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1.
Gen Comp Endocrinol ; 285: 113289, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31557469

RESUMO

Light intensity plays an important role in the regulation of growth, behavior, reproduction, and welfare of avian species. Light intensity preference behavior has been suggested to be involved in welfare of birds. This study aims to investigate the effects of different light intensity and dual light intensity choice (DLIC) lighting program on plasma corticosterone (CORT), and tryptophan hydroxylase 2 (TPH2, the rate-limiting enzyme of serotonin biosynthesis) and tyrosine hydroxylase (TH, the rate-limiting enzyme of dopamine biosynthesis) gene expression in the brainstem of male chickens. Day old broilers were housed in two commercial houses, and placed in 24 pens. All the treatment groups were provided with 23 h light (L) /1 h dark (D) and 30 lx (lx) light intensity during the first week and then 18L:6D (10 lx) from day 7 to 14. Blood and brain were sampled at 14 days of age (10 lx) before the onset of light treatments. On day 15, four treatments (2, 10, 20, and 100 lx), and DLIC treatment (2/20 lx) were initiated. Samples were collected on days 15, 16, 17, 30 and 41. TPH2 expression in the dorsal raphe nucleus (DRN) and caudal raphe nucleus (CRN) of brainstem, and TPH2 and TH expression in ventral tegmental areas (VTN) of the midbrain were determined by qPCR. Results showed that bright light and DLIC lighting program temporarily attenuated plasma CORT, suggesting the short-term stress attenuating effect of bright light and DLIC lighting program. Differential TPH2 expression in the DRN and CRN observed in the DLIC birds indicate a significant effect of DLIC lighting program on the serotonergic activity in the avian brainstem. At the 41 days of age, the significant downregulation of TPH2 and TH expression occurred in the VTA of DLIC treated birds compared to the other group of birds. Taken together, temporal and spatial regulation of TPH2 and TH expression by DLIC lighting program indicate that compensatory regulation of serotonergic and dopaminergic activities might be involved in the light intensity preference behavior of birds, suggesting a possible beneficial effect of the DLIC lighting program on broiler welfare.


Assuntos
Galinhas/sangue , Galinhas/metabolismo , Corticosterona/sangue , Dopamina/metabolismo , Luz , Serotonina/metabolismo , Animais , Tronco Encefálico/metabolismo , Tronco Encefálico/efeitos da radiação , Galinhas/crescimento & desenvolvimento , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Masculino , Núcleos da Rafe/metabolismo , Núcleos da Rafe/efeitos da radiação , Triptofano Hidroxilase/genética , Triptofano Hidroxilase/metabolismo , Tirosina 3-Mono-Oxigenase/genética , Tirosina 3-Mono-Oxigenase/metabolismo , Área Tegmentar Ventral/metabolismo
2.
Radiat Res ; 192(4): 367-379, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31373871

RESUMO

Radiation-induced pulmonary fibrosis (RIPF) is a chronic, progressive complication of therapeutic irradiation of the thorax. It has been suggested that senescence of type II pneumocytes (AECIIs), an alveolar stem cell, plays a role in the development of RIPF through loss of replicative reserve and via senescent AECII-driven release of proinflammatory and profibrotic cytokines. Within this context, we hypothesized that arachidonate 12-lipoxygenase (12-LOX) is a critical mediator of AECII senescence and RIPF. Treatment of wild-type AECIIs with 12S-hydroxyeicosateraenoic acid (12S-HETE), a downstream product of 12-LOX, was sufficient to induce senescence in a NADPH oxidase 4 (NOX4)-dependent manner. Mice deficient in 12-LOX exhibited reduced AECII senescence, pulmonary collagen accumulation and accumulation of alternatively activated (M2) macrophages after thoracic irradiation (5 × 6 Gy) compared to wild-type mice. Conditioned media from irradiated or 12S-HETE-treated primary pneumocytes contained elevated levels of IL-4 and IL-13 compared to untreated pneumocytes. Primary macrophages treated with conditioned media from irradiated AECII demonstrated preferential M2 type polarization when AECIIs were derived from wild-type mice compared to 12-LOX-deficient mice. Together, these data identified 12-LOX as a critical component of RIPF and a therapeutic target for radiation-induced lung injury.


Assuntos
Células Epiteliais Alveolares/patologia , Araquidonato 12-Lipoxigenase/metabolismo , Senescência Celular/efeitos da radiação , Macrófagos/efeitos da radiação , Pneumonite por Radiação/enzimologia , Células Epiteliais Alveolares/efeitos da radiação , Animais , Araquidonato 12-Lipoxigenase/genética , Feminino , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Interleucina-13/biossíntese , Interleucina-4/biossíntese , Macrófagos/citologia , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Pneumonite por Radiação/genética , Pneumonite por Radiação/imunologia , Pneumonite por Radiação/patologia
3.
J Microbiol Biotechnol ; 29(9): 1453-1459, 2019 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-31387339

RESUMO

Zeaxanthin is an important pigment in the photo-protection mechanism of microalgae. However, zeaxanthin epoxidase, an enzyme involved in the accumulation and conversion of zeaxanthin, has not been extensively studied in microalgae. In this work, we report the expression pattern of zeaxanthin epoxidase in Dunaliella tertiolecta (DtZEP) at different light and diverse salinity conditions. To confirm the responsiveness to light conditions, the ZEP expression pattern was investigated in photoperiodic (16 h of light and 8 h of dark) and continuous (24 h of light and 0 h of dark) light conditions. mRNA expression levels in photoperiodic conditions fluctuated along with the light/dark cycle, whereas those in continuous light remained unchanged. In varying salinity conditions, the highest mRNA and protein levels were detected in cells cultured in 1.5 M NaCl, and ZEP expression levels in cells shifted from 0.6 M NaCl to 1.5 M NaCl increased gradually. These results show that mRNA expression of DtZEP responds rapidly to the light/dark cycle or increased salinity, whereas changes in protein synthesis do not occur within a short period. Taken together, we show that DtZEP gene expression responds rapidly to light irradiation and hyperosmotic stress. In addition, ZEP expression patterns in light or salinity conditions are similar to those of higher plants, even though the habitat of D. tertiolecta is different.


Assuntos
Clorofíceas/enzimologia , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Microalgas/enzimologia , Oxirredutases/genética , Vias Biossintéticas/genética , Carotenoides/metabolismo , Clorofíceas/genética , Clorofíceas/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Luz , Microalgas/genética , Microalgas/metabolismo , Oxirredutases/metabolismo , Fotoperíodo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Salinidade , Cloreto de Sódio/farmacologia
4.
Int J Radiat Biol ; 95(11): 1462-1471, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31290713

RESUMO

Protein kinase CK2 is a ubiquitously expressed kinase in eukaryotes, which is known to phosphorylate many protein substrates. Because CK2 is involved in the regulation of various signaling pathways, we wondered whether CK2 participated in the regulation of ionizing radiation (IR) induced biological process. In this study, we investigated the effect of IR on the subcellular localization and kinase activity in human non-small cell lung cancer (NSCLC) cells. Immunofluorescent results showed that CK2 subunits shuttle into the nucleus mostly beginning 1 h after IR and lasting more than 6 h. We also conducted in vitro kinase assay and observed an increase in CK2 kinase activity at 6 h after IR. Furthermore, an increase in S phase was observed at 6 h after IR. Colony formation assay results demonstrated that CK2 inhibitor CX-4945 significantly enhanced the effect of irradiation in NSCLC cells. These results indicated that CK2 may be implicated in the regulation of IR-induced biological process.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/enzimologia , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Neoplasias Pulmonares/enzimologia , Radiação Ionizante , Caseína Quinase II/metabolismo , Ciclo Celular , Linhagem Celular Tumoral , Núcleo Celular/metabolismo , Sobrevivência Celular , Citoplasma/metabolismo , Citosol/metabolismo , Genótipo , Humanos , Naftiridinas/farmacologia , Fosforilação , Transdução de Sinais
5.
Appl Biochem Biotechnol ; 189(3): 729-744, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31111375

RESUMO

Vina-ginsenoside R7 (R7) has been exhibited to engage in multiple pharmacological activities, such as antioxidant and anti-inflammatory activities. However, no photoaging-related studies have been performed on R7. Research is being conducted with the aim of assessing whether treatment with R7 has a protective effect on UVB-induced photoaging skin. Our results show that UVB exposure directly reduces matrix metalloproteinase (MMP) secretion through R7 by restraining the AP-1/MAPK pathway and blocks extracellular matrix (ECM) expression degradation. In addition, R7 improves the expression of transforming growth factor beta 1 (TGF-ß1), and type I procollagen also facilitates the synthesis of collagen by the TGF-ß/Smad signal transduction pathway. Finally, R7 valid blocks nuclear factor-κB (NF-κB) activation and enhances antioxidative stress capacity through activated nuclear factor (erythroid derived 2)-like 2 (Nrf2). In particular, the application of R7 restrains pro-inflammatory cytokines (TNF-α, IL-6, iNOS), which trigger ECM, degrade enzyme production, and suppress vascular endothelial growth factor (VEGF) secretion. In conclusion, R7 may constitute a promising cosmetic ingredient that can protect against skin photodamage resulting from detrimental UVB irradiation.


Assuntos
Anti-Inflamatórios/farmacologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/efeitos da radiação , Ginsenosídeos/farmacologia , Proteínas Luminescentes/farmacologia , Pele/citologia , Raios Ultravioleta/efeitos adversos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Colágeno Tipo I/metabolismo , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/efeitos da radiação , Fibroblastos/citologia , Fibroblastos/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Humanos , Interleucina-6/metabolismo , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Espaço Intracelular/efeitos da radiação , Metaloproteinase 1 da Matriz/genética , Metaloproteinase 1 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/genética , Metaloproteinase 3 da Matriz/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Proteólise/efeitos dos fármacos , Proteólise/efeitos da radiação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/efeitos da radiação , Pele/efeitos dos fármacos , Pele/efeitos da radiação , Fator de Crescimento Transformador beta1/metabolismo , Fator A de Crescimento do Endotélio Vascular/biossíntese
6.
PLoS One ; 14(4): e0215472, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30990828

RESUMO

Early ripening apples are usually used for fresh marketing because of short storage life, although they are with high acid and low sugar contents. Understanding the malate metabolism in fleshy fruit and underpinning process during ripening is crucial for particular crop improvement where acidity is a concern for direct consumption or further processing. In this research, a traditional Chinese apple cultivar 'Hongyu', which belongs to early ripening apple cultivar, were freshly harvested at commercial maturity stage (120 Days after full bloom) and used for different storage temperature (4°C, 20°C) and UV-C treatment (following storage at 20°C after treatment). Simple sugars (glucose, sucrose, and fructose) and organic acids (malic, and oxalic) were assessed after 14 d of storage. Compared to fruits stored at 20°C, the malate content in fruits stored at 4°C significantly higher, while it was decreased significantly in UV-C treated fruits stored at 20°C after 14 d of storage. The sugar content was almost similar throughout the UV-C-treated fruits and fruits stored at different temperature. The higher ratios of total sugars to total organic acids in UV-C treated fruits after 14 d suggest that UV-C treatment has the potential to improve the taste of early ripening apple cultivars. Considering the significant difference in malate the samples at 14 d of storage were subjected for RNA-seq analysis. Transcriptome analysis revealed that the phenomena underlying this change were governed by metabolism of malate by the regulation of NADP-malic enzyme (NADP-ME) and phosphoenolpyruvate carboxylase kinase (PEPCK) in apple during postharvest storage. This transcriptome profiling results have specified the transcript regulation of malate metabolism and lead to possible taste improvement without affecting the other fruit quality attributes.


Assuntos
Armazenamento de Alimentos , Frutas/crescimento & desenvolvimento , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Malato Desidrogenase/biossíntese , Malatos/metabolismo , Malus/crescimento & desenvolvimento , Proteínas de Plantas/biossíntese , Raios Ultravioleta , Perfilação da Expressão Gênica
7.
Acta Biol Hung ; 69(4): 505-509, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30587017

RESUMO

Although the participation of glutathione transferases (GSTs) in light-dependent pathways and the circadian changes in the whole detoxification system have been studied, there are fewer results regarding the exact daily fluctuation of GSTs. In the present study, it was demonstrated that light up-regulated, while dark period decreased the plant GST activity and the expression of the selected tau group GST genes in tomato. These findings provide additional information on our current knowledge on the circadian rhythm of GSTs in plants and could help in further defining detoxification processes.


Assuntos
Ritmo Circadiano/efeitos da radiação , Glutationa Transferase/metabolismo , Luz , Lycopersicon esculentum/efeitos da radiação , Fotoperíodo , Proteínas de Plantas/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Glutationa Transferase/genética , Lycopersicon esculentum/enzimologia , Lycopersicon esculentum/genética , Proteínas de Plantas/genética , Fatores de Tempo
8.
J Biochem ; 164(6): 415-426, 2018 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-30165670

RESUMO

A small nuclear protein, C1D, has roles in various cellular processes, transcription regulation, genome stability surveillance, DNA repair and RNA processing, all of which are required to maintain the host life cycles. In the previous report, C1D directly interacts with XPB, a component of the nucleotide excision repair complex, and C1D knockdown reduced cell survival of 27-1 cells, CHO derivative cells, after UV irradiation. To find out the role of C1D in UV-damaged cells, we used human cell lines with siRNA or shRNA to knockdown C1D. C1D knockdown reduced cell survival rates of LU99 and 786-O after UV irradiation, although C1D knockdown did not affect the efficiency of the nucleotide excision repair. Immunostaining data support that C1D is not directly involved in the DNA repair process in UV-damaged cells. However, H2O2 treatment reduced cell viability in LU99 and 786-O cells. We also found that C1D knockdown upregulated DDIT3 expression in LU99 cells and downregulated APEX1 in 786-O cells, suggesting that C1D functions as a co-repressor/activator. The data accounts for the reduction of cell survival rates upon UV irradiation.


Assuntos
Proteínas Correpressoras/metabolismo , Reparo do DNA/efeitos da radiação , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/metabolismo , DNA/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Estresse Oxidativo/efeitos da radiação , Fator de Transcrição CHOP/metabolismo , Animais , Biomarcadores/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Proteínas Correpressoras/antagonistas & inibidores , Proteínas Correpressoras/genética , Dano ao DNA , Reparo do DNA/efeitos dos fármacos , DNA de Neoplasias/metabolismo , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/antagonistas & inibidores , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/química , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/genética , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Humanos , Peróxido de Hidrogênio/toxicidade , Oxidantes/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Dímeros de Pirimidina/metabolismo , Interferência de RNA , Lesões Experimentais por Radiação/enzimologia , Lesões Experimentais por Radiação/metabolismo , Lesões Experimentais por Radiação/patologia , Fator de Transcrição CHOP/agonistas , Fator de Transcrição CHOP/antagonistas & inibidores , Fator de Transcrição CHOP/genética
9.
Cancer Metastasis Rev ; 37(2-3): 213-225, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29971572

RESUMO

During radiotherapy, an inflammatory response might be induced by activating various enzymes involved in membrane lipid metabolism. The eicosanoid pathway associated with cytosolic phospholipase A2 (cPLA2), cyclooxygenases (COXs), and lipoxygenases (LOXs) can be induced by radiation, and many lipid metabolites might contribute to cancer-associated inflammation, cell proliferation, and cell survival in cancer. The lipid metabolites are also involved in the establishment of the tumor-associated microenvironment through promotion of angiogenesis and formation of vascular network. These biological activities of lipid metabolites are responsible for malignant progression with the acquisition of radioresistance, leading to unsatisfactory outcome of cancer radiotherapy. Many efforts have been made to identify the mechanisms associated with bioactive lipid metabolites and radiation signaling that lead to radioresistance and to develop potent radiosensitizers to improve therapeutic efficacy. Beneficial outcomes would be achieved by targeting the enzymes, such as cPLA2, COXs, and LOXs, responsible for arachidonic acid metabolism and cancer-associated inflammation during cancer radiotherapy. The current study demonstrated a brief review for the radioresistant effects of bioactive lipid metabolites and their enzymes in cancer and the radiosensitizing effects of inhibitors for the enzymes on cancer therapy.


Assuntos
Ácidos Araquidônicos/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Metabolismo dos Lipídeos/efeitos da radiação , Neoplasias/metabolismo , Neoplasias/radioterapia , Animais , Araquidonato 5-Lipoxigenase/metabolismo , Biomarcadores , Ensaios Clínicos como Assunto , Terapia Combinada , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Suscetibilidade a Doenças , Ativação Enzimática/efeitos da radiação , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Metabolismo dos Lipídeos/genética , Neoplasias/enzimologia , Neoplasias/genética , Fosfolipases A2 Citosólicas/genética , Fosfolipases A2 Citosólicas/metabolismo , Prognóstico , Tolerância a Radiação/efeitos dos fármacos , Tolerância a Radiação/genética , Tolerância a Radiação/efeitos da radiação , Resultado do Tratamento
10.
Mol Biol Rep ; 45(5): 1175-1186, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30032382

RESUMO

Ionizing radiation (IR) causes biological effects either by directly damaging the molecules or by generating free radicals. Antioxidant mechanisms are believed to be involved in neutralising free radicals. Levels of antioxidants therefore assume significance in determining the extent of radiation damage. The fruit fly Drosophila melanogaster (D. melanogaster) exhibits remarkable IR tolerance compared to mammals. Present study addresses the questions (1) Whether levels of antioxidants are high in radio-tolerant fruit fly D. melanogaster compared to mammals? (2) Does the antioxidant activity enhance adequately enough post-irradiation? We analysed enzymatic antioxidant profiles and their fluxes prior to and 60 min post-irradiation (50 Gy). Enzymatic antioxidants were analysed in all the developmental stages of D. melanogaster as the fruit fly shows dramatic changes in radiation resistance during development. Activity of superoxide dismutase (SOD) in Drosophila (pre-irradiation) was comparable to that of mammals. Catalase activity was lower than mammals while glutathione peroxidise (DmGPx) activity was significantly higher. Following irradiation SOD showed changes ranging from 1.40 to 1.62 folds only in larval stages. Catalase activity showed positive change of 1.25 folds only in adults. Activity of DmGPx was largely unaffected. Early pupae showed increased (3.67 fold) glutathione S-transferase activity post-irradiation. Non-enzymatic antioxidants such as total antioxidant capacity showed significant whereas reduced glutathione showed insignificant flux. In conclusion, the levels of enzymatic antioxidants in Drosophila compared to IR sensitive mammals and post-irradiation fluxes in antioxidant enzyme levels appear inadequate to explicate the dramatic radiation resistance observed in Drosophila. The observations are in agreement with the recent findings refuting the role of enzymatic antioxidants in radiation resistance.


Assuntos
Antioxidantes/metabolismo , Catalase/metabolismo , Drosophila melanogaster/crescimento & desenvolvimento , Glutationa Peroxidase/metabolismo , Tolerância a Radiação , Superóxido Dismutase/metabolismo , Animais , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/enzimologia , Drosophila melanogaster/efeitos da radiação , Feminino , Raios gama , Regulação da Expressão Gênica no Desenvolvimento/efeitos da radiação , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Masculino , Mamíferos/metabolismo
11.
PLoS One ; 13(6): e0199665, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29953521

RESUMO

Radioresistance is one of the main determinants of treatment outcome in oral squamous cell carcinoma (OSCC), but its prediction is difficult. Several authors aimed to establish radioresistant OSCC cell lines to identify genes with altered expression in response to radioresistance. The development of OSCC is a multistep carcinogenic process that includes activation of several oncogenes and inactivation of tumour suppressor genes. TGM-3 is a tumour suppressor gene and contributes to carcinogenesis process. The aim of this study was to estimate serum and tissue expression of TGM-3 and its correlation with clinico-pathological factors and overall survival in patients of OSCC undergoing chemo-radiotherapy. Tissue expression was observed in formalin fixed tissue biopsies of 96 cases of OSCC and 32 healthy controls were subjected to immunohistochemistry (IHC) by using antibody against TGM-3 and serum level was estimated by ELISA method. mRNA expression was determined by using Real-Time PCR. Patients were followed for 2 year for chemo radiotherapy response. In OSCC, 76.70% cases and in controls 90.62% were positive for TGM-3 IHC expression. TGM-3 expression was cytoplasmic and nuclear staining expressed in keratinized layer, stratum granulosum and stratum spinosum in controls and tumour cells. Mean serum TGM-3 in pre chemo-radiotherapy OSCC cases were 1304.83±573.55, post chemo-radiotherapy samples were 1530.64±669.33 and controls were 1869.16±1377.36, but difference was significant in pre chemo-radiotherapy samples as compared to controls (p<0.018). This finding was also confirmed by real- time PCR analysis in which down regulation (-7.92 fold change) of TGM-3 in OSCC as compared to controls. TGM-3 expression was significantly associated with response to chemo-radiotherapy treatment (p<0.007) and overall survival (p<0.015). Patents having higher level of TGM-3 expression have good response to chemo-radiotherapy and also have better overall survival. TGM-3 may serve as a candidate biomarker for responsiveness to chemo-radiotherapy treatment in OSCC patients.


Assuntos
Carcinoma de Células Escamosas , Quimiorradioterapia , Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Proteínas de Neoplasias/sangue , Tolerância a Radiação , Transglutaminases/sangue , Adulto , Carcinoma de Células Escamosas/enzimologia , Carcinoma de Células Escamosas/mortalidade , Carcinoma de Células Escamosas/terapia , Intervalo Livre de Doença , Feminino , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/enzimologia , Neoplasias Bucais/mortalidade , Neoplasias Bucais/terapia , Taxa de Sobrevida
12.
Support Care Cancer ; 26(11): 3873-3882, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29754212

RESUMO

PURPOSE: Radiotherapy-induced gut toxicity (RIGT) is associated with significant diarrhoea, pain and rectal bleeding. Matrix metalloproteinases (MMPs) have been reported to be involved in chemotherapy-induced gut toxicity and RIGT following single-dose irradiation in vivo. We therefore proposed MMPs would be involved in the pathobiology of RIGT following fractionated irradiation. METHODS: Dark Agouti rats were treated with fractionated radiation (3 × 2.5 Gy/week for 6 weeks). Rats were killed at 3, 6 and 15 weeks to represent acute and chronic toxicities. Sections of jejunum and colon were immunostained for MMP-1, MMP-2, MMP-9 and MMP-14. Relative mRNA expression in jejunum and colon was quantified by RT-PCR for MMP-1, MMP-2, MMP-9 and MMP-14. Western blotting was also conducted on jejunum and colon tissue collected at week 6 to determine protein levels of pro- and active MMP-2. RESULTS: MMP-2 total protein levels, determined by western blotting, significantly increased in both the jejunum (p = 0.0359) and the colon (p = 0.0134) 6 weeks into the fractionated radiation schedule. MMP-1, MMP-2, and MMP-14 mRNA expression significantly increased in the jejunum. MMP-2 mRNA expression was also significantly increased in the colon. Immunostaining of MMP-2 was observed to be increased in both crypt enterocytes and the lamina propria. CONCLUSIONS: MMP-2 plays a role in the pathobiology of gastrointestinal toxicities following fractionated irradiation. Whilst MMP-1 and MMP-14 mRNA expression was increased, this occurred only in the jejunum, suggesting MMPs are differentially involved in RIGT depending on the intestinal region. Further studies are needed to elucidate the role these mediators play in the development and potentiation of RIGT.


Assuntos
Intestino Grosso/metabolismo , Intestino Grosso/efeitos da radiação , Intestino Delgado/metabolismo , Intestino Delgado/efeitos da radiação , Metaloproteinases da Matriz/genética , Lesões por Radiação/genética , Animais , Fracionamento da Dose de Radiação , Relação Dose-Resposta à Radiação , Feminino , Gastroenteropatias/etiologia , Gastroenteropatias/genética , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Mucosa Intestinal/efeitos da radiação , Intestino Grosso/patologia , Intestino Delgado/patologia , Metaloproteinases da Matriz/metabolismo , Doses de Radiação , Lesões por Radiação/patologia , Ratos , Ratos Transgênicos
13.
Radiat Res ; 190(1): 53-62, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29746213

RESUMO

There is a current interest in the development of biodosimetric methods for rapidly assessing radiation exposure in the wake of a large-scale radiological event. This work was initially focused on determining the exposure dose to an individual using biological indicators. Gene expression signatures show promise for biodosimetric application, but little is known about how these signatures might translate for the assessment of radiological injury in radiosensitive individuals, who comprise a significant proportion of the general population, and who would likely require treatment after exposure to lower doses. Using Parp1-/- mice as a model radiation-sensitive genotype, we have investigated the effect of this DNA repair deficiency on the gene expression response to radiation. Although Parp1 is known to play general roles in regulating transcription, the pattern of gene expression changes observed in Parp1-/- mice 24 h postirradiation to a LD50/30 was remarkably similar to that in wild-type mice after exposure to LD50/30. Similar levels of activation of both the p53 and NFκB radiation response pathways were indicated in both strains. In contrast, exposure of wild-type mice to a sublethal dose that was equal to the Parp1-/- LD50/30 resulted in a lower magnitude gene expression response. Thus, Parp1-/- mice displayed a heightened gene expression response to radiation, which was more similar to the wild-type response to an equitoxic dose than to an equal absorbed dose. Gene expression classifiers trained on the wild-type data correctly identified all wild-type samples as unexposed, exposed to a sublethal dose or exposed to an LD50/30. All unexposed samples from Parp1-/- mice were also correctly classified with the same gene set, and 80% of irradiated Parp1-/- samples were identified as exposed to an LD50/30. The results of this study suggest that, at least for some pathways that may influence radiosensitivity in humans, specific gene expression signatures have the potential to accurately detect the extent of radiological injury, rather than serving only as a surrogate of physical radiation dose.


Assuntos
Raios gama/efeitos adversos , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Poli(ADP-Ribose) Polimerase-1/deficiência , Poli(ADP-Ribose) Polimerase-1/genética , Animais , Relação Dose-Resposta à Radiação , Camundongos
14.
Mutat Res ; 809: 13-19, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29625375

RESUMO

The psychrophilic microalga, Chlamydomonas sp. ICE-L, isolated from floating ice in the Antarctic, one of the most highly UV exposed ecosystems on Earth, displays an efficient DNA photorepair capacity. Here, the first known (6-4) photolyase gene (6-4CiPhr) from C. sp. ICE-L was identified. The 6-4CiPhr encoded 559-amino acid polypeptide with a pI of 8.86, and had a predicted Mw of 64.2 kDa. Real-time PCR was carried out to investigate the response of 6-4CiPhr to UVB exposure. The transcription of 6-4CiPhr was up-regulated continuously within 6 h, achieving a maximum of 62.7-fold at 6 h. Expressing 6-4CiPhr in a photolyase-deficient Escherichia coli strain improved survival rate of the strain. In vitro activity assays of purified protein demonstrated that 6-4CiPhr was a photolyase with 6-4PP repair activity. These findings improve understanding of photoreactivation mechanisms of (6-4) photolyase.


Assuntos
Chlamydomonas , Desoxirribodipirimidina Fotoliase , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Proteínas de Plantas , Transcrição Genética/efeitos da radiação , Raios Ultravioleta , Regulação para Cima/efeitos da radiação , Chlamydomonas/enzimologia , Chlamydomonas/genética , Desoxirribodipirimidina Fotoliase/biossíntese , Desoxirribodipirimidina Fotoliase/química , Desoxirribodipirimidina Fotoliase/genética , Proteínas de Plantas/biossíntese , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética
15.
Radiat Res ; 189(5): 519-528, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29474156

RESUMO

Endothelial nitric oxide synthase (eNOS), a constitutive enzyme expressed in vascular endothelial cells, is the main source of nitric oxide (NO), which plays key roles in diverse biological functions, including regulation of vascular tone. Exposure to radiation has been known to generate nitric oxide from eNOS; however, the precise mechanism of its generation and function is not known. The goal of this study was to determine the involvement of radiation-induced DNA damage response (DDR) on eNOS transcription and its effect on cell survival after irradiation. Irradiated bovine aortic endothelial cells showed increased eNOS transcription and NO generation through upregulation of ataxia-telangiectasia mutated (ATM) kinase. Radiation exposure induced NO inhibited cell death, as well as induced cellular senescence postirradiation. This study established that radiation-induced DDR uses ATM kinase to upregulate eNOS transcription and NO generation, leading to cellular senescence, which may play a critical role in radiation-mediated cardiovascular injury.


Assuntos
Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Células Endoteliais/metabolismo , Células Endoteliais/efeitos da radiação , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Óxido Nítrico Sintase Tipo III/genética , Tolerância a Radiação , Animais , Bovinos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Senescência Celular/efeitos dos fármacos , Senescência Celular/efeitos da radiação , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/metabolismo , Citoesqueleto/efeitos da radiação , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/efeitos da radiação , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Óxido Nítrico/farmacologia , Óxido Nítrico Sintase Tipo III/metabolismo , Tolerância a Radiação/efeitos dos fármacos , Transcrição Genética/efeitos dos fármacos , Transcrição Genética/efeitos da radiação
16.
Biosci Biotechnol Biochem ; 82(2): 320-328, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29307270

RESUMO

Although rice bran consumption is reportedly has numerous beneficial effects on human health, the relationship between rice bran and the prevention of photoaging has not been investigated in detail. We sought to investigate whether consumption of rice bran supplement (RBS) can elicit preventive effects against UVB-induced photoaging in vivo. Dorsal skin sections of hairless mice were exposed to UVB over 16 weeks. RBS consumption suppressed UVB-induced wrinkle formation and inhibited the loss of water content and epidermal thickening in the mouse skin. Western blot and immunohistochemical analyses revealed that repeated exposure to UVB upregulated matrix metalloproteinase-13 (MMP-13) and cyclooxygenase-2 (COX-2) expression, while consumption of RBS suppressed MMP-13 and COX-2 expression, as well as mitogen-activated protein kinase (MAPK) signaling pathways. These findings suggest that RBS could be a potential bioactive ingredient in nutricosmetics to inhibit wrinkle formation and water content loss via the suppression of COX-2 and MMP-13 expression.


Assuntos
Suplementos Nutricionais , Oryza/química , Envelhecimento da Pele/efeitos dos fármacos , Envelhecimento da Pele/efeitos da radiação , Raios Ultravioleta/efeitos adversos , Animais , Ciclo-Oxigenase 2/metabolismo , Epiderme/efeitos dos fármacos , Epiderme/patologia , Epiderme/efeitos da radiação , Feminino , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos da radiação , Metaloproteinase 13 da Matriz/metabolismo , Camundongos , Envelhecimento da Pele/patologia , Água/metabolismo
17.
J Cancer Res Clin Oncol ; 144(3): 469-482, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29305710

RESUMO

PURPOSE: Trimodal therapy is a reasonable bladder-preserving option to radical cystectomy. However, many tumors are radioresistive. In this sense, the identification of new prognostic and predictive biomarkers that allow the selection of patients with better responses to radiation therapy would improve outcomes. With the aim of using ecto-5'-nucleotidase/CD73 as a predictive biomarker, the role of this enzyme in the context of radiotherapy in T24 human bladder cancer cell line was investigated. METHODS: T24 cell line was exposure to a single dose of radiation (4 Gray) and trypan blue assay (pharmacological assays of viability/cumulative population doubling), flow cytometry (cell cycle/cell death/active caspase-3/ecto-5'-nucleotidase/CD73 protein staining), DAPI staining (nuclear morphometric assay), RT-PCR and real-time PCR, malachite green method (ectonucleotidase enzymatic assay), and HPLC (analysis of AMP metabolism) were carried out. T24 cell line in which ecto-5'-nucleotidase/CD73 has been completely silenced (5'KO) was also used. RESULTS: The exposure of T24 cell line to a single dose (4 Gray) of radiation-induced cell death and triggered a transitory increase in ecto-5'-nucleotidase/CD73 expression, increased ectonucleotidase activity, and led to adenosine and inosine accumulation in the extracellular medium. Pharmacological inhibition or knocking out ecto-5'-nucleotidase/CD73 rescued cells' proliferative capacity, reducing their sensitivity to radiation. CONCLUSION: Our findings show that the induction of ecto-5'-nucleotidase/CD73 by radiation contributes to the radiosensitivity of T24 cell line.


Assuntos
5'-Nucleotidase/fisiologia , Tolerância a Radiação/genética , Neoplasias da Bexiga Urinária/patologia , Neoplasias da Bexiga Urinária/radioterapia , 5'-Nucleotidase/genética , 5'-Nucleotidase/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/genética , Proliferação de Células/efeitos da radiação , Proteínas Ligadas por GPI/genética , Proteínas Ligadas por GPI/metabolismo , Proteínas Ligadas por GPI/fisiologia , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Técnicas de Silenciamento de Genes , Humanos , Doses de Radiação , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/metabolismo
18.
Mol Cell ; 68(4): 797-807.e7, 2017 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-29149600

RESUMO

DNA lesions caused by UV damage are thought to be repaired solely by the nucleotide excision repair (NER) pathway in human cells. Patients carrying mutations within genes functioning in this pathway display a range of pathologies, including an increased susceptibility to cancer, premature aging, and neurological defects. There are currently no curative therapies available. Here we performed a high-throughput chemical screen for agents that could alleviate the cellular sensitivity of NER-deficient cells to UV-induced DNA damage. This led to the identification of the clinically approved anti-diabetic drug acetohexamide, which promoted clearance of UV-induced DNA damage without the accumulation of chromosomal aberrations, hence promoting cellular survival. Acetohexamide exerted this protective function by antagonizing expression of the DNA glycosylase, MUTYH. Together, our data reveal the existence of an NER-independent mechanism to remove UV-induced DNA damage and prevent cell death.


Assuntos
Dano ao DNA , DNA Glicosilases/metabolismo , Reparo do DNA/efeitos da radiação , Raios Ultravioleta , Acetoexamida/farmacologia , Linhagem Celular Tumoral , DNA Glicosilases/biossíntese , DNA Glicosilases/genética , Reparo do DNA/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Humanos , Masculino
19.
J Biol Chem ; 292(42): 17461-17472, 2017 10 20.
Artigo em Inglês | MEDLINE | ID: mdl-28900036

RESUMO

Polo-like kinase 1 (Plk1), a serine/threonine protein kinase normally expressed in mitosis, is frequently up-regulated in multiple types of human tumors regardless of the cell cycle stage. However, the causal relationship between Plk1 up-regulation and tumorigenesis is incompletely investigated. To this end, using a conditional expression system, here we generated Plk1 transgenic mouse lines to examine the role of Plk1 in tumorigenesis. Plk1 overexpression in mouse embryonic fibroblasts prepared from the transgenic mice led to aberrant mitosis followed by aneuploidy and apoptosis. Surprisingly, Plk1 overexpression had no apparent phenotypes in the mice. Given that no malignant tumor formation was observed even after a long period of Plk1 overexpression, we reasoned that additional factors are required for tumorigenesis in Plk1-overexpressing mice. Because Plk1 can directly participate in the regulation of the DNA damage response (DDR) pathway, we challenged Plk1-overexpressing mice with ionizing radiation (IR) and found that Plk1-overexpressing mice are much more sensitive to IR than their wild-type littermates. Analysis of tumor development in the Plk1-overexpressing mice indicated a marked decrease in the time required for tumor emergence after IR. At the molecular level, Plk1 overexpression led to reduced phosphorylation of the serine/threonine kinases ATM and Chk2 and of histone H2AX after IR treatment both in vivo and in vitro Furthermore, RNA-Seq analysis suggested that Plk1 elevation decreases the expression of several DDR genes. We conclude that Plk1 overexpression may contribute to tumor formation by both inducing chromosomal instability and suppressing the DDR pathway.


Assuntos
Proteínas de Ciclo Celular/biossíntese , Transformação Celular Neoplásica/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Neoplasias Induzidas por Radiação/enzimologia , Proteínas Serina-Treonina Quinases/biossíntese , Proteínas Proto-Oncogênicas/biossíntese , Radiação Ionizante , Animais , Proteínas Mutadas de Ataxia Telangiectasia/genética , Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Proteínas de Ciclo Celular/genética , Transformação Celular Neoplásica/genética , Quinase do Ponto de Checagem 2/genética , Quinase do Ponto de Checagem 2/metabolismo , Dano ao DNA , Regulação Enzimológica da Expressão Gênica/genética , Regulação Neoplásica da Expressão Gênica/genética , Camundongos , Camundongos Transgênicos , Neoplasias Induzidas por Radiação/genética , Neoplasias Induzidas por Radiação/patologia , Fosforilação/genética , Fosforilação/efeitos da radiação , Proteínas Serina-Treonina Quinases/genética , Proteínas Proto-Oncogênicas/genética
20.
Int J Radiat Biol ; 93(11): 1257-1266, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28880721

RESUMO

PURPOSE: The present study was designed to investigate our hypothesis that NADPH oxidase plays a role in radiation-induced pro-oxidative and pro-inflammatory environments in the brain. MATERIALS AND METHODS: C57BL/6 mice received either fractionated whole brain irradiation or sham-irradiation. The mRNA expression levels of pro-inflammatory mediators, such as TNF-α and MCP-1, were determined by quantitative real-time RT-PCR. The protein expression levels of TNF-α, MCP-1, NOX-2 and Iba1 were detected by immunofluorescence staining. The levels of ROS were visualized by in situ DHE fluorescence staining. RESULTS: A significant up-regulation of mRNA and protein expression levels of TNF-α and MCP-1 was observed in irradiated mouse brains. Additionally, immunofluorescence staining of Iba1 showed a marked increase of microglial activation in mouse brain after irradiation. Moreover, in situ DHE fluorescence staining revealed that fractionated whole brain irradiation significantly increased production of ROS. Furthermore, a significant increase in immunoreactivity of NOX-2 was detected in mouse brain after irradiation. On the contrary, an enhanced ROS generation in mouse brain after irradiation was markedly attenuated in the presence of NOX inhibitors or NOX-2 neutralizing antibody. CONCLUSIONS: These results suggest that NOX-2 may play a role in fractionated whole brain irradiation-induced pro-oxidative and pro-inflammatory pathways in mouse brain.


Assuntos
Encéfalo/metabolismo , Encéfalo/efeitos da radiação , Glicoproteínas de Membrana/metabolismo , NADPH Oxidases/metabolismo , Animais , Encéfalo/citologia , Encéfalo/enzimologia , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Fracionamento da Dose de Radiação , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Inflamação/enzimologia , Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microglia/efeitos da radiação , NADPH Oxidase 2 , Oxirredução/efeitos da radiação , Estresse Oxidativo/efeitos da radiação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Regulação para Cima/efeitos da radiação
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