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1.
Int J Mol Sci ; 22(2)2021 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-33435582

RESUMO

The aim of the study was to clarify whether orthodontic forces and periodontitis interact with respect to the anti-apoptotic molecules superoxide dismutase 2 (SOD2) and baculoviral IAP repeat-containing protein 3 (BIRC3). SOD2, BIRC3, and the apoptotic markers caspases 3 (CASP3) and 9 (CASP9) were analyzed in gingiva from periodontally healthy and periodontitis subjects by real-time PCR and immunohistochemistry. SOD2 and BIRC3 were also studied in gingiva from rats with experimental periodontitis and/or orthodontic tooth movement. Additionally, SOD2 and BIRC3 levels were examined in human periodontal fibroblasts incubated with Fusobacterium nucleatum and/or subjected to mechanical forces. Gingiva from periodontitis patients showed significantly higher SOD2, BIRC3, CASP3, and CASP9 levels than periodontally healthy gingiva. SOD2 and BIRC3 expressions were also significantly increased in the gingiva from rats with experimental periodontitis, but the upregulation of both molecules was significantly diminished in the concomitant presence of orthodontic tooth movement. In vitro, SOD2 and BIRC3 levels were significantly increased by F. nucleatum, but this stimulatory effect was also significantly inhibited by mechanical forces. Our study suggests that SOD2 and BIRC3 are produced in periodontal infection as a protective mechanism against exaggerated apoptosis. In the concomitant presence of orthodontic forces, this protective anti-apoptotic mechanism may get lost.


Assuntos
Proteína 3 com Repetições IAP de Baculovírus/genética , Regulação da Expressão Gênica , Ligamento Periodontal/metabolismo , Periodonto/metabolismo , Superóxido Dismutase/genética , Animais , Apoptose/genética , Proteína 3 com Repetições IAP de Baculovírus/metabolismo , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Células Cultivadas , Fibroblastos/citologia , Fibroblastos/metabolismo , Fusobacterium nucleatum/fisiologia , Gengiva/citologia , Gengiva/metabolismo , Interações Hospedeiro-Patógeno , Humanos , Ligamento Periodontal/citologia , Ligamento Periodontal/microbiologia , Periodonto/citologia , Periodonto/microbiologia , Ratos , Superóxido Dismutase/metabolismo
2.
Toxicol Mech Methods ; 31(1): 67-72, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32981412

RESUMO

Twenty-five male Wistar rats (140-170 g) were partitioned into 5 groups (n = 5). 2.5 mg/kg, 5 mg/kg, 10 mg/kg and 20 mg/kg of combine Tartrazine and Erythrosine (T+E; 50:50) were administered for 23 days. Serum urea and creatinine, gene expression and profiling of pro-inflammatory cytokine (Tumor Necrosis Factor- α gene), Caspase-9 and Kidney injury molecule-1 (KIM-1) and histomorphological examination of the kidney were investigated. The fold change of relative gene expression of TNF-α gene showed significantly (p < 0.05) up-regulation in all the treated rats except for the 10 mg/kg T+E treated rats when compared to control rats. Casp-9 and KIM-1 genes were significantly (p < 0.05) up-regulated in low dose treatment (2.5 mg/kg T+E and 5 mg/kg T+E) and down-regulated in high dose treatment (10 mg/kg T+E and 20 mg/kg T+E). However, there was significant (p < 0.05) increase in serum urea concentration in the rats treated with 5 mg/kg T+E and 20 mg/kg T+E while the rats treated with 10 mg/kg T+E indicated a significant (p < 0.05) decrease. Conversely, serum creatinine concentration indicated significant (p < 0.05) increase in10mg/kg T+E and 20 mg/kg T+E treated rats versus the control. From the histomorphological examination of the kidney, there was hypertrophy of the glomeruli in relation to the size of Bowman's capsule in the 10 mg/kg T+E and 20 mg/kg T+E treated rats. Kidney function was impaired as evident in up-regulation of TNF-α gene, KIM-1 gene, and serum urea and creatinine concentration with down-regulation of Casp-9 gene. The combined treatment also tampers with the architecture of the kidney.


Assuntos
Lesão Renal Aguda/induzido quimicamente , Caspase 9/metabolismo , Moléculas de Adesão Celular/metabolismo , Corantes/toxicidade , Eritrosina/toxicidade , Rim/efeitos dos fármacos , Tartrazina/toxicidade , Fator de Necrose Tumoral alfa/metabolismo , Lesão Renal Aguda/enzimologia , Lesão Renal Aguda/genética , Lesão Renal Aguda/patologia , Animais , Caspase 9/genética , Moléculas de Adesão Celular/genética , Creatinina/sangue , Relação Dose-Resposta a Droga , Regulação da Expressão Gênica , Rim/enzimologia , Rim/patologia , Masculino , Ratos Wistar , Fator de Necrose Tumoral alfa/genética , Ureia/sangue
3.
BMC Med Genet ; 21(1): 207, 2020 10 19.
Artigo em Inglês | MEDLINE | ID: mdl-33076854

RESUMO

BACKGROUND: Apoptosis is a type of cell death involved in different pathways inherent to the cell and the evasion from this mechanism has been related to cancer, although this process remains not very well comprehended. Gastric cancer (GC) is one of the most incident and aggressive types of cancer worldwide. In this study, we analyzed the distribution of INDEL variants in GC patients (Case) and individuals from the general population (Control) from the Amazon region, in which GC is remarkably frequent. METHODS: A panel of nine INDEL markers in apoptosis-related genes (BCL2 rs11269260, CASP3 rs4647655, CASP8 rs3834129 and rs59308963, CASP9 rs4645982 and rs61079693, FADD rs4197, FAS rs10562972 and TP53 rs17880560) was developed and genotyped by multiplex PCR in both groups. RESULTS: In our analyses, only marker rs4197 (FADD gene) was associated to GC development as follows: INS/DEL genotype of rs4197 increasing in about 2-fold the chances of developing this type of cancer (P = 0.046; OR = 1.940; 95%CI = 1.011-3.725). CONCLUSION: Our results suggest that rs4197 (FADD gene) might play a role in gastric carcinogenesis in the investigated population. More studies are needed to clarify this relation. Here, we highlight the importance of investigating INDEL variants in genes involved in apoptosis.


Assuntos
Apoptose/genética , Predisposição Genética para Doença/genética , Mutação INDEL , Polimorfismo de Nucleotídeo Único , Neoplasias Gástricas/genética , Biomarcadores Tumorais/genética , Caspase 3/genética , Caspase 8/genética , Caspase 9/genética , Proteína de Domínio de Morte Associada a Fas/genética , Genótipo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Neoplasias Gástricas/diagnóstico , Proteína Supressora de Tumor p53/genética , Receptor fas/genética
4.
Chem Biol Interact ; 330: 109236, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32866467

RESUMO

A series of novel pyrrolopyrimidine urea derivatives were synthesized and evaluated for their anticancer activity against colon cancer cell lines. Compounds showed the remarkable cytotoxic activity on HCT-116 wt cell line. The most potent compound 4c (IC50 = 0.14 µM) induced apoptosis in HCT-116 wt and HCT-116 p53-/- cell lines. Otherwise, treatment of HCT-116 BAX-/-BAK-/- cells with compound 4c didn't lead to activation of apoptosis, suggesting that compound 4c induces apoptotic cell death by activating BAX/BAK-dependent pathway. Moreover, while the compound 4c increase the activation of caspase-3 and caspase-9 levels in HCT-116 wt and HCT-116 p53-/- cells, caspase-3 or caspase-9 activation was not observed in HCT-116 BAX-/-BAK-/- cells. In addition, compound 4c induced mitochondrial apoptosis in cells grown as oncospheroids, which better mimic the in vivo milieu of tumors. 4c treatment also activated JNK along with inhibition of prosurvival kinases such as Akt and ERK 1/2 in HCT-116 wt and HCT-116 p53 -/- cells as well as in HCT-116 BAX-/-BAK-/- cells. Notably, our results indicated that compound 4c induced mitochondrial apoptosis through activation p53-independent apoptotic signaling pathways.


Assuntos
Apoptose/efeitos dos fármacos , Neoplasias do Colo/tratamento farmacológico , Pirimidinas/farmacologia , Pirróis/farmacologia , Proteína Supressora de Tumor p53/fisiologia , Caspase 3/genética , Caspase 9/genética , Linhagem Celular Tumoral , Neoplasias do Colo/patologia , Técnicas de Inativação de Genes , Células HCT116 , Humanos , Mitocôndrias/metabolismo , Pirimidinas/síntese química , Pirimidinas/química , Pirróis/síntese química , Pirróis/química , Proteína Supressora de Tumor p53/genética , Proteína Killer-Antagonista Homóloga a bcl-2/genética , Proteína X Associada a bcl-2/genética
5.
Nat Commun ; 11(1): 3173, 2020 06 23.
Artigo em Inglês | MEDLINE | ID: mdl-32576823

RESUMO

Central nervous system ischemic injury features neuronal dysfunction, inflammation and breakdown of vascular integrity. Here we show that activation of endothelial caspase-9 after hypoxia-ischemia is a critical event in subsequent dysfunction of the blood-retina barrier, using a panel of interrelated ophthalmic in vivo imaging measures in a mouse model of retinal vein occlusion (RVO). Rapid nonapoptotic activation of caspase-9 and its downstream effector caspase-7 in endothelial cells promotes capillary ischemia and retinal neurodegeneration. Topical eye-drop delivery of a highly selective caspase-9 inhibitor provides morphological and functional retinal protection. Inducible endothelial-specific caspase-9 deletion phenocopies this protection, with attenuated retinal edema, reduced inflammation and preserved neuroretinal morphology and function following RVO. These results reveal a non-apoptotic function of endothelial caspase-9 which regulates blood-retina barrier integrity and neuronal survival, and identify caspase-9 as a therapeutic target in neurovascular disease.


Assuntos
Caspase 9/metabolismo , Hipóxia/metabolismo , Isquemia/metabolismo , Oclusão da Veia Retiniana/metabolismo , Lesões do Sistema Vascular/metabolismo , Animais , Barreira Hematorretiniana/metabolismo , Caspase 7/metabolismo , Caspase 9/efeitos dos fármacos , Caspase 9/genética , Morte Celular , Modelos Animais de Doenças , Células Endoteliais/metabolismo , Feminino , Predisposição Genética para Doença/genética , Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Coelhos , Retina/metabolismo , Retina/patologia , Oclusão da Veia Retiniana/tratamento farmacológico , Oclusão da Veia Retiniana/patologia , Lesões do Sistema Vascular/patologia
6.
PLoS One ; 15(4): e0223208, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32302311

RESUMO

The aim of this study was to investigate whether exogenous erythropoietin (EPO) administration attenuates N-methyl-D-aspartate (NMDA)-mediated excitotoxic retinal damage in Wistar rats. The survival rate of retinal ganglion cells (RGCs) were investigated by flat mount analysis and flow cytometry. A total of 125 male Wistar rats were randomly assigned to five groups: negative control, NMDA80 (i.e., 80 nmoles NMDA intravitreally injected), NMDA80 + 10ng EPO, NMDA80 + 50ng EPO, and NMDA80 + 250ng EPO. The NMDA80 + 50ng EPO treatment group was used to evaluate various administrated points (pre-/co-/post- administration of NMDA80). Meanwhile, the transferase dUTP Nick-End Labeling (TUNEL) assay of RGCs, the inner plexiform layer (IPL) thickness and the apoptotic signal transduction pathways of µ-calpain, Bax, and caspase 9 were assessed simultaneously using an immunohistochemical method (IHC). When EPO was co-administered with NMDA80, attenuated cell death occurred through the downregulation of the apoptotic indicators: µ-calpain was activated first (peak at ~18hrs), followed by Bax and caspase 9 (peak at ~40hrs). Furthermore, the images of retinal cross sections have clearly demonstrated that thickness of the inner plexiform layer (IPL) was significantly recovered at 40 hours after receiving intravitreal injection with NMDA80 and 50ng EPO. Exogenous EPO may protect RGCs and bipolar cell axon terminals in IPL by downregulating apoptotic factors to attenuate NMDA-mediated excitotoxic retinal damage.


Assuntos
Apoptose , Eritropoetina/farmacologia , N-Metilaspartato/farmacologia , Fármacos Neuroprotetores/farmacologia , Células Ganglionares da Retina/efeitos dos fármacos , Animais , Caspase 9/genética , Caspase 9/metabolismo , Regulação para Baixo , Masculino , Ratos , Ratos Wistar , Receptores de N-Metil-D-Aspartato/agonistas , Receptores de N-Metil-D-Aspartato/metabolismo , Células Ganglionares da Retina/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
7.
Cell Physiol Biochem ; 54(3): 354-370, 2020 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-32298553

RESUMO

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) - RNA-guided Cas9 endonuclease system has provided a fast and efficient method for precise genome editing in diverse mammalian species, including humans. The CRISPR/Cas9 technology allows generation of modifications into site-specific locations of the selected genes in one major step by carrying deletions, insertions or DNA donor-directed precise sequence modifications. Cas9 forms a nucleoprotein complex with a sequence-specific guide RNA to create double-stranded breaks in complementary DNA target. Further, double-stranded break repair machinery leads to the intended gene modifications. The CRISPR/Cas9 system is widely used technique for genome modification, editing and other biotechnology applications, such as functional annotation, a system for visualization of specific genomic loci and transcriptional control of genes. CRISPR/Cas9-mediated manipulation of the laboratory animal genomes has contributed to the understanding of gene functions and has become a popular approach for modeling human disorders. Furthermore, the growing application of CRISPR-Cas9 system to human genes emerges as an extremely powerful technology for the molecular characterization and treatment of human disease. In this review we present the essential principles of CRISPR/Cas9 technology and the recent advances in its use in translational biomedicine.


Assuntos
Sistemas CRISPR-Cas/genética , Edição de Genes/métodos , Regulação da Expressão Gênica/genética , Terapia Genética/métodos , Animais , Sistemas CRISPR-Cas/fisiologia , Caspase 9/química , Caspase 9/genética , Caspase 9/metabolismo , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas/genética , Quebras de DNA de Cadeia Dupla , Reparo do DNA por Junção de Extremidades/genética , Modelos Animais de Doenças , Engenharia Genética/métodos , Recombinação Homóloga/genética , Humanos , RNA Guia/genética
8.
Oncogene ; 39(22): 4390-4403, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32332923

RESUMO

In epithelial ovarian cancer (EOC), response to platinum (PT)-based chemotherapy dictates subsequent treatments and predicts patients' prognosis. Alternative splicing is often deregulated in human cancers and can be altered by chemotherapy. Whether and how changes in alternative splicing regulation could impact on the response of EOC to PT-based chemotherapy is still not clarified. We identified the splicing factor proline and glutamine rich (SFPQ) as a critical mediator of response to PT in an unbiased functional genomic screening in EOC cells and, using a large cohort of primary and recurrent EOC samples, we observed that it is frequently overexpressed in recurrent PT-treated samples and that its overexpression correlates with PT resistance. At mechanistic level, we show that, under PT treatment, SFPQ, in complex with p54nrb, binds and regulates the activity of the splicing factor SRSF2. SFPQ/p54nrb complex decreases SRSF2 binding to caspase-9 RNA, favoring the expression of its alternative spliced antiapoptotic form. As a consequence, SFPQ/p54nrb protects cells from PT-induced death, eventually contributing to chemoresistance. Overall, our work unveils a previously unreported SFPQ/p54nrb/SRSF2 pathway that in EOC cells plays a central role in regulating alternative splicing and PT-induced apoptosis and that could result in the design of new possible ways of intervention to overcome PT resistance.


Assuntos
Antineoplásicos Alquilantes/farmacologia , Cisplatino/farmacologia , Proteínas de Ligação a DNA/fisiologia , Proteínas de Neoplasias/fisiologia , Neoplasias Ovarianas/tratamento farmacológico , Fator de Processamento Associado a PTB/fisiologia , Proteínas de Ligação a RNA/fisiologia , Fatores de Processamento de Serina-Arginina/fisiologia , Animais , Antineoplásicos Alquilantes/uso terapêutico , Apoptose , Caspase 8/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Inibidores de Caspase/farmacologia , Linhagem Celular Tumoral , Cisplatino/uso terapêutico , Proteínas de Ligação a DNA/antagonistas & inibidores , Proteínas de Ligação a DNA/genética , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Feminino , Técnicas de Silenciamento de Genes , Humanos , Camundongos , Neoplasias Ovarianas/metabolismo , Processamento de RNA , RNA Mensageiro/metabolismo , RNA Neoplásico/metabolismo , Proteínas de Ligação a RNA/antagonistas & inibidores , Proteínas de Ligação a RNA/genética , Recidiva , Spliceossomos/metabolismo
9.
Nat Immunol ; 21(5): 546-554, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32231300

RESUMO

High-dose radiation activates caspases in tumor cells to produce abundant DNA fragments for DNA sensing in antigen-presenting cells, but the intrinsic DNA sensing in tumor cells after radiation is rather limited. Here we demonstrate that irradiated tumor cells hijack caspase 9 signaling to suppress intrinsic DNA sensing. Instead of apoptotic genomic DNA, tumor-derived mitochondrial DNA triggers intrinsic DNA sensing. Specifically, loss of mitochondrial DNA sensing in Casp9-/- tumors abolishes the enhanced therapeutic effect of radiation. We demonstrated that combining emricasan, a pan-caspase inhibitor, with radiation generates synergistic therapeutic effects. Moreover, loss of CASP9 signaling in tumor cells led to adaptive resistance by upregulating programmed death-ligand 1 (PD-L1) and resulted in tumor relapse. Additional anti-PD-L1 blockade can further overcome this acquired immune resistance. Therefore, combining radiation with a caspase inhibitor and anti-PD-L1 can effectively control tumors by sequentially blocking both intrinsic and extrinsic inhibitory signaling.


Assuntos
Antineoplásicos Imunológicos/uso terapêutico , Caspase 9/metabolismo , Inibidores de Caspase/uso terapêutico , Quimiorradioterapia/métodos , Neoplasias Colorretais/terapia , Ácidos Pentanoicos/uso terapêutico , Animais , Antígeno B7-H1/genética , Antígeno B7-H1/metabolismo , Caspase 9/genética , Linhagem Celular Tumoral , Modelos Animais de Doenças , Resistencia a Medicamentos Antineoplásicos/genética , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Transplante de Neoplasias , Transdução de Sinais , Regulação para Cima
10.
Environ Toxicol ; 35(7): 794-803, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32149475

RESUMO

The continued use of pesticides is one of the requirements of modern agriculture. Investigations have shown that pesticides can alter gene methylation and expression and subsequently may lead to abortion or birth of embryos with teratogenic disorders. In present study, 30 female NMRI mouse were divided in three experimental groups which in the CPF group, intraperitoneal chlorpyrifos was injected, in the sham group, DMSO was injected, and the control group without injection. The mice were mated and utinized 10 days' post gestation. The number of embryos in each fertilized female, maternal weight, and liver fibrosis was evaluated. The apoptosis pathway genes (caspase3, caspase9) and protein expressions (pro-caspase3, caspase3) of the embryos were evaluated with qRT-PCR and western blot, respectively. The DNA methylation of caspase3 and caspase9 were also assessed. The number of embryos and obtained maternal weight from the CPF group was significantly lower than other two groups. The mRNA expression of Caspase3 and Caspase9 were significantly higher in the CPF group. The protein expression evaluation confirmed the results achieved at the mRNA level. The percentage of Caspase9 DNA methylation in embryos collected from the CPF group was higher compared to the others. It can be considered that consumption of chlorpyrifos toxin can alter the DNA methylation and increase the expression of apoptotic genes. Therefore, continuous use of chlopyrifos may affect pregnancy by increasing the apoptosis pathway in the developing embryos which may lead to abortion or teratogenic disorders in newborn infants.


Assuntos
Apoptose/genética , Clorpirifos/toxicidade , Metilação de DNA/efeitos dos fármacos , Desenvolvimento Embrionário/genética , Exposição Materna/efeitos adversos , Organogênese/genética , Animais , Apoptose/efeitos dos fármacos , Caspase 3/genética , Caspase 9/genética , Desenvolvimento Embrionário/efeitos dos fármacos , Feminino , Camundongos , Organogênese/efeitos dos fármacos , Praguicidas/toxicidade , Gravidez , RNA Mensageiro/metabolismo , Teratogênios/toxicidade
11.
J Exp Clin Cancer Res ; 39(1): 32, 2020 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-32039741

RESUMO

BACKGROUND: Exosomes are essential for tumor growth, metastasis, and are used as novel signaling molecules in targeted therapies. Therefore, exosomal miRNAs can be used in new diagnostic and therapeutic approaches due to their involvement in the development of cancers. However, the detailed biological function, potential molecular mechanism and clinical application of exo-miR-15b-3p in gastric cancer (GC) remains unclear. METHODS: miR-15b-3p mRNA levels in tissues, serum, cells and exosomes were analyzed using qRT-PCR assays. qRT-PCR, immunohistochemical and western blotting analyses were utilized for the determination of DYNLT1 expression. The interrelationship connecting miR-15b-3p with DYNLT1 was verified using Dual-luciferase report, western blotting and qRT-PCR assays. Fluorescent PKH-26 or GFP-Lv-CD63 labeled exosomes, as well as Cy3-miR-15b-3p, were utilized to determine the efficacy of the transfer of exo-miR-15b-3p between BGC-823 and recipient cells. Several in vitro assays and xenograft tumor models were conducted to determine exo-miR-15b-3p impact on GC progression. RESULTS: This is the first study to confirm high miR-15b-3p expression in GC cell lines, tissues and serum. Exosomes obtained from 108 GC patient serum samples and GC cell-conditioned medium were found to show upregulation of exo-miR-15b-3p, with the area under the ROC curve (AUC) being 0.820 [0.763-0.876], which is superior to the AUC of tissues and serum miR-15b-3p (0.674 [0.600-0.748] and 0.642 [0.499-0.786], respectively). In addition, high exo-miR-15b-3p expression in serum was found to accurately predict worse overall survival. SGC-7901 and GES-1 cells are capable of internalizing BGC-823 cell-derived exosomes, allowing the transfer of miR-15b-3p. Migration, invasion, proliferation and inhibition of apoptosis in vitro and in vivo were enhanced by exo-miR-15b-3p, by restraining DYNLT1, Cleaved Caspase-9 and Caspase-3 expression. CONCLUSIONS: This study identified a previously unknown regulatory pathway, exo-miR-15b-3p/DYNLT1/Caspase-3/Caspase-9, which promotes GC development and GES-1 cell malignant transformation. Therefore, serum exo-miR-15b-3p may be a potential GC diagnosis and prognosis biomarker, which can be used in precise targeted GC therapy.


Assuntos
Caspase 3/metabolismo , Caspase 9/metabolismo , Transformação Celular Neoplásica/genética , Dineínas/metabolismo , Exossomos/metabolismo , MicroRNAs/genética , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Idoso , Animais , Apoptose/genética , Biomarcadores Tumorais , Caspase 9/genética , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Modelos Animais de Doenças , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Genes Reporter , Humanos , Masculino , Camundongos , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Transporte de RNA , Transdução de Sinais , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/mortalidade
12.
Virology ; 540: 160-164, 2020 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-31928997

RESUMO

The cancer-causing Epstein-Barr virus (EBV) activates the transcription factor STAT3 upon infecting B-lymphocytes. STAT3 then activates caspase 7 to degrade cellular claspin, resulting in impaired Chk1 phosphorylation. This blockade of ATR-Chk1 signaling allows EBV-transformed cells to proliferate despite DNA lesions from virus-induced replication stress. In addressing the mechanism of caspase 7 activation, we now report that in newly-infected B-cells, STAT3 transcriptionally activates the initiator caspase, caspase 9. Caspase 9 then activates caspase 7 to impair phosphorylation of Chk1 at S345. Importantly, although cleaved products of caspase 9 are detectable in infected cells, there is simultaneous increase in the alternatively-spliced dominant-negative form of caspase 9 - and - expression of dominant-negative caspase 9 is abrogated when STAT3 activation is impaired. Thus EBV, via STAT3, activates caspase 9 but also shifts the balance of transcripts towards its dominant-negative form to allow activation of caspase 7 while avoiding death of EBV-infected cells.


Assuntos
Apoptose , Linfócitos B/metabolismo , Linfócitos B/virologia , Caspase 9/metabolismo , Transformação Celular Viral , Fator de Transcrição STAT3/metabolismo , Linfócitos B/patologia , Caspase 7/metabolismo , Caspase 9/genética , Herpesvirus Humano 4/fisiologia , Humanos , Modelos Biológicos , Fosforilação , RNA Interferente Pequeno , Fator de Transcrição STAT3/genética
13.
Int J Oncol ; 56(1): 368-378, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31789392

RESUMO

Meridianin C is a marine natural product with anticancer activity. Several meridianin C derivatives (compounds 7a­j) were recently synthesized, and their inhibitory effects on pro­viral integration site for Moloney murine leukemia virus (PIM) kinases, as well as their antiproliferative effects on human leukemia cells, were reported. However, the anti­leukemic effects and mechanisms of action of meridianin C and its derivatives remain largely unknown. The aim of the present study was to investigate the effects of meridianin C and its derivatives on MV4­11 human acute myeloid leukemia cell growth. The parent compound meridianin C did not markedly affect the viability and survival of MV4­11 cells. By contrast, MV4­11 cell viability and survival were reduced by meridianin C derivatives, with compound 7a achieving the most prominent reduction. Compound 7a notably inhibited the expression and activity of PIM kinases, as evidenced by reduced B­cell lymphoma­2 (Bcl­2)­associated death promoter phosphorylation at Ser112. However, meridianin C also suppressed PIM kinase expression and activity, and the pan­PIM kinase inhibitor AZD1208 only slightly suppressed the survival of MV4­11 cells. Thus, the anti­survival effect of compound 7a on MV4­11 cells was unrelated to PIM kinase inhibition. Moreover, compound 7a induced apoptosis, caspase­9 and ­3 activation and poly(ADP­ribose) polymerase (PARP) cleavage, but did not affect death receptor (DR)­4 or DR­5 expression in MV4­11 cells. Compound 7a also induced the generation of cleaved Bcl­2, and the downregulation of myeloid cell leukemia (Mcl)­1 and X­linked inhibitor of apoptosis (XIAP) in MV4­11 cells. Furthermore, compound 7a increased eukaryotic initiation factor (eIF)­2α phosphorylation and decreased S6 phosphorylation, whereas GRP­78 expression was unaffected. Importantly, treatment with a pan­caspase inhibitor (z­VAD­fmk) significantly attenuated compound 7a­induced apoptosis, caspase­9 and ­3 activation, PARP cleavage, generation of cleaved Bcl­2 and downregulation of Mcl­1 and XIAP in MV4­11 cells. Collectively, these findings demonstrated the strong anti­survival and pro­apoptotic effects of compound 7a on MV4­11 cells through regulation of caspase­9 and ­3, Bcl­2, Mcl­1, XIAP, eIF­2α and S6 molecules.


Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Apoptose , Proliferação de Células , Indóis/química , Indóis/farmacologia , Leucemia Mieloide Aguda/tratamento farmacológico , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-pim-1/antagonistas & inibidores , Pirimidinas/química , Pirimidinas/farmacologia , Proteínas Reguladoras de Apoptose/genética , Caspase 9/genética , Caspase 9/metabolismo , Humanos , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/patologia , Fosforilação , Inibidores de Proteínas Quinases/química , Células Tumorais Cultivadas , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/genética , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X/metabolismo
14.
Andrologia ; 52(1): e13453, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31762071

RESUMO

miRNAs (MicroRNAs), known as noncoding and important endogenous factors regulating the expression protein-coding genes, are vital regulators in each biological process. Thus, this study aims to explore the key role of four microRNAs in regulating the spermatogenesis. To conduct this experiment, 55 infertile and fertile men provided the study with the sperm and testicular tissue samples. To study the spermatozoa in terms of the morphology, Diff-Quick was applied. Then, quantitative real-time polymerase chain reaction (RT-PCR) was conducted on samples. Our data indicated that in contrast to the miR-15b, significant increasing of miR-383 and miR-122 occurred in both severe oligoasthenoteratozoospermia (SOAT) and moderate oligoasthenoteratozoospermia (MOAT) compared to normal sperm group (N). In addition, it was observed that miR-15b and miR-122 increased in patients with nonobstructive azoospermia (NOA) compared with obstructive azoospermia (OA) group. Expression levels of target genes including P53, CASPASE-9 and CYCLIN D1 underwent principle changes according to miRNAs expression level. Our finding indicated that miRNAs had essential role in the regulation of spermatogenesis, and their expression altering was associated with sperm abnormalities. Thus, microRNAs can be introduced as useful biomarkers to determine male infertility reasons to choose the effective treatment.


Assuntos
Azoospermia/diagnóstico , Redes Reguladoras de Genes , MicroRNAs/metabolismo , Oligospermia/diagnóstico , Espermatogênese/genética , Adulto , Azoospermia/genética , Biomarcadores/análise , Biomarcadores/metabolismo , Caspase 9/genética , Ciclina D1/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Humanos , Masculino , MicroRNAs/análise , Oligospermia/genética , Espermatozoides/metabolismo , Proteína Supressora de Tumor p53/genética , Adulto Jovem
15.
J Photochem Photobiol B ; 202: 111720, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31841988

RESUMO

It has been widely reported that ultraviolet-B (UV-B) radiation is the main extrinsic etiological agent that causes skin photodamage. UV-B exposure mediated photodamage (photo-aging/photo-carcinogenesis) to human skin is caused due to several physiological events at tissue, cellular and molecular levels that lead to impairment of skin function and integrity. In the present study, we investigated the protective role of Trigonelline (TG) against UV-B induced photo-damage in Human Dermal Fibroblasts (Hs68 cells) and Balb/C mice. We exposed human skin fibroblasts and Balb/C mice to UV-B radiation and evaluated various parameters of cellular damage, including, oxidative stress, cytosolic calcium (Ca2+) levels, apoptotic and ER-stress marker proteins. We found that UV-B irradiation induced ROS generation lead to the depletion of endoplasmic reticulum (ER) calcium and increased the expression of ER stress protein markers (phosphorylated elf2α, CHOP, ATF4) as well as apoptotic protein markers (Bcl2, Bax and caspase-9) in a dose and time dependent manner in Hs68 cells. We then determined the effect of TG treatment on UV-B -induced cell death in Hs68 cells and observed that cells exposed to UV-B radiation and treated with TG had a significantly higher survival rate compared to cells exposed to UV-B radiation alone. TG treatment successfully reduced oxidative stress; restored Ca2+ homeostasis and re-established the ER function and prevented apoptotic cell death process. Our results suggest that TG can be used as a potential therapeutic/cosmeceutic agent in preventing skin photo-damage.


Assuntos
Alcaloides/farmacologia , Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Raios Ultravioleta , Animais , Apoptose/efeitos da radiação , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular , Retículo Endoplasmático/metabolismo , Estresse do Retículo Endoplasmático/efeitos da radiação , Fator de Iniciação 1 em Eucariotos/genética , Fator de Iniciação 1 em Eucariotos/metabolismo , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/efeitos da radiação , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo/efeitos da radiação , Espécies Reativas de Oxigênio/metabolismo , Fator de Transcrição CHOP/genética , Fator de Transcrição CHOP/metabolismo
16.
J Photochem Photobiol B ; 203: 111749, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31884347

RESUMO

Gastric cancer (GC) is mainly widespread gastrointestinal malignancy,which reports for 8% of overallcases in carcinogenesis and 10% of yearly fatality, is 4thprimary cause of cancer associated death global. The plan of the present research was to develop ethanolic extract of Vitex negundo-loaded gold nanoparticles (VN-AuNPs) and to appraise the various characteristic methods likes UV-vis spectroscopy, SAED, FTIR, XRD and HR-TEM. Additionally, the anticancer effect of VN-AuNPs on AGS cells were analysed by cell viability, apoptotic morphological changes by TUNEL, AO/EtBr and Hoechst staining, alterations of mitochondrial membrane potential (MMP) and production reactive oxygen species (ROS). Moreover, the status of apoptosis gene such as caspase-3, Bcl-2, Bcl-XL, Bax and caspase-9 expressions was analysed by using western and RT-PCR techniques. Synthesized AuNPs established by UV absorption peak of the highest at 538 and crystal nature of AuNPs was additionallyverifiedwith SAED and XRD. TEM images were illustrates size and morphological division of NPs. FTIR examinationscompletedalkene, carbodiimide and aliphatic primary amines of biomolecules werepresent in synthesized VN-AuNPs. Additionally, AuNPs were stimulatedapoptosis throughthe cytotoxicity effect,changes of MMP, generation of ROS, nuclear and apoptotic morphological alterationsvia TUNEL, AO/EtBr and Hoechst assay. Furthermore, molecular mechanisms also provoked apoptosis through modulating pro (caspase-3, Bax, Bid, caspase-9) and anti-apoptotic (Bcl-2 and Bcl-XL) mediators by western blotting and gene expression in AGS cells. This production of AuNPs from VN was eco-friendly, large-scaled up and easy.


Assuntos
Apoptose/efeitos dos fármacos , Ouro/química , Nanopartículas Metálicas/toxicidade , Vitex/química , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular Tumoral , Química Verde , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Nanopartículas Metálicas/química , Extratos Vegetais/química , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Vitex/metabolismo
17.
J Reprod Dev ; 66(1): 19-27, 2020 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-31735743

RESUMO

One major side effect of chemotherapy that young women with cancer suffer from is ovarian damage. Therefore, it is necessary to study the pathogenesis of chemotherapeutic drugs in order to develop pharmaceutical agents to preserve fertility. Epirubicin is one of the commonly used chemotherapy drugs for breast cancer patients. This research explored the side effects of epirubicin in mice. We found that epirubicin significantly reduced the body weight, the weight of the ovaries and uteri, and the pups' number, while melatonin, which is extremely resistant to oxidation, significantly reduced these damages. Moreover, co-treatment with melatonin prevented epirubicin-induced decrease in E2 and progesterone, and the loss of follicles. Mechanism study showed that melatonin significantly reduced the levels of proapoptotic genes p53, Caspase3, and Caspase9 while it upregulated antiapoptotic factors Bcl-2 and Bcl2l1, and antioxidant genes superoxide dismutase 1 and catalase compared with the epirubicin group. In addition, melatonin markedly reduced reactive oxygen species (ROS) and the transcription of Caspase12 and Chop, which is vital in endoplasmic reticulum stress (ERS)-mediated apoptosis. These results indicate melatonin protects against epirubicin-induced ovarian damage by reducing ROS-induced ERS. Therefore, melatonin has a therapeutic potential for the protection of ovarian function and preservation of fertility during chemotherapy.


Assuntos
Antibióticos Antineoplásicos/toxicidade , Epirubicina/toxicidade , Melatonina/farmacologia , Ovário/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Peso Corporal/efeitos dos fármacos , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Estradiol/sangue , Feminino , Camundongos , Camundongos Endogâmicos ICR , Ovário/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Progesterona/sangue , Espécies Reativas de Oxigênio/metabolismo , Útero/efeitos dos fármacos , Útero/metabolismo
18.
Theriogenology ; 142: 138-148, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31593881

RESUMO

This study was conducted to investigate the effects of maternal exposure bisphenol A (BPA) on ovaries development of F1 female mice. The BPA exposure model of pregnant mice was prepared by intragastric administration of BPA at the doses of 0, 2.5, 5, 10, 20, 40 mg kg-1 d-1 at gestation day (GD) 0.5-17.5. The ovarian index of the offspring mice was calculated at postnatal day (PND) 21 and PND 56. The results showed that BPA at 5 mg/kg, 10 mg/kg, 20 mg/kg and 40 mg/kg significantly increased the abortion rate of the pregnant mice, and each dose of BPA significantly reduced the survival rate of the pups (P < 0.01 or P < 0.05). Besides, there was a non-monotonic dose-response relationship between serum hormone, ovarian receptor levels and BPA in F1 females at both PND 21 and 56. BPA increased the ovarian/uterine index in F1 females at both PND 21 and 56, increased the mRNA relative transcript levels of ovarian ERα, PgR and DNA methyltransferase (DNMT) in F1 females at PND 21, while decreased at PND 56 (P < 0.01 or P < 0.05). BPA also increased the relative expression of caspase-7, caspase-9, bax, inhibited the relative expression of bcl-2 in F1 females at both PND 21 and 56, and increased the apoptosis rate in the ovaries in F1 mice at PND 56 (P < 0.01). The number of follicles in the ovary was increased in F1 females at PND 21, and the ovaries were significantly atrophied when sexual maturity (PND 56). Our results indicated that BPA could disturb the contents of DNMT and make reproductive injury to the offspring females.


Assuntos
Compostos Benzidrílicos/toxicidade , Estrogênios não Esteroides/toxicidade , Ovário/efeitos dos fármacos , Ovário/embriologia , Fenóis/toxicidade , Animais , Animais Recém-Nascidos , Caspase 7/genética , Caspase 7/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Relação Dose-Resposta a Droga , Feminino , Camundongos , Gravidez , Efeitos Tardios da Exposição Pré-Natal , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Organismos Livres de Patógenos Específicos , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
19.
J Virol ; 94(2)2020 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-31694944

RESUMO

MicroRNAs (miRNAs) are small noncoding RNAs that regulate gene expression posttranscriptionally by silencing or degrading their targets and play important roles in the host response to pathogenic infection. Although infectious bursal disease virus (IBDV)-induced apoptosis in host cells has been established, the underlying molecular mechanism is not completely unraveled. Here, we show that infection of DF-1 cells by IBDV induced gga-miR-16-5p (chicken miR-16-5p) expression via demethylation of the pre-miR-16-2 (gga-miR-16-5p precursor) promoter. We found that ectopic expression of gga-miR-16-5p in DF-1 cells enhanced IBDV-induced apoptosis by directly targeting the cellular antiapoptotic protein B-cell lymphoma 2 (Bcl-2), facilitating IBDV replication in DF-1 cells. In contrast, inhibition of endogenous miR-16-5p markedly suppressed apoptosis associated with enhanced Bcl-2 expression, arresting viral replication in DF-1 cells. Furthermore, infection of DF-1 cells with IBDV reduced Bcl-2 expression, and this reduction could be abolished by inhibition of gga-miR-16-5p expression. Moreover, transfection of DF-1 cells with gga-miR-16-5p mimics enhanced IBDV-induced apoptosis associated with increased cytochrome c release and caspase-9 and -3 activation, and inhibition of caspase-3 decreased IBDV growth in DF-1 cells. Thus, epigenetic upregulation of gga-miR-16-5p expression by IBDV infection enhances IBDV-induced apoptosis by targeting the cellular antiapoptotic protein Bcl-2, facilitating IBDV replication in host cells.IMPORTANCE Infectious bursal disease (IBD) is an acute, highly contagious, and immunosuppressive disease in young chickens, causing severe economic losses to stakeholders across the globe. Although IBD virus (IBDV)-induced apoptosis in the host has been established, the underlying mechanism is not very clear. Here, we show that infection of DF-1 cells by IBDV upregulated gga-miR-16-5p expression via demethylation of the pre-miR-16-2 promoter. Overexpression of gga-miR-16-5p enhanced IBDV-induced apoptosis associated with increased cytochrome c release and caspase-9 and -3 activation. Importantly, we found that IBDV infection induced expression of gga-miR-16-5p that triggered apoptosis by targeting Bcl-2, favoring IBDV replication, while inhibition of gga-miR-16-5p in IBDV-infected cells restored Bcl-2 expression, slowing down viral growth, indicating that IBDV induces apoptosis by epigenetic upregulation of gga-miR-16-5p expression. These findings uncover a novel mechanism employed by IBDV for its own benefit, which may be used as a potential target for intervening IBDV infection.


Assuntos
Apoptose , Epigênese Genética , Vírus da Doença Infecciosa da Bursa/fisiologia , MicroRNAs/metabolismo , Regulação para Cima , Replicação Viral , Animais , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Linhagem Celular , Galinhas , Citocromos c/genética , Citocromos c/metabolismo , MicroRNAs/genética
20.
Fish Shellfish Immunol ; 97: 648-655, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31830572

RESUMO

There is crosstalk between the immune and reproductive systems in which sexual dimorphism is a common pattern in vertebrates. In recent years, epigenetics has emerged as a way to study the molecular mechanisms involved in gonadal development, those responsible for integrating environmental information that contribute to assigning a specific sexual phenotype (either an ovary or a testis). The knowledge of epigenetic mechanisms in certain molecular processes allows the development of epigenetic markers. In fish gonads, the existence of reproduction-immune system interactions is known, although the epigenetic mechanisms involved are far from clear. Here, we used the zebrafish (Danio rerio) as a model to study the DNA methylation patterns in gonads of two well-known innate immune genes: IL1ß and Casp9. DNA methylation levels were studied by a candidate gene approach at single nucleotide resolution and gene expression analyses were also carried out. Results showed that there was clear sexual dimorphism in the DNA methylation levels of the two immune genes studied, being significantly higher in the testes when compared to the ovaries. In summary, and although further research is needed, this paper presents sexual dimorphic methylation patterns of two immune-related genes, thus sex-biased differences in methylation profiles should considered when analyzing immune responses in fish. Data showed here can help to develop epimarkers with forthcoming applications in livestock and fish farming production, for example, in immune fish diseases or sexual control programs as epigenetic molecular tools to predict environmental pressure in the gonads.


Assuntos
Caspase 9/genética , Metilação de DNA , Gônadas/imunologia , Interleucina-1beta/genética , Caracteres Sexuais , Peixe-Zebra/genética , Animais , Caspase 9/imunologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Interleucina-1beta/imunologia , Masculino , Ovário/imunologia , Testículo/imunologia , Peixe-Zebra/imunologia , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/imunologia
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