Deoxynivalenol Induces Caspase-8-Mediated Apoptosis through the Mitochondrial Pathway in Hippocampal Nerve Cells of Piglet

Deoxynivalenol (DON) is a common trichothecene mycotoxin found worldwide. DON has broad toxicity towards animals and humans. However, the mechanism of DON-induced neurotoxicity in vitro has not been fully understood. This study investigated the hypothesis that DON toxicity in neurons occurs via the mitochondrial apoptotic pathway. Using piglet hippocampal nerve cells (PHNCs), we evaluated the effects of different concentrations of DON on typical indicators of apoptosis. The obtained results demonstrated that DON treatment inhibited PHNC proliferation and led to morphological, biochemical, and transcriptional changes consistent with apoptosis, including decreased mitochondrial membrane potential, mitochondrial release of cytochrome C (CYCS) and apoptosis inducing factor (AIF), and increased abundance of active cleaved-caspase-9 and cleaved-caspase-3. Increasing concentrations of DON led to decreased B-cell lymphoma-2 (Bcl-2) expression and increased expression of BCL2-associated X (Bax) and B-cell lymphoma-2 homology 3 interacting domain death agonist (Bid), which in turn increased transcriptional activity of the transcription factors AIF and P53 (a tumor suppressor gene, promotes apoptosis). The addition of a caspase-8 inhibitor abrogated these effects. These results reveal that DON induces apoptosis in PHNCs via the mitochondrial apoptosis pathway, and caspase-8 is shown to play an important role during apoptosis regulation.

Download Full-text

Deoxynivalenol induces caspase-8-mediated apoptosis through the mitochondrial pathway in hippocampal nerve cells of piglet

10.21203/rs.2.16325/v1 ◽

2019 ◽

Author(s):

Jinjie Wu ◽

Xichun Wang ◽

Yunjing Jiang ◽

Lei Zhu ◽

Li Cao ◽

...

Keyword(s):

Mitochondrial Pathway ◽

Nerve Cells ◽

Caspase 8 ◽

Trichothecene Mycotoxin ◽

Apoptotic Pathway ◽

Apoptosis Pathway ◽

Mitochondrial Apoptosis Pathway ◽

Transcriptional Changes ◽

Apoptosis Regulation

Abstract Background: Deoxynivalenol (DON) is a common trichothecene mycotoxin found throughout the world. DON has broad toxicity in animals and humans. Its neurotoxicity in vitro, however, is still unclear. This study was designed to investigate the hypothesis that DON toxicity in neurons occurs via the mitochondrial apoptotic pathway. Results: Using piglet hippocampal nerve cells (PHNCs), we evaluated the effects of varying concentrations of DON on typical indicators of apoptosis. The results obtained demonstrated that DON treatment inhibited PHNC proliferation and led to morphological, biochemical, and transcriptional changes consistent with apoptosis, including decreased mitochondrial membrane potential, mitochondrial release of CYCS and AIF, and increased abundance of active cleaved-caspase-9 and cleaved-caspase-3. Increasing concentrations of DON led to decreased Bcl-2 expression and increased expression of Bax and Bid, which in turn increased transcriptional activity of the transcription factors AIF and P53. Addition of a caspase-8 inhibitor abrogated these effects. Conclusion: These data reveal that DON induces apoptosis in PHNCs via the mitochondrial apoptosis pathway, and that caspase-8 plays an important role during apoptosis regulation.

Download Full-text

Deoxypodophyllotoxin Exerts Anti-Cancer Effects on Colorectal Cancer Cells Through Induction of Apoptosis and Suppression of Tumorigenesis

International Journal of Molecular Sciences ◽

10.3390/ijms20112612 ◽

2019 ◽

Vol 20 (11) ◽

pp. 2612 ◽

Cited By ~ 5

Author(s):

Chathurika D. B. Gamage ◽

So-Yeon Park ◽

Yi Yang ◽

Rui Zhou ◽

İsa Taş ◽

...

Keyword(s):

Colorectal Cancer ◽

B Cell ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Apoptotic Pathway ◽

Anti Cancer ◽

Cervical Tumors ◽

Skin Xenograft

Deoxypodophyllotoxin (DPT) is a cyclolignan compound that exerts anti-cancer effects against various types of cancers. DPT induces apoptosis and inhibits the growth of breast, brain, prostate, gastric, lung, and cervical tumors. In this study, we sought to determine the effect of DPT on cell proliferation, apoptosis, motility, and tumorigenesis of three colorectal cancer (CRC) cell lines: HT29, DLD1, and Caco2. DPT inhibited the proliferation of these cells. Specifically, the compound-induced mitotic arrest in CRC cells by destabilizing microtubules and activating the mitochondrial apoptotic pathway via regulation of B-cell lymphoma 2 (Bcl-2) family proteins (increasing Bcl-2 associated X (BAX) and decreasing B-cell lymphoma-extra-large (Bcl-xL)) ultimately led to caspase-mediated apoptosis. In addition, DPT inhibited tumorigenesis in vitro, and in vivo skin xenograft experiments revealed that DPT significantly decreased tumor size and tumor weight. Taken together, our results suggest DPT to be a potent compound that is suitable for further exploration as a novel chemotherapeutic for human CRC.

Download Full-text

The regulation of miR-320a/XBP1 axis through LINC00963 for endoplasmic reticulum stress and autophagy in diffuse large B-cell lymphoma

10.21203/rs.3.rs-353877/v1 ◽

2021 ◽

Author(s):

Yuying Cui ◽

Hui Xu ◽

Yu Yang ◽

Dongmei Zhao ◽

Yu Wen ◽

...

Keyword(s):

B Cell ◽

Caspase 3 ◽

Cell Lymphoma ◽

Xenograft Model ◽

B Cell Lymphoma ◽

Growth Delay ◽

Apoptosis Pathway ◽

Large B Cell Lymphoma ◽

Large B Cell

Abstract Introduction: Huge amounts of gene-sequencing data have been used to guide fundamental researches. The study combined bioinformatics tools with basic study to analyze the pathological mechanisms of diffuse large B-cell lymphoma. Methods: A LncRNA-miRNA-mRNA ceRNA network of diffuse large B cell lymphoma was constructed by GTEx combined with TCGA database analysis. qPCR was used to detect the expression of LINC00963 and miR-320a in DLBCL cell lines. The proteins levels of UPR sensors, GRP78, p-IRE1α, IRE1α, active ATF6, ATF4 and XBP1, were assessed through Western blot, along with apoptosis markers (Bcl-2, Bax, caspase 3) and autophagy indicators (Beclin1, LC3II, LC3I and p62) after LINC00963 overexpression or miR-320a overexpression in vitro. Additionally, the expression of LC3 was analyzed through immunofluorescence (IF) assay. Results: Evaluation of SUDHL4 cell showed marked up-regulation of key elements of the UPR (GRP78, p-IRE1α, spliced XBP-1(XBP-1(s))), apoptosis (Bax, cleaved caspase 3) and autophagy (Beclin1, LC3II) after LINC00963 overexpression in vitro, whereas miR-320a mimic reversed the effects. Besides, LINC00963 targeted miR-320a while miR-320a bound to the 3’UTR of XBP1. The work also found that LINC00963 overexpression resulted in significant tumor growth delay in a xenograft model of DLBCL. Conclusion: Mechanistically, LINC00963 / miR-320a regulated XBP1-apoptosis pathway and autophagy, making this pathway an attractive therapeutic target for selective targeting. The data presented here are the first to comprehensively survey the mechanism of LINC00963 / miR-320a/XBP1 in DLBCL.

Download Full-text

The regulation of miR-320a/XBP1 axis through LINC00963 for endoplasmic reticulum stress and autophagy in diffuse large B-cell lymphoma

Cancer Cell International ◽

10.1186/s12935-021-01992-y ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Yuying Cui ◽

Hui Xu ◽

Yu Yang ◽

Dongmei Zhao ◽

Yu Wen ◽

...

Keyword(s):

B Cell ◽

Caspase 3 ◽

Therapeutic Potential ◽

Cell Lymphoma ◽

Xenograft Model ◽

B Cell Lymphoma ◽

Apoptosis Pathway ◽

Large B Cell Lymphoma ◽

Large B Cell

Abstract Background This study incorporates fundamental research referring to considerable amounts of gene-sequencing data and bioinformatics tools to analyze the pathological mechanisms of diffuse large B-cell lymphoma (DLBCL). Methods A lncRNA-miRNA-mRNA ceRNA network of DLBCL was constructed through database analysis combining GTEx and TCGA. qPCR was used to detect the expression of LINC00963 and miR-320a in DLBCL cell lines. After LINC00963 or miR-320a overexpression in vitro, western blot was performed to assess the protein levels of UPR sensors (GRP78, p-IRE1, IRE1, active ATF6, ATF4 and XBP1), along with apoptosis markers (Bcl-2, Bax, caspase 3) and autophagy indicators (Beclin1, LC3II, LC3I and p62). Additionally, the expression of LC3 was analyzed through immunofluorescence (IF) assay. Results Following LINC00963 overexpression in vitro, SUDHL4 cell line showed a marked increase in the level of UPR-related GRP78, p-IRE1 and spliced XBP-1/XBP-1(s), apoptosis-related Bax and cleaved caspase 3, as well as autophagy-related Beclin1 and LC3II, whereas miR-320a mimic greatly diminished the effects of LINC00963 overexpression. Moreover, LINC00963 targeted miR-320a while miR-320a bound to the 3’UTR of XBP1. It was also found that LINC00963 overexpression resulted in significantly delayed tumor growth in a xenograft model of DLBCL. Conclusion Mechanistically, LINC00963/miR-320a regulated XBP1-apoptosis pathway and autophagy, implying the therapeutic potential of this pathway for selective targeting. The data presented here illustrated the mechanism of LINC00963/miR-320a/XBP1 in DLBCL for the first time.

Download Full-text

Validation of epigenetic mechanisms regulating gene expression in canine B-cell lymphoma: An in vitro and in vivo approach

PLoS ONE ◽

10.1371/journal.pone.0208709 ◽

2018 ◽

Vol 13 (12) ◽

pp. e0208709 ◽

Cited By ~ 2

Author(s):

Silvia Da Ros ◽

Luca Aresu ◽

Serena Ferraresso ◽

Eleonora Zorzan ◽

Eugenio Gaudio ◽

...

Keyword(s):

Gene Expression ◽

B Cell ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Epigenetic Mechanisms

Download Full-text

Targeted inhibition of PI3Kα/δ is synergistic with BCL-2 blockade in genetically defined subtypes of DLBCL

Blood ◽

10.1182/blood-2018-08-872465 ◽

2019 ◽

Vol 133 (1) ◽

pp. 70-80 ◽

Cited By ~ 30

Author(s):

Kamil Bojarczuk ◽

Kirsty Wienand ◽

Jeremy A. Ryan ◽

Linfeng Chen ◽

Mariana Villalobos-Ortiz ◽

...

Keyword(s):

B Cell ◽

Cell Lymphoma ◽

Single Agent ◽

Xenograft Model ◽

Myeloid Cell ◽

B Cell Lymphoma ◽

Cell Receptor ◽

Synergistic Activity ◽

Genetic Bases

Abstract Inhibition of the B-cell receptor (BCR) signaling pathway is a promising treatment strategy in multiple B-cell malignancies. However, the role of BCR blockade in diffuse large B-cell lymphoma (DLBCL) remains undefined. We recently characterized primary DLBCL subsets with distinct genetic bases for perturbed BCR/phosphoinositide 3-kinase (PI3K) signaling and dysregulated B-cell lymphoma 2 (BCL-2) expression. Herein, we explore the activity of PI3K inhibitors and BCL-2 blockade in a panel of functionally and genetically characterized DLBCL cell line models. A PI3K inhibitor with predominant α/δ activity, copanlisib, exhibited the highest cytotoxicity in all BCR-dependent DLBCLs. The proapoptotic effect of copanlisib was associated with DLBCL subtype-specific dysregulated expression of BCL-2 family members including harakiri (HRK) and its antiapoptotic partner BCL extra large (BCL-xL), BCL2 related protein A1, myeloid cell leukemia 1 (MCL-1), and BCL2 interacting mediator of cell death. Using functional BH3 profiling, we found that the cytotoxic activity of copanlisib was primarily mediated through BCL-xL and MCL-1–dependent mechanisms that might complement BCL-2 blockade. For these reasons, we evaluated single-agent activity of venetoclax in the DLBCLs and identified a subset with limited sensitivity to BCL-2 blockade despite having genetic bases of BCL-2 dysregulation. As these were largely BCR-dependent DLBCLs, we hypothesized that combined inhibition of PI3Kα/δ and BCL-2 would perturb BCR-dependent and BCL-2–mediated survival pathways. Indeed, we observed synergistic activity of copanlisib/venetoclax in BCR-dependent DLBCLs with genetic bases for BCL-2 dysregulation in vitro and confirmed these findings in a xenograft model. These results provide preclinical evidence for the rational combination of PI3Kα/δ and BCL-2 blockade in genetically defined DLBCLs.

Download Full-text

Targeting mitochondrial respiration and the BCL2 family in MYC-associated B-cell lymphoma

10.1101/2020.11.22.390922 ◽

2020 ◽

Author(s):

Giulio Donati ◽

Micol Ravà ◽

Marco Filipuzzi ◽

Paola Nicoli ◽

Laura Cassina ◽

...

Keyword(s):

Oxidative Phosphorylation ◽

B Cell ◽

Complex I ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

High Grade ◽

Apoptosis Pathway ◽

Bcl2 Family ◽

Molecular Features ◽

Intrinsic Apoptosis Pathway

AbstractMultiple molecular features, such as activation of specific oncogenes (e. g. MYC, BCL2) or a variety of gene expression signatures, have been associated with disease course in diffuse large B-cell lymphoma (DLBCL). Understanding the relationships between these features and their possible exploitation toward disease classification and therapy remains a major priority in the field. Here, we report that MYC activity in DLBCL is closely correlated with – and most likely a driver of – gene signatures related to Oxidative Phosphorylation (OxPhos). On this basis, we hypothesized that enzymes involved in Oxidative Phosphorylation, and in particular electron-transport chain (ETC) complexes, might constitute tractable therapeutic targets in MYC-associated lymphoma. Indeed, our data show that MYC sensitizes B-cells to IACS-010759, a selective inhibitor of ETC complex I. Mechanistically, IACS-010759 activates an ATF4-driven Integrated Stress Response (ISR), engaging the intrinsic apoptosis pathway through the transcription factor CHOP. In line with these findings, IACS-010759 shows synergy with the BCL2 inhibitor venetoclax against double-hit lymphoma (DHL), a high-grade form of DLBCL with concurrent activation of MYC and BCL2. Similarly, in BCL2-negative lymphoma cell lines, inhibition of the BCL2-related protein Mcl-1 potentiates killing by IACS-010759. Altogether, ETC complex I inhibition engages the ISR to lower the apoptotic threshold in MYC-driven lymphomas and, in combination with select BCL2-family inhibitors, provides a novel therapeutic principle against this aggressive DLBCL subset.Statement of significanceThis work points to OxPhos as a key MYC-activated process and a tractable therapeutic target toward personalized treatment of high-grade DLBCL, providing strong context-dependent cooperation with BH3-mimetic compounds.

Download Full-text

Cernunnos influences human immunoglobulin class switch recombination and may be associated with B cell lymphomagenesis

Journal of Experimental Medicine ◽

10.1084/jem.20110325 ◽

2012 ◽

Vol 209 (2) ◽

pp. 291-305 ◽

Cited By ~ 35

Author(s):

Likun Du ◽

Roujun Peng ◽

Andrea Björkman ◽

Noel Filipe de Miranda ◽

Cornelia Rosner ◽

...

Keyword(s):

B Cell ◽

Cell Lymphoma ◽

Class Switch Recombination ◽

B Cell Lymphoma ◽

Dominant Negative ◽

Dominant Negative Effect ◽

End Joining ◽

Class Switch

Cernunnos is involved in the nonhomologous end-joining (NHEJ) process during DNA double-strand break (DSB) repair. Here, we studied immunoglobulin (Ig) class switch recombination (CSR), a physiological process which relies on proper repair of the DSBs, in B cells from Cernunnos-deficient patients. The pattern of in vivo generated CSR junctions is altered in these cells, with unusually long microhomologies and a lack of direct end-joining. The CSR junctions from Cernunnos-deficient patients largely resemble those from patients lacking DNA ligase IV, Artemis, or ATM, suggesting that these factors are involved in the same end-joining pathway during CSR. By screening 269 mature B cell lymphoma biopsies, we also identified a somatic missense Cernunnos mutation in a diffuse large B cell lymphoma sample. This mutation has a dominant-negative effect on joining of a subset of DNA ends in an in vitro NHEJ assay. Translocations involving both Ig heavy chain loci and clonal-like, dynamic IgA switching activities were observed in this tumor. Collectively, our results suggest a link between defects in the Cernunnos-dependent NHEJ pathway and aberrant CSR or switch translocations during the development of B cell malignancies.

Download Full-text

Characterization of two new high-grade B-cell lymphoma cell lines with MYC and BCL2 rearrangements that are suitable for in vitro drug sensitivity studies

Leukemia & Lymphoma ◽

10.1080/10428194.2018.1508663 ◽

2018 ◽

Vol 60 (4) ◽

pp. 1043-1052

Author(s):

Marie-Sophie Dheur ◽

Hélène A. Poirel ◽

Geneviève Ameye ◽

Gaëlle Tilman ◽

Pascale Saussoy ◽

...

Keyword(s):

B Cell ◽

Cell Lines ◽

Drug Sensitivity ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Lymphoma Cell ◽

High Grade ◽

Sensitivity Studies

Download Full-text

PIWIs maintain testis apoptosis to remove abnormal germ cells in Eriocheir sinensis

Reproduction ◽

10.1530/rep-21-0157 ◽

2021 ◽

Author(s):

Bang-Hong Wei ◽

Jia-Hao Ni ◽

Tong Yang ◽

Shuang-Li Hao ◽

Wan-Xi Yang

Keyword(s):

B Cell ◽

Germ Cells ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Eriocheir Sinensis ◽

Germline Development ◽

P53 Pathway ◽

Apoptosis Pathway ◽

Encoded Proteins

PIWI proteins play important roles in germline development in the mammals. However, the functions of PIWIs in crustaceans remain unknown. In the present study, we identified three Piwis from the testis of Eriocheir sinensis (E. sinensis). Three Piwi genes encoded proteins with typical features of PIWI subfamilies and were highly expressed in the testis. Three PIWIs could be detected in the cytoplasm of spermatocytes and spermatids, while in spermatozoa, we could only detect PIWI1 and PIWI3 in the nucleus. The knockdown of PIWIs by dsRNA significantly affected the formation of the nuclei in spermatozoa, which resulted in deviant and irregular nuclei. PIWI defects significantly inhibited the apoptosis of abnormal germ cells through the caspase-dependent apoptosis pathway and p53 pathway. Knockdown of PIWIs inhibited the expression of caspase3, 7, 8, and p53 without affecting Bcl2 (B-cell lymphoma gene 2), Bax (B-cell lymphoma-2-associated X) and BaxI (B-cell lymphoma-2-associated X inhibitor), which further significantly increased abnormal spermatozoa in the knockdown-group crabs. These results show a new role of PIWI proteins in crustaceans that is different from that in mammals. In summary, PIWIs play roles in the formation of the germline nucleus and can maintain apoptosis in abnormal germ cells to remove abnormal germ cells in E. sinensis.

Download Full-text