jvi.02020-07 ◽

2008 ◽

Vol 82 (7) ◽

pp. 3796-3802 ◽

Cited By ~ 33

Author(s):

Arnaud Autret ◽

Sandra Martin-Latil ◽

Cynthia Brisac ◽

Laurence Mousson ◽

Florence Colbère-Garapin ◽

...

Keyword(s):

Cell Death ◽

Tissue Injury ◽

Neuroblastoma Cells ◽

Phosphatidylinositol 3 Kinase ◽

Pathway Activation ◽

Survival Pathway ◽

The Central Nervous System ◽

Induced Apoptosis ◽

Jnk Activation ◽

Phosphatidylinositol 3

ABSTRACT Poliovirus (PV)-induced apoptosis seems to play a major role in tissue injury in the central nervous system (CNS). We have previously shown that this process involves PV-induced Bax-dependent mitochondrial dysfunction mediated by early JNK activation in IMR5 neuroblastoma cells. We showed here that PV simultaneously activates the phosphatidylinositol 3-kinase (PI3K)/Akt survival signaling pathway in these cells, limiting the extent of JNK activation and thereby cell death. JNK inhibition is associated with PI3K-dependent negative regulation of the apoptosis signal-regulating kinase 1, which acts upstream from JNK in PV-infected IMR5 cells. In poliomyelitis, this survival pathway may limit the spread of PV-induced damage in the CNS.

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Tissue Inhibitor of Metalloproteinase-1 Protects Human Breast Epithelial Cells Against Intrinsic Apoptotic Cell Death via the Focal Adhesion Kinase/Phosphatidylinositol 3-Kinase and MAPK Signaling Pathway

Journal of Biological Chemistry ◽

10.1074/jbc.m302999200 ◽

2003 ◽

Vol 278 (41) ◽

pp. 40364-40372 ◽

Cited By ~ 123

Author(s):

Xu-Wen Liu ◽

M. Margarida Bernardo ◽

Rafael Fridman ◽

Hyeong-Reh Choi Kim

Keyword(s):

Cell Death ◽

Epithelial Cells ◽

Focal Adhesion ◽

Apoptotic Cell ◽

Mapk Signaling ◽

Tissue Inhibitor Of Metalloproteinase ◽

Human Breast ◽

Phosphatidylinositol 3 Kinase ◽

Breast Epithelial Cells ◽

Human Breast Epithelial Cells

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Antiprolactinoma Effect of Hordenine by Inhibiting MAPK Signaling Pathway Activation in Rats

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2020/3107290 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9

Author(s):

Xiong Wang ◽

Run-zhu Guo ◽

Li Ma ◽

Qiao-yan Ding ◽

Jun-hua Meng ◽

...

Keyword(s):

Pituitary Gland ◽

Signaling Pathway ◽

Mapk Pathway ◽

Interleukin 1 ◽

Mapk Signaling ◽

Mapk Signaling Pathway ◽

Mitogen Activated Protein Kinase ◽

Pathway Activation ◽

Mitogen Activated Protein ◽

Jnk Activation

Prolactinomas are harmful to human health, and the clinical first-line treatment drug is bromocriptine. However, 20% prolactinomas patients did not respond to bromocriptine. Hordenine is an alkaloid separated from Fructus Hordei Germinatus, which showed significant antihyperprolactinemia activity in rats. The aim of this study was to explore the effect and mechanism of hordenine on prolactinomas in rats. The study used estradiol to induce prolactinomas, which caused the activation of the pituitary mitogen-activated protein kinase (MAPK) pathway in rats significantly. The treatment of hordenine restored estradiol, induced the overgrowth of pituitary gland, and reduced the prolactin (PRL) accumulation in the serum and pituitary gland of rats by blocking the MAPK (p38, ERK1/2, and JNK) activation and production of inflammatory cytokines, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). The antiprolactinoma effect of hordenine was mediated by inhibiting the MAPK signaling pathway activation in rats.

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Combining RAD001 (Everolimus) with Proteasome Inhibitors Bortezomib (Velcade) or MG132 Significantly Enhances Pre-B ALL Cell Death in Vitro.

Blood ◽

10.1182/blood.v114.22.2743.2743 ◽

2009 ◽

Vol 114 (22) ◽

pp. 2743-2743

Author(s):

Philip O. Saunders ◽

Kenneth F Bradstock ◽

Linda J. Bendall

Keyword(s):

Cell Death ◽

Lymphoblastic Leukemia ◽

Significant Proportion ◽

Receptor Activation ◽

Jnk Pathway ◽

Dna Damaging Agents ◽

Pathway Activation ◽

Wide Range ◽

Jnk Activation

Abstract Abstract 2743 Poster Board II-719 Mammalian target of rapamycin (mTOR) inhibitors have shown potential as novel therapeutic agents with efficacy against a wide range of tumors including precursor-B acute lymphoblastic leukemia (pre-B ALL). We have previously reported RAD001 (16μM) induces JNK pathway activation in pre-B ALL cells and that combining RAD001 with DNA damaging agents in vitro significantly enhanced JNK dependent death, via a caspase dependent mechanism. We sought to evaluate agents, which may favor JNK activation and promote JNK dependent cell death in pre-B ALL cells. Bortezomib and MG132 have both been reported to activate the JNK pathway via death receptor activation, with reported efficacy in pre-B ALL. Consistent with the literature we observed enhanced JNK pathway activation in pre-B ALL cells treated with bortezomib. Analysis of annexin V and 7AAD expression by flow cytometry utilizing JNK inhibitor SP600125 (5μM) showed that a significant proportion of pre-B ALL cell death observed with bortezomib was JNK dependent. Combining RAD001 (4–16μM) with bortezomib in vitro (10–20nM) significantly enhanced cell death in pre-B ALL cell lines and patient cases at 24 hours. This observation was supported by equivalent observations combining MG132 (250–500nM) with RAD001 (8–16μM). The degree of enhanced killing was greater than that achieved combining RAD001 (16μM) with DNA damaging agents. Enhanced killing was also achieved at a significantly lower dose of RAD001 relative to combination therapy with DNA damage. Utilizing JNK inhibitor SP600125 (5μM) we determined that a significant proportion of enhanced killing was JNK dependent. In conclusion we have identified two novel and clinically available agents which when combined can significantly enhance pre-B ALL cell death. Our observations suggest combining agents which induce JNK activation has the potential to enhance clinical responses in pre-B ALL, particularly for patients with advanced or relapsed disease, for whom treatment with cytotoxic chemotherapy offers little hope of improved survival. In vivo studies will provide further insight into this promising strategy. Disclosures: No relevant conflicts of interest to declare.

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Cytotoxic Mechanism for Silver Nanoparticles Based High-Content Cellomics and Transcriptome Sequencing

Journal of Biomedical Nanotechnology ◽

10.1166/jbn.2019.2785 ◽

2019 ◽

Vol 15 (7) ◽

pp. 1401-1414 ◽

Cited By ~ 3

Author(s):

Yan Huang ◽

Xiaoying Lü ◽

Xiaoqiang Lü

Keyword(s):

Signaling Pathway ◽

Focal Adhesion ◽

Transcriptome Sequencing ◽

Mapk Signaling ◽

Mapk Signaling Pathway ◽

Differentially Expressed ◽

Atp Content ◽

Biological Pathway Analysis ◽

Key Genes ◽

Cytotoxic Mechanism

The aim of this study was to investigate the toxic mechanism for differently sized silver nanoparticles (SNPs) on human dermal fibroblasts (HDFs), by combining high content cellomics and transcriptome sequencing. First, the influences of five SNPs (SNP-5, SNP-20, SNP-50, SNP-100, and SNP-200) on O–2, focal adhesion, cytoskeleton and ATP content in HDFs were studied with high content screening and colorimetric method, and the role to cytotoxicity was analysed. Transcriptome sequencing technique was then to filter differentially expressed genes induced by SNPs after 4 h treatment. Key pathways in SNP-induced cytotoxicity were also screened via biological pathway analysis. Furthermore, key genes in HDFs after SNP-induced cytotoxicity were determined through matching analysis with previously obtained important microRNAs and their expression levels were verified with qRT-PCR. Cytological experiments showed that the SNP-5 had the strongest effects on O–2, focal adhesion, cytoskeleton and ATP content, while SNP-20 had the smallest effects. Transcriptome sequencing results showed that 3848, 4213, 2999, 3251 and 5104 genes were found to be differentially expressed in HDFs after treatment with five SNPs. Biological pathway analysis for 1643 uniformly differentially expressed genes revealed that MAPK signaling pathway was the key pathway in SNP-induced cytotoxicity. Two key genes, SOS1 and CDC25B, which are involved in MAPK signaling pathway were finally identified through matching analysis with important microRNAs and verification. In conclusion, the cytotoxic mechanism for SNPs induced cytotoxicity in HDFs involved SNPs down-regulated expression of SOS1 and CDC25B through miR-424-5p in the key MAPK signaling pathway, through blocking of cell cycle, promotion of apoptosis, ultimately leading to cytotoxicity.

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The JNK Pathway Is a Significant Determinant of Sensitivity to DNA Damaging Agents in Pre-B ALL.

Blood ◽

10.1182/blood.v114.22.3786.3786 ◽

2009 ◽

Vol 114 (22) ◽

pp. 3786-3786

Author(s):

Philip O. Saunders ◽

Kenneth F Bradstock ◽

Linda J. Bendall

Keyword(s):

Cell Cycle ◽

Dna Damage ◽

Cell Death ◽

Dna Damage Response ◽

Genotoxic Stress ◽

Jnk Pathway ◽

Dna Damaging Agents ◽

Pathway Activation ◽

Damage Response ◽

Jnk Activation

Abstract Abstract 3786 Poster Board III-722 The JNK pathway is reported to facilitate AP1 binding and promote apoptosis depending on cell type and environmental conditions. We have previously reported RAD001 (16μM) induces JNK pathway activation in pre-B ALL cells. We sought to evaluate the impact of changes in JNK pathway activation on pre-B ALL viability in vitro. Using JNK inhibitor SP600125 titrated to inhibit c-Jun activation, we determined that cell death in pre-B ALL cells treated with RAD001 (16μM) alone was not JNK dependent. In contrast, combining RAD001 (16μM) with DNA damaging agents significantly enhanced JNK dependent death. This difference indicates that additional factors, including genotoxic stress, are required for JNK activation to induce pre-B ALL cell death. The JNK pathway is reported to suppress transcriptional activation of key mediators of the DNA damage response. We observed that JNK activation in cells treated with RAD001 (16μM) and DNA damaging agents was associated with suppression of p53 and p21 relative to DNA damage alone. This result was supported by the observation of enhanced p53 and p21 expression in pre-B ALL cells treated with DNA damaging agents in the presence of the JNK inhibitor SP600125. Analysis of DNA content and proliferation antigen expression in pre-B ALL cells treated with RAD001 (16μM) and DNA damaging agents revealed JNK activation was associated with a significant increase in the proportion of cells in S phase, relative to DNA damage alone, which caused a G1 and G2 cell cycle arrest. Further evidence that the JNK pathway impacts on the DNA damage response was provided by the observation that pre-B ALL cells treated with DNA damaging agents and JNK inhibitor SP600125 demonstrated reduced PCNA expression at G1 and G2 and reduced expression of mitotic antigen phospho-Histone–H3. This is consistent with enhanced regulation at G1-S and G2-M checkpoints. The results indicate changes in JNK pathway activation impact on the cell cycle response to DNA damage. In conclusion we have identified that the JNK pathway has a significant impact on the sensitivity of pre-B ALL cells to DNA damaging agents. JNK activation in the presence of genotoxic stress significantly enhanced pre-B ALL cell death, associated with suppression of key mediators of the DNA damage response, p53 and p21. We found that changes in JNK activation altered the cell cycle response to DNA damage. Further study is required to determine if changes in cell cycle regulation in the presence of DNA damage is causal to JNK dependent cell death. Additional studies to identify intracellular signal pathways which facilitate JNK dependent cell death are warranted. Our observations suggest combining agents which induce JNK activation with conventional chemotherapy or selected novel agents has the potential to enhance clinical responses in pre-B ALL. Disclosures: No relevant conflicts of interest to declare.

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Arctic Ground Squirrel (Spermophilus Parryii) Hippocampal Neurons Tolerate Prolonged Oxygen—Glucose Deprivation and Maintain Baseline ERK1/2 and JNK Activation Despite Drastic ATP Loss

Journal of Cerebral Blood Flow & Metabolism ◽

10.1038/jcbfm.2008.20 ◽

2008 ◽

Vol 28 (7) ◽

pp. 1307-1319 ◽

Cited By ~ 30

Author(s):

Sherri L Christian ◽

Austin P Ross ◽

Huiwen W Zhao ◽

Heidi J Kristenson ◽

Xinhua Zhan ◽

...

Keyword(s):

Cell Death ◽

Hippocampal Slices ◽

Glucose Deprivation ◽

Mapk Signaling ◽

Ground Squirrels ◽

Atp Depletion ◽

Mitogen Activated Protein Kinase ◽

Oxygen Glucose Deprivation ◽

Acute Hippocampal Slices ◽

Jnk Activation

Oxygen—glucose deprivation (OGD) initiates a cascade of intracellular responses that culminates in cell death in sensitive species. Neurons from Arctic ground squirrels (AGS), a hibernating species, tolerate OGD in vitro and global ischemia in vivo independent of temperature or torpor. Regulation of energy stores and activation of mitogen-activated protein kinase (MAPK) signaling pathways can regulate neuronal survival. We used acute hippocampal slices to investigate the role of ATP stores and extracellular signal-regulated kinase (ERK)1/2 and Jun NH2-terminal kinase (JNK) MAPKs in promoting survival. Acute hippocampal slices from AGS tolerated 30 mins of OGD and showed a small but significant increase in cell death with 2 h OGD at 37 C. This tolerance is independent of hibernation state or season. Neurons from AGS survive OGD despite rapid ATP depletion by 3 mins in interbout euthermic AGS and 10 mins in hibernating AGS. Oxygen—glucose deprivation does not induce JNK activation in AGS and baseline ERK1/2 and JNK activation is maintained even after drastic depletion of ATP. Surprisingly, inhibition of ERK1/2 or JNK during OGD had no effect on survival, whereas inhibition of JNK increased cell death during normoxia. Thus, protective mechanisms promoting tolerance to OGD by AGS are downstream from ATP loss and are independent of hibernation state or season.

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