scholarly journals Anti-bacterial and anti-viral nanchangmycin displays anti-myeloma activity by targeting Otub1 and c-Maf

2020 ◽  
Vol 11 (9) ◽  
Author(s):  
Yujia Xu ◽  
Tong Sun ◽  
Kun Zeng ◽  
Min Xu ◽  
Jinhao Chen ◽  
...  
Keyword(s):  
Cell Apoptosis ◽  
Caspase 3 ◽  
Plasma Cells ◽  
Mrna Level ◽  
Annexin V ◽  
Safety And Efficacy ◽  
In Vivo Assays ◽  
Subsequent Degradation ◽  
Factor C

Abstract As a deubiqutinase Otub1 stabilizes and promotes the oncogenic activity of the transcription factor c-Maf in multiple myeloma (MM), a malignancy of plasma cells. In the screen for bioactive inhibitors of the Otub1/c-Maf axis for MM treatment, nanchangmycin (Nam), a polyketide antibiotic, was identified to suppress c-Maf activity in the presence of Otub1. By suppressing Otub1, Nam induces c-Maf polyubiquitination and subsequent degradation in proteasomes but does not alter its mRNA level. Consistently, Nam downregulates the expression of CCND2, ARK5, and ITGB7, the downstream genes regulated by c-Maf, and promotes MM cell apoptosis as evidenced by PARP and Caspase-3 cleavage, as well as Annexin V staining. In line with the hypothesis, overexpression of Otub1 partly rescues Nam-induced MM cell apoptosis, and interestingly, when Otub1 is knocked down, Nam-decreased MM cell survival is also partly ablated, suggesting Otub1 is essential for Nam anti-MM activity. Nam also displays potent anti-MM activity synergistically with Doxorubicin or lenalidomide. In the in vivo assays, Nam almost completely suppresses the growth of MM xenografts in nude mice at low dosages but it shows no toxicity. Given its safety and efficacy, Nam has a potential for MM treatment by targeting the Otub1/c-Maf axis.

scholarly journals Inhibition of the Otub1/c-Maf axis by the herbal acevaltrate induces myeloma cell apoptosis

2020 ◽  
Author(s):  
Tong Sun ◽  
Yujia Xu ◽  
Zhuan Xu ◽  
Biyin Cao ◽  
Zubin Zhang ◽  
...  
Keyword(s):  
Cell Apoptosis ◽  
Target Genes ◽  
Annexin V ◽  
Myeloma Cell ◽  
Luciferase Assay ◽  
Growth And Survival ◽  
Protein Ubiquitination ◽  
Factor C

Abstract Background : The oncogenic transcript factor c-Maf is stabilized by the deubiquitinase Otub1 and promotes myeloma cell proliferation and confers to chemoresistance. Inhibition of the Otub1/c-Maf axis is a promising therapeutic target, but there are no inhibitors reported on this specific axis. Methods: A luciferase assay was applied to screen potential inhibitors of the Otub1/c-Maf axis. Annexin V staining/flow cytometry was applied to evaluate cell apoptosis. Immunoprecipitation was applied to examine protein ubiquitination and interaction. Xenograft models in nude mice were used to evaluate anti-myeloma activity. Results: acevaltrate (AVT), isolated from Valeriana glechomifolia, was identified based on a bioactive screen against Otub1/c-Maf/luciferase. AVT disrupts the interaction of Otub1/c-Maf thus inhibiting Otub1 activity and leading to c-Maf polyubiquitination and subsequent degradation in proteasomes. Consistently, AVT inhibits c-Maf transcriptional activity and downregulates the expression of its target genes key for myeloma growth and survival. Moreover, AVT displays potent anti-myeloma activity by triggering myeloma cell apoptosis in vitro and impairing myeloma xenograft growth in vivo but presents no marked toxicity. Conclusions: The natural product AVT inhibits the Otub1/c-Maf axis and displays potent anti-myeloma activity. Given its great safety and efficacy, AVT could be further developed for MM treatment.

scholarly journals Inhibition of the Otub1/c-Maf axis by the herbal acevaltrate induces myeloma cell apoptosis

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Tong Sun ◽  
Yujia Xu ◽  
Zhuan Xu ◽  
Biyin Cao ◽  
Zubin Zhang ◽  
...  
Keyword(s):  
Cell Apoptosis ◽  
Target Genes ◽  
Annexin V ◽  
Myeloma Cell ◽  
Luciferase Assay ◽  
Growth And Survival ◽  
Protein Ubiquitination ◽  
Factor C

Abstract Background The oncogenic transcript factor c-Maf is stabilized by the deubiquitinase Otub1 and promotes myeloma cell proliferation and confers to chemoresistance. Inhibition of the Otub1/c-Maf axis is a promising therapeutic target, but there are no inhibitors reported on this specific axis. Methods A luciferase assay was applied to screen potential inhibitors of Otub1/c-Maf. Annexin V staining/flow cytometry was applied to evaluate cell apoptosis. Immunoprecipitation was applied to examine protein ubiquitination and interaction. Xenograft models in nude mice were used to evaluate anti-myeloma activity of AVT. Results Acevaltrate (AVT), isolated from Valeriana glechomifolia, was identified based on a bioactive screen against the Otub1/c-Maf/luciferase system. AVT disrupts the interaction of Otub1/c-Maf thus inhibiting Otub1 activity and leading to c-Maf polyubiquitination and subsequent degradation in proteasomes. Consistently, AVT inhibits c-Maf transcriptional activity and downregulates the expression of its target genes key for myeloma growth and survival. Moreover, AVT displays potent anti-myeloma activity by triggering myeloma cell apoptosis in vitro and impairing myeloma xenograft growth in vivo but presents no marked toxicity. Conclusions The natural product AVT inhibits the Otub1/c-Maf axis and displays potent anti-myeloma activity. Given its great safety and efficacy, AVT could be further developed for MM treatment.

Annexin V decreases PS-mediated macrophage efferocytosis and deteriorates elastase-induced pulmonary emphysema in mice

2012 ◽  
Vol 303 (10) ◽  
pp. L852-L860 ◽  
Author(s):  
S. Yoshida ◽  
N. Minematsu ◽  
S. Chubachi ◽  
H. Nakamura ◽  
M. Miyazaki ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Cell Apoptosis ◽  
Pulmonary Emphysema ◽  
Keratinocyte Growth Factor ◽  
Annexin V ◽  
Phenotypic Change ◽  
Chronic Obstructive ◽  
Mrna Levels

Efferocytosis is believed to be a key regulator for lung inflammation in chronic obstructive pulmonary disease. In this study we pharmacologically inhibited efferocytosis with annexin V and attempted to determine its impact on the progression of pulmonary emphysema in mouse. We first demonstrated in vitro and in vivo efferocytosis experiments using annexin V, an inhibitor for phosphatidylserine-mediated efferocytosis. We then inhibited efferocytosis in porcine pancreatic elastase (PPE)-treated mice. PPE-treated mice were instilled annexin V intranasally starting from day 8 until day 20. Mean linear intercept (Lm) was measured, and cell apoptosis was assessed in lung specimen obtained on day 21. Cell profile, apoptosis, and mRNA expression of matrix metalloproteinases (MMPs) and growth factors were evaluated in bronchoalveolar lavage (BAL) cells on day 15. Annexin V attenuated macrophage efferocytosis both in vitro and in vivo. PPE-treated mice had a significant higher Lm, and annexin V further increased that by 32%. More number of macrophages was found in BAL fluid in this group. Interestingly, cell apoptosis was not increased by annexin V treatment both in lung specimens and BAL fluid, but macrophages from mice treated with both PPE and annexin V expressed higher MMP-2 mRNA levels and had a trend for higher MMP-12 mRNA expression. mRNA expression of keratinocyte growth factor tended to be downregulated. We showed that inhibited efferocytosis with annexin V worsened elastase-induced pulmonary emphysema in mice, which was, at least partly, attributed to a lack of phenotypic change in macrophages toward anti-inflammatory one.

Crambene induces pancreatic acinar cell apoptosis via the activation of mitochondrial pathway

2006 ◽  
Vol 291 (1) ◽  
pp. G95-G101 ◽  
Author(s):  
Yang Cao ◽  
Sharmila Adhikari ◽  
Abel Damien Ang ◽  
Marie Véronique Clément ◽  
Matthew Wallig ◽  
...  
Keyword(s):  
Acinar Cell ◽  
Cell Apoptosis ◽  
Caspase 3 ◽  
Fas Ligand ◽  
Annexin V ◽  
Acinar Cells ◽  
Mitochondrial Pathway ◽  
Pancreatic Acinar Cell ◽  
Pancreatic Acinar Cells ◽  
Pancreatic Acini

We investigated the apoptotic pathway activated by crambene (1-cyano-2-hydroxy-3-butene), a plant nitrile, on pancreatic acinar cells. As evidenced by annexin V-FITC staining, crambene treatment for 3 h induced the apoptosis but not necrosis of pancreatic acini. Caspase-3, -8, and -9 activities in acini treated with crambene were significantly higher than in untreated acini. Treatment with caspase-3, -8, and -9 inhibitors inhibited annexin V staining, as well as caspase-3 activity, pointing to an important role of these caspases in crambene-induced acinar cell apoptosis. The mitochondrial membrane potential was collapsed, and cytochrome c was released from the mitochondria in crambene-treated acini. Neither TNF-α nor Fas ligand levels were changed in pancreatic acinar cells after crambene treatment. These results provide evidence for the induction of pancreatic acinar cell apoptosis in vitro by crambene and suggest the involvement of mitochondrial pathway in pancreatic acinar cell apoptosis.

scholarly journals MACC1 Suppresses Cell Apoptosis in Hepatocellular Carcinoma by Targeting the HGF/c-MET/AKT Pathway

2015 ◽  
Vol 35 (3) ◽  
pp. 983-996 ◽  
Author(s):  
Yingmin Yao ◽  
Chanwei Dou ◽  
Zhongtang Lu ◽  
Xin Zheng ◽  
Qingguang Liu
Keyword(s):  
Cell Apoptosis ◽  
Target Genes ◽  
Nuclear Translocation ◽  
Annexin V ◽  
Akt Pathway ◽  
Caspase 9 ◽  
Dapi Staining ◽  
In Vivo Experiments

Background & Aims: To investigate the expression and prognostic value of MACC1 in patients with HCC and identify the mechanism by which MACC1 inhibits HCC cell apoptosis. Methods: MACC1 and p-AKT expression was studied using immunohistochemistry of both HCC tissues and adjacent liver tissues. qRT-PCR and western immunoblotting were used to examine the expression of target genes at the mRNA and protein levels, respectively. The MTT assay was used to assess cell viability, and cell apoptosis was determined by DAPI staining, Annexin V/PI staining and Caspase 3/7 assay. Nude mice were used to perform in vivo experiments. Results: The overexpression of MACC1 was found in HCC tissues and was correlated with poor postsurgical prognosis. There was a positive relationship between MACC1 and p-AKT expression in HCC tissues. In vitro experiments showed that MACC1 repressed HCC cell apoptosis and promoted cell growth. Knockdown of c-MET abolished the anti-apoptotic function of MACC1. Next, MACC1 was verified to activate PI3K/AKT signaling by sensitizing HGF/c-MET signaling in HCC. MACC1 overexpression enhanced the HGF-driven phosphorylation of BAD, Caspase 9 and FKHRL1 and inhibited their pro-apoptotic functions in HCC cells. Finally, MACC1 was shown to inhibit cell apoptosis and promote HCC growth in vivo. Conclusions: This investigation revealed that MACC1 overexpression predicted worse prognosis after liver resection, which was attributed to the repression of HCC cell apoptosis via a molecular mechanism in which MACC1 accelerated the activation of the HGF/c-MET/PI3K/AKT pathway and phosphorylated BAD, Caspase 9 and FKHRL1, ultimately preventing their nuclear translocation and their pro-apoptotic function.

BIX-01294-enhanced chemosensitivity in nasopharyngeal carcinoma depends on autophagy-induced pyroptosis

2020 ◽  
Vol 52 (10) ◽  
pp. 1131-1139
Author(s):  
Qian Li ◽  
Min Wang ◽  
Yan Zhang ◽  
Liuqian Wang ◽  
Wei Yu ◽  
...  
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Caspase 3 ◽  
Southern China ◽  
Annexin V ◽  
In Vivo Studies ◽  
Cell Swelling ◽  
Chemotherapeutic Drugs ◽  
Double Positive

Abstract Nasopharyngeal carcinoma (NPC) is a common cancer in southern China and Southeast Asia. Nowadays, radiotherapy is the therapy of choice for NPC patients, and chemotherapy has been found as an alternative treatment for advanced NPC patients. However, finding novel drugs and pharmacologically therapeutic targets for NPC patients is still urgent and beneficial. Our study showed that BIX-01294 (BIX) can induce autophagic vacuoles formation and conversion of LC3B-I to LC3B-II in NPC cells in both dose- and time-dependent manners. Notably, the combination of BIX and chemotherapeutic drugs significantly decreased the cell viability and increased the lactate dehydrogenase release. Meanwhile, BIX plus cis-platinum (Cis) treatment induced pyroptosis in NPC cells as featured by cell swelling and bubble blowing from the plasma membrane, the increased frequency of annexin V and propidium iodide (PI) double-positive cells, as well as the cleavage of gasdermin E (GSDME) and caspase-3. Moreover, the deficiency of GSDME completely shifted pyroptosis to apoptosis. Furthermore, the inhibition of autophagy by chloroquine and the knockout of ATG5 gene significantly blocked the BIX-induced autophagy as well as pyroptosis in both in vitro and in vivo studies. Our data demonstrated that BIX-combined chemotherapeutic drugs could induce the Bax/caspase-3/GSDME-mediated pyroptosis through the activation of autophagy to enhance the chemosensitivity in NPC.

scholarly journals Propofol Alleviates Apoptosis Induced by Chronic High Glucose Exposure via Regulation of HIF-1α in H9c2 Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-13 ◽  
Author(s):  
Jinjun Pu ◽  
Shun Zhu ◽  
Dandan Zhou ◽  
Lidong Zhao ◽  
Ming Yin ◽  
...  
Keyword(s):  
Cell Viability ◽  
Proinflammatory Cytokines ◽  
High Glucose ◽  
Cell Apoptosis ◽  
Caspase 3 ◽  
Mrna Level ◽  
Pcr Analysis ◽  
H9c2 Cell ◽  
Protective Mechanism ◽  
H9c2 Cells

Background. The sedative anesthetic, propofol, is a cardioprotective agent for hyperglycemia-induced myocardial hypertrophy and dysfunction in rats. However, the specific protective mechanism has not been clarified. Methods and Results. In this experiment, we used H9c2 cells subjected to 22 mM glucose lasting for 72 hours as an in vitro model of cardiomyocyte injury by hyperglycemia and investigated the potential mechanism of propofol against hyperglycemic stress in cells. Propofol (5, 10, or 20 μM) was added to the cell cultures before and during the high glucose culture phases. Cell viability and levels of ROS were measured. The levels of proinflammatory cytokines were tested by ELISA. The levels of SIRT3, SOD2, PHD2, HIF-1α, Bcl-2, P53, and cleaved caspase-3 proteins were detected by western blotting. Our data showed that propofol attenuated high glucose-induced cell apoptosis accompanied by a decrease in the level of reactive oxygen species (ROS) and proinflammatory cytokines. Meanwhile, propofol decreased the apoptosis of H9c2 cells via increasing the expression of Bcl-2, SIRT3, SOD2, and PHD2 proteins and decreasing the expression of cleaved caspase-3, P53, and HIF-1α. Real-time PCR analysis showed that propofol did not significantly change the HIF-1α but increase PHD2 at mRNA level. HIF-1α silence significantly decreased apoptosis and inflammation in H9c2 cell during high glucose stress. Pretreatment of IOX2 (the inhibitor of PHD2) inhibited cell viability until the concentration reached 200 μM during high glucose stress. However, 50 μM TYP (the inhibitor of SIRT3) significantly inhibited cell viability during high glucose stress. Delayed IOX2 treatment for 6 hours significantly inhibited cell viability during high glucose stress. Conclusions. Propofol might alleviate cell apoptosis via SIRT3-HIF-1α axis during high glucose stress.

NR4A3 Possesses Tumor Suppressive Properties like NR4A1 By Regulating Pro-Apoptotic Genes in Aggressive Lymphomas and Serves As a Potential Drug Target for Lymphoma Therapy

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 2674-2674 ◽  
Author(s):  
Alexander JA Deutsch ◽  
Beate Rinner ◽  
Kerstin Wenzl ◽  
Katharina Troppan ◽  
Beata Pursche ◽  
...  
Keyword(s):  
Tumor Suppressor ◽  
Cell Line ◽  
Cell Lines ◽  
Caspase 3 ◽  
Annexin V ◽  
Lymphoma Cell ◽  
Aggressive Lymphoma ◽  
Lymphoma Cells ◽  
Apoptotic Genes

Abstract Recently, we described a significant down-regulation of NR4A1 (Nur77) and NR4A3 (Nor-1)-two members of the orphan nuclear receptors acting together as critical tumor suppressor genes in acute myeloid leukemia- in aggressive lymphoma1. NR4A1 over-expression proved its pro-apoptotic function in aggressive lymphoma cells and its lymphoma suppressive properties in vivo was demonstrated in a xenograft mouse model. Since the role of NR4A3 in aggressive lymphomas and the effects of NR4A3 inducing agents on lymphoma cells are unknown, we aimed to investigate NR4A3 function and the effects of a NR4A3 inducing agent in aggressive lymphoma cells. For functional characterization, NR4A3 was over-expressed in a SuDHL4 lymphoma cell line by using an inducible lentiviral construct followed by various apoptotic assays. Induction of NR4A3 expression led to a significantly higher proportion of induced SuDHL4 cells undergoing apoptosis as demonstrated by DNA cleavage, Annexin V staining and increased caspase 3-7 activity suggesting a functional redundancy to NR4A1 in aggressive lymphoma. To test the tumor suppressor function of NR4A3 in vivo, the stably transduced SuDHL4 lymphoma cell line was further investigated in the NOD scid gamma (NSG) mouse model. Induction of NR4A3 in SuDHL4 abrogated tumor growth in the NSG mice, in contrast to vector control- and uninduced SuDHL4 cells, which formed massive lymphoid tumors. Additionally, four aggressive lymphoma cell lines (Karpas-422 and SuDHL4 as GCB- cell line, RI-1 and U2932 as ABC-cell lines) were treated with a NR4A3 inducing agent, named Thapsigargin followed by cell growth (MTS) and apoptotic (Annexin V staining and Caspase 3-7 activity assay) assays. Treatment with Thapsigargin induced NR4A3 expression accompanied with induction of apoptosis of all four lymphoma cell lines detected by increased percentage of Annexin V positive cell and increased caspase 3/7 activity. Inhibition of NR4A3 by siRNA reduced the apoptotic effects of Thapsigargin. To further compare the transcriptional activity as nuclear receptor of NR4A3 to NR4A1, both receptors were separately over-expressed in our four aggressive lymphoma cell lines followed by mRNA expression analysis of intrinsic (Bad, BIK, BID, BMF, Noxa, BAK, Bax, Puma, Bim, Bcl-2, Bcl-X and Mcl-1) and extrinsic (FasL, Fas, Trail, DR4 and DR5) apoptotic genes. mRNA expression analysis of apoptotic genes in aggressive lymphoma cells demonstrated that NR4A1 and NR4A3 over-expression induced Trail, Bim, Puma, BIK, BID and BAK in a similar pattern. Our data suggest that NR4A3 has a pro-apoptotic function in aggressive lymphoma and define that NR4A3 together with the functionally redundant NR4A1 as novel tumor suppressor involved in aggressive lymphoma development. Hence, NR4A3 and its inducing agents are promising novel targets for drug development in lymphoma therapy. 1. Deutsch AJ, Rinner B, Wenzl K, et al. NR4A1-mediated apoptosis suppresses lymphomagenesis and is associated with a favorable cancer specific survival in patients with aggressive B-cell lymphomas. Blood. 2014. Disclosures No relevant conflicts of interest to declare.

scholarly journals Exendin-4 Protects MIN6 Cells from t-BHP-Induced Apoptosis via IRE1-JNK-Caspase-3 Signaling

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Wen-Jia Chen ◽  
Lin-Xi Wang ◽  
Yan-Ping Wang ◽  
Zhou Chen ◽  
Xiao-Ying Liu ◽  
...  
Keyword(s):  
Cell Apoptosis ◽  
Caspase 3 ◽  
Transmembrane Protein ◽  
Annexin V ◽  
Flow Cytometric Assay ◽  
Flow Cytometric ◽  
Tert Butyl Hydroperoxide ◽  
Microscopy Analysis ◽  
Induced Apoptosis ◽  
Fluorescence Microscopy Analysis

Objectives.This study aimed to explore the effect of exendin-4 on t-BHP-induced apoptosis in pancreaticβcells and the mechanism of action.Methods.Murine MIN6 pancreaticβcells were treated with exendin-4 in the presence or absence of tert-butyl hydroperoxide (t-BHP). Cell survival was assessed by MTT staining. The percentage of apoptotic cells was determined by fluorescence microscopy analysis after Hoechst/PI staining and flow cytometric assay after Annexin V-FITC/PI staining. The activity of caspase-3 was determined using a caspase-3 activity kit. Expression of P-IRE1α, IRE1α, C-Jun N-terminal kinase (JNK), P-JNK, C-JUN, and P-C-JUN was detected by western blotting.Results.Exendin-4 was found to inhibit t-BHP-induced apoptosis in pancreaticβ-cells by downregulating caspase-3 activity. Exendin-4 also inhibited the endoplasmic reticulum transmembrane protein IRE1, the apoptosis-related signaling molecule JNK, and c-Jun activation.Conclusions.Our findings suggest that exendin-4 ultimately reduces t-BHP-inducedβ-cell apoptosis. IRE1-JNK-c-Jun signaling is involved in the exendin-4-mediated modulation ofβ-cell apoptosis.

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