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1.
Nat Commun ; 11(1): 2798, 2020 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-32493925

RESUMO

Mediator 12 (MED12) and MED13 are components of the Mediator multi-protein complex, that facilitates the initial steps of gene transcription. Here, in an Arabidopsis mutant screen, we identify MED12 and MED13 as positive gene regulators, both of which contribute broadly to morc1 de-repressed gene expression. Both MED12 and MED13 are preferentially required for the expression of genes depleted in active chromatin marks, a chromatin signature shared with morc1 re-activated loci. We further discover that MED12 tends to interact with genes that are responsive to environmental stimuli, including light and radiation. We demonstrate that light-induced transient gene expression depends on MED12, and is accompanied by a concomitant increase in MED12 enrichment during induction. In contrast, the steady-state expression level of these genes show little dependence on MED12, suggesting that MED12 is primarily required to aid the expression of genes in transition from less-active to more active states.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Proteínas Repressoras/metabolismo , Arabidopsis/efeitos da radiação , Proteínas de Arabidopsis/genética , Cromatina/metabolismo , Metilação de DNA/genética , Metilação de DNA/efeitos da radiação , Epigênese Genética/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Genes de Plantas , Genes Supressores , Loci Gênicos , Proteínas de Fluorescência Verde/metabolismo , Luz , Plantas Geneticamente Modificadas , Proteínas Repressoras/genética , Regulação para Cima/genética , Regulação para Cima/efeitos da radiação
2.
PLoS One ; 15(3): e0230680, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32218597

RESUMO

Lentinula edodes is one of the most popular edible mushrooms worldwide and contains important medicinal components such as lentinan, ergosterol, and eritadenine. Mushroom metabolism is regulated by the mycelia and fruit body using light; however, in mushrooms, the underlying molecular mechanisms controlling this process as well as light-induced gene expression remain unclear. Therefore, in this study, we compared morphological changes and gene expression in the fruit bodies of L. edodes cultivated under blue light and continuous darkness. Our results showed that blue light primarily induced pileus growth (diameter and thickness) compared to dark cultivation. Alternatively, stipe length development was promoted by dark cultivation. We also performed RNAseq on L. edodes under the blue light/dark cultivation conditions. A total of 12,051 genes were used for aligning the Illumina raw reads and 762 genes that showed fold change cut-offs of >|2| and significance p-values of <0.05 were selected under blue light condition. Among the genes which showed two-fold changed genes, 221 were upregulated and 541 were downregulated. In order to identify blue light induced candidate genes, differentially expressed genes (DEGs) were selected according to 4-fold changes and validated by RT-PCR. We identified 8 upregulated genes under blue light condition, such as DDR48-heat shock protein, Fasciclin-domain-containing protein and carbohydrate esterase family 4 protein, FAD NAD-binding domain-containing protein that are involved in morphological development of primordium and embryonic muscle development, cell adhesion and affect the structure of cellulosic and non-cellulosic cell walls of fruit body development, and photoreceptor of blue light signaling for fruit body and pigment development, respectively. This study provides valuable insights into the molecular mechanisms underlying the role of blue light in mushroom growth and development and can thus contribute to breeding programs to improve mushroom cultivation.


Assuntos
Regulação para Baixo/efeitos da radiação , Luz , Cogumelos Shiitake/genética , Regulação para Cima/efeitos da radiação , Proteínas Fúngicas/genética , RNA Fúngico/química , RNA Fúngico/isolamento & purificação , RNA Fúngico/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de RNA , Cogumelos Shiitake/metabolismo
3.
Medicine (Baltimore) ; 99(8): e19171, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32080097

RESUMO

Circular RNAs (circRNAs), a widespread type of noncoding RNA, are produced by reverse splicing with a circular loop structure. Circ_VCAN (hsa_circ_0073237) acts as a novel circRNA, although its roles in the progression and radioresistance of glioma remain unknown.Expressions of circ_VCAN and microRNA-1183 (miR-1183) were analyzed by quantitative real-time PCR, and the functions of circ_VCAN and irradiate in glioma cell proliferation, apoptosis, migration, and invasion were assessed using cell counting kit-8, flow cytometry, Wound healing, and Transwell assays. The interaction between circ_VCAN and miR-1183 was validated dual-luciferase reporter assay.Our results revealed that circ_VCAN was significantly upregulated in radioresistant glioma tissues compared with radiosensitive tissues, and that circ_VCAN expression was negatively correlated with miR-1183 expression in glioma tissues. We also determined that circ_VCAN expression was decreased and miR-1183 expression was increased in U87 and U251 cells after irradiation. Both knockdown of circ_VCAN and treatment with miR-1183 mimics inhibited proliferation, migration, and invasion, and accelerated apoptosis of the irradiated U87 and U251 cells. In addition, luciferase reporter assays revealed that circ_VCAN might function as a sponge for miR-1183. Finally, overexpression of circ_VCAN expedited carcinogenesis and reduced glioma radiosensitivity by regulating miR-1183.Circ_VCAN serves as a potential oncogene of glioma by regulating miR-1183, and plays an essential role in the radioresistance of glioma.


Assuntos
Neoplasias Encefálicas/genética , Glioma/genética , MicroRNAs/genética , RNA Circular/genética , Versicanas/genética , Apoptose , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/radioterapia , Carcinogênese/genética , Carcinogênese/efeitos da radiação , Movimento Celular , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica/genética , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Glioma/patologia , Glioma/radioterapia , Humanos , Regulação para Cima/genética , Regulação para Cima/efeitos da radiação
4.
Lasers Med Sci ; 35(3): 741-749, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32095920

RESUMO

This in vitro study evaluated the role of photobiomodulation therapy (PBMT) on viability and migration of human dental pulp stem cells (hDPSCs) and its association to epigenetic mechanisms such as histone acetylation. The hDPSCs were characterized and assigned into control and PBMT groups. For the PBMT, five laser irradiations at 6-h intervals were performed using a continuous-wave InGaAlP diode laser. Viability (MTT), migration (scratch), and histone acetylation H3 (H3K9ac immunofluorescence) were evaluated immediately after the last irradiation. PBMT significantly increased the viability (P = 0.004). Also, PBMT group showed significantly increased migration of cells in the wound compared to the control in 6 h (P = 0.002), 12 h (P = 0.014) and 18 h (P = 0.083) being faster than the control, which only finished the process at 24 h. PBMT induced epigenetic modifications in hDPSC due to increased histone acetylation (P = 0.001). PBMT increased viability and migration of hDPSCs, which are related with the upregulation of histone acetylation and could be considered a promising adjuvant therapy for regenerative endodontic treatment.


Assuntos
Movimento Celular/efeitos da radiação , Polpa Dentária/citologia , Histonas/metabolismo , Terapia com Luz de Baixa Intensidade , Células-Tronco/citologia , Células-Tronco/efeitos da radiação , Regulação para Cima/efeitos da radiação , Acetilação/efeitos da radiação , Sobrevivência Celular/efeitos da radiação , Humanos , Células-Tronco/metabolismo
5.
J Photochem Photobiol B ; 204: 111785, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31954267

RESUMO

Postoperative sensory disturbances of inferior alveolar nerve (IAN) are major challenges in dental procedures. We aimed to investigate the effect of photobiomodulation therapy (PBMT) with 810 nm and 980 nm diode lasers on behavioral and immunological factors in a rat IAN crush model. Seventy-two rats were randomly assigned to the four groups of 810 nm laser (crush injury+810 nm laser; 6 J/cm2, 15 sessions, every 48 h), 980 nm laser (crush injury+980 nm laser; same protocol), control (crush injury without irradiation), and sham surgery (no crush injury and no irradiation). The neurosensory response of IAN was evaluated by Von Frey behavioral test before (baseline) and post-surgery in a period of one month. Changes of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), nuclear factor-kappa B (NF-κB), TNF-α, and IL-1ß, were assessed on days 2 and 30 post injury. Data were analyzed for significant differences by repeated measures and one-way ANOVA (p < .05). One day after surgery, all rats subjected to nerve injury showed significant increase in the withdrawal threshold of von Frey test compared to the baseline (p = .02 for control and p = .03 for laser groups). The threshold gradually returned to the baseline scores in 810 nm, 980 nm, and control groups from days 11, 17, and 29, respectively. There was a significant lower withdrawal threshold in 810 nm and 980 nm laser groups compared to the control group in days 11 to 19 and 9 to 23, respectively. At both time points, the levels of NGF and BDNF were significantly higher in 810 nm laser group compared to the control group. There was a significant difference between laser and control groups regarding NF-κB expression (all p values<.001). TNF-α and IL-1ß were significantly lower in laser groups compared to the control group (all p values < .001). PBMT with 810 and 980 nm diode laser protocol used in this study, promoted the neurosensory recovery of IAN after crush injury in rats. In addition, application of 810 nm diode laser was associated with more improvement in immunological responses compared to that of 980 nm laser.


Assuntos
Lasers Semicondutores , Nervo Mandibular/efeitos da radiação , Animais , Comportamento Animal/efeitos da radiação , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Modelos Animais de Doenças , Regulação para Baixo/efeitos da radiação , Interleucina-1beta/metabolismo , Terapia com Luz de Baixa Intensidade , Masculino , Nervo Mandibular/imunologia , Nervo Mandibular/metabolismo , Traumatismos do Nervo Mandibular/imunologia , Traumatismos do Nervo Mandibular/metabolismo , Traumatismos do Nervo Mandibular/radioterapia , NF-kappa B/metabolismo , Fator de Crescimento Neural/metabolismo , Ratos , Ratos Wistar , Fator de Necrose Tumoral alfa/metabolismo , Regulação para Cima/efeitos da radiação
6.
J Photochem Photobiol B ; 203: 111731, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31935633

RESUMO

Idiopathic pulmonary fibrosis (IPF) is a progressive and chronic inflammatory disease with a poor prognosis and very few available treatment options. Low-level laser therapy (LLLT) has been gaining prominence as a new and effective anti-inflammatory and immunomodulatory agent. Can lung inflammation and the airway remodeling be regulated by LLLT in an experimental model of IPF in C57Bl/6 mice? The present study investigated if laser attenuates cellular migration to the lungs, the airway remodeling as well as pro-fibrotic cytokines secretion from type II pneumocytes and fibroblasts. Mice were irradiated (780 nm and 30 mW) and then euthanized fifteen days after bleomycin-induced lung fibrosis. Lung inflammation and airway remodeling were evaluated through leukocyte counting in bronchoalveolar lavage fluid (BALF) and analysis of collagen in lung, respectively. Inflammatory cells in blood were also measured. For in vitro assays, bleomycin-activated fibroblasts and type II pneumocytes were irradiated with laser. The pro- and anti-inflammatory cytokines level in BALF as well as cells supernatant were measured by ELISA, and the TGFß in lung was evaluated by flow cytometry. Lung histology was used to analyze collagen fibers around the airways. LLLT reduced both migration of inflammatory cells and deposition of collagen fibers in the lungs. In addition, LLLT downregulated pro-inflammatory cytokines and upregulated the IL-10 secretion from fibroblasts and pneumocytes. Laser therapy greatly reduced total lung TGFß. Systemically, LLLT also reduced the inflammatory cells counted in blood. There is no statistical difference in inflammatory parameters studied between mice of the basal group and the laser-treated mice. Data obtained indicate that laser effectively attenuates the lung inflammation, and the airway remodeling in experimental pulmonary fibrosis is driven to restore the balance between the pro- and anti-inflammatory cytokines in lung and inhibit the pro-fibrotic cytokines secretion from fibroblasts.


Assuntos
Remodelação das Vias Aéreas , Citocinas/metabolismo , Fibrose Pulmonar Idiopática/radioterapia , Lasers , Animais , Líquido da Lavagem Broncoalveolar/química , Células Cultivadas , Citocinas/análise , Modelos Animais de Doenças , Regulação para Baixo/efeitos da radiação , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Células Epiteliais/efeitos da radiação , Fibroblastos/citologia , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Fibrose Pulmonar Idiopática/patologia , Terapia a Laser , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Regulação para Cima/efeitos da radiação
7.
Biochim Biophys Acta Bioenerg ; 1861(4): 148014, 2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30880080

RESUMO

The spring ephemeral Berteroa incana is a familial relative of Arabidopsis thaliana and thrives in a diverse range of terrestrial ecosystems. Within this study, the novel chlorophyll fluorescence parameter of photochemical quenching in the dark (qPd) was used to measure the redox state of the primary quinone electron acceptor (QA) in order to estimate the openness of photosystem II (PSII) reaction centres (RC). From this, the early onset of photoinactivation can be sensitively quantified alongside the light tolerance of PSII and the photoprotective efficiency of nonphotochemical quenching (NPQ). This study shows that, with regards to A. thaliana, NPQ is enhanced in B. incana in both low-light (LL) and high-light (HL) acclimation states. Moreover, light tolerance is increased by up to 500%, the rate of photoinactivation is heavily diminished, and the ability to recover from light stress is enhanced in B. incana, relative to A. thaliana. This is due to faster synthesis of zeaxanthin and a larger xanthophyll cycle (XC) pool available for deepoxidation. Moreover, preferential energy transfer via CP47 around the RC further enhances efficient photoprotection. As a result, a high functional cross-section of photosystem II is maintained and is not downregulated when B. incana is acclimated to HL. A greater capacity for protective NPQ allows B. incana to maintain an enhanced light-harvesting capability when acclimated to a range of light conditions. This enhancement of flexible short-term protection saves the metabolic cost of long-term acclimatory changes.


Assuntos
Aclimatação/fisiologia , Brassicaceae/fisiologia , Processos Fotoquímicos , Estações do Ano , Aclimatação/efeitos da radiação , Arabidopsis/fisiologia , Arabidopsis/efeitos da radiação , Brassicaceae/efeitos da radiação , Transferência de Energia , Cinética , Luz , Complexo de Proteína do Fotossistema II/metabolismo , Espectrometria de Fluorescência , Estresse Fisiológico/efeitos da radiação , Regulação para Cima/efeitos da radiação , Xantofilas/metabolismo , Zeaxantinas/biossíntese
8.
Int J Mol Sci ; 20(19)2019 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-31547113

RESUMO

Sphingosine 1 phosphate (S1P) is a bioactive lipid that regulates cellular activity, including proliferation, cytoskeletal organization, migration, and fibrosis. In this study, the potential relevance of S1P-Rho signaling in pterygium formation and the effects of ultraviolet (UV) irradiation on activation of the S1P/S1P receptor axis and fibrotic responses were investigated in vitro. Expressions of the S1P2, S1P4, and S1P5 receptors were significantly higher in pterygium tissue than in normal conjunctiva, and the concentration of S1P was significantly elevated in the lysate of normal conjunctival fibroblast cell (NCFC) irradiated with UV (UV-NCFCs). RhoA activity was significantly upregulated in pterygium fibroblast cells (PFCs) and UV-NCFCs, and myosin phosphatase-Rho interacting protein (MRIP) was upregulated, and myosin phosphatase target subunit 1 (MYPT1) was downregulated in PFCs. Fibrogenic changes were significantly upregulated in both PFCs and UV-NCFCs compared to NCFCs. We found that the activation of the S1P receptor-Rho cascade was observed in pterygium tissue. Additionally, in vitro examination showed S1P-rho activation and fibrogenic changes in PFCs and UV-NCFCs. S1P elevation and the resulting upregulation of the downstream Rho signaling pathway may be important in pterygium formation; this pathway offers a potential therapeutic target for suppressing pterygium generation.


Assuntos
Túnica Conjuntiva/metabolismo , Lisofosfolipídeos/metabolismo , Pterígio/metabolismo , Transdução de Sinais/efeitos da radiação , Esfingosina/análogos & derivados , Raios Ultravioleta , Proteínas rho de Ligação ao GTP/metabolismo , Túnica Conjuntiva/patologia , Proteínas do Olho/biossíntese , Feminino , Humanos , Masculino , Pterígio/patologia , Esfingosina/metabolismo , Regulação para Cima/efeitos da radiação
9.
Can J Microbiol ; 65(12): 922-929, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31525298

RESUMO

Blue light (BL) exerts an antimicrobial effect on pathogenic bacteria. It has been hypothesized that its bactericidal activity depends upon the generation of reactive oxygen species (such as anion superoxides) and the resultant cellular damage. However, some aspects of this hypothesis needed to be tested and investigated. Thus, the work conducted herein examined the molecular impact of BL treatment on Cronobacter sakazakii, an emerging foodborne pathogen. The results showed that BL exhibited an efficient bactericidal effect against C. sakazakii. Under a sublethal BL dose, both intracellular anion superoxides and malondialdehyde (a marker of oxidative stress) contents were increased gradually. Moreover, permeability of the outer membrane was increased by approximately 50%, indicating membrane damage. Further investigation revealed alterations to cellular fatty acid profiles, with a decrease and disappearance of unsaturated fatty acids, including C18:2, C16:1, and C18:1. These data indicate that bacterial lipids, especially unsaturated fatty acids, are important molecular targets of BL photo-oxidation. The transcriptional response of bacteria to BL was also studied, and it was found that three genes were upregulated, including genes encoding antioxidants. The current study contributes towards an improved understanding of the bactericidal mechanisms of BL and highlights the importance of lipid and membrane damage.


Assuntos
Cronobacter sakazakii/efeitos da radiação , Ácidos Graxos/efeitos da radiação , Luz , Estresse Oxidativo/efeitos da radiação , Membrana Externa Bacteriana/metabolismo , Membrana Externa Bacteriana/efeitos da radiação , Cronobacter sakazakii/genética , Cronobacter sakazakii/metabolismo , Ácidos Graxos/química , Genes Bacterianos/genética , Viabilidade Microbiana/efeitos da radiação , Espécies Reativas de Oxigênio/metabolismo , Regulação para Cima/efeitos da radiação
10.
Int J Radiat Oncol Biol Phys ; 105(3): 525-536, 2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-31271826

RESUMO

PURPOSE: Proton radiation is a major component of the radiation field in outer space and is used clinically in radiation therapy of resistant cancers. Although epidemiologic studies in atom bomb survivors and radiologic workers have established radiation as a risk factor for colorectal cancer (CRC), we have yet to determine the risk of CRC posed by proton radiation owing to a lack of sufficient human or animal data. The purpose of the current study was to quantitatively and qualitatively characterize differential effects of acute and fractionated high-energy protons on colorectal carcinogenesis. METHODS AND MATERIALS: We used ApcMin/+ mice, a well-studied CRC model, to examine acute versus fractionated proton radiation-induced differences in intestinal tumorigenesis and associated signaling pathways. Mice were exposed to 1.88 Gy of proton radiation delivered in a single fraction or in 4 equal daily fractions (0.47 Gy × 4). Intestinal tumor number and grade were scored 100 to 110 days after irradiation, and tumor and tumor-adjacent normal tissues were harvested to assess proliferative ß-catenin/Akt pathways and DNA damage response and repair pathways relevant to colorectal carcinogenesis. RESULTS: Significantly higher intestinal tumor number and grade, along with decreased differentiation, were observed after acute radiation relative to fractionated radiation. Acute protons induced upregulation of ß-catenin and Akt pathways with increased proliferative marker phospho-histone H3. Increased DNA damage along with decreased DNA repair factors involved in mismatch repair and nonhomologous end joining were also observed after exposure to acute protons. CONCLUSIONS: We show increased γH2AX, 53BP1, and 8-oxo-dG, suggesting that increased ongoing DNA damage along with decreased DNA repair factors and increased proliferative responses could be triggering a higher number of intestinal tumors after acute relative to fractionated proton exposures in ApcMin/+ mice. Taken together, our data suggest greater carcinogenic potential of acute relative to fractionated proton radiation.


Assuntos
Quebras de DNA de Cadeia Dupla , Reparo de Erro de Pareamento de DNA , Neoplasias Intestinais/genética , Neoplasias Induzidas por Radiação/genética , Prótons/efeitos adversos , 8-Hidroxi-2'-Desoxiguanosina/metabolismo , Animais , Carcinogênese/genética , Diferenciação Celular , Proliferação de Células , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Ciclina D1/metabolismo , Reparo do DNA por Junção de Extremidades , Modelos Animais de Doenças , Fracionamento da Dose de Radiação , Feminino , Expressão Gênica , Genes APC , Histonas/metabolismo , Immunoblotting/métodos , Neoplasias Intestinais/patologia , Intestino Delgado/metabolismo , Intestino Delgado/efeitos da radiação , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias Induzidas por Radiação/patologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Doses de Radiação , Exposição à Radiação/efeitos adversos , Voo Espacial , Proteína 1 de Ligação à Proteína Supressora de Tumor p53/metabolismo , Regulação para Cima/efeitos da radiação , beta Catenina/metabolismo
11.
Medicina (Kaunas) ; 55(7)2019 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-31252673

RESUMO

Background: Radiation-induced heart injury can lead to increased risk of heart failure, attack, and ischemia. Some studies proposed IL-4 and IL-13 as two important cytokines that are involved in late effects of ionizing radiation. On the other hand, these cytokines may, through upregulation of Duox1 and Duox2, induce chronic oxidative stress, inflammation, and fibrosis. In this study, we evaluated the upregulation of Duox1 and Duox2 pathways in hearts following chest irradiation in rats and then detected possible attenuation of them by melatonin. Materials and Methods: Twenty male Wistar rats were divided into four groups: (1) control; (2) melatonin treated (100 mg/kg); (3) radiation (15 Gy gamma rays); (4) melatonin treated before irradiation. All rats were sacrificed after 10 weeks and their heart tissues collected for real-time PCR (RT-PCR), ELISA detection of IL-4 and IL-13, as well as histopathological evaluation of macrophages and lymphocytes infiltration. Results: Results showed an upregulation of IL-4, IL4ra1, Duox1, and Duox2. The biggest changes were for IL4ra1 and Duox1. Treatment with melatonin before irradiation could attenuate the upregulation of all genes. Melatonin also caused a reduction in IL-4 as well as reverse infiltration of inflammatory cells. Conclusion: Duox1 and Duox2 may be involved in the late effects of radiation-induced heart injury. Also, via attenuation of these genes, melatonin can offer protection against the toxic effects of radiation on the heart.


Assuntos
Oxidases Duais/efeitos da radiação , Melatonina/farmacologia , Regulação para Cima/efeitos da radiação , Análise de Variância , Animais , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática/métodos , Cardiopatias Congênitas , Masculino , Melatonina/uso terapêutico , Fatores de Proteção , Lesões por Radiação , Ratos , Ratos Wistar , Regulação para Cima/fisiologia
12.
Photodermatol Photoimmunol Photomed ; 35(5): 360-368, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31166622

RESUMO

BACKGROUND/PURPOSE: Ultraviolet (UV) A (315-400 nm) is the UV light that most frequently reaches the Earth's surface and can penetrate the epidermis through to the dermis, causing various issues, including skin aging and skin cancer. The results of our previous studies have shown that the flavonoid monomer cyanidin-3-o-glucoside (C3G) can effectively inhibit primary human dermal fibroblast (HDF) oxidative damage and apoptosis caused by UVA radiation. Many flavonoids can regulate the level of autophagy. However, whether C3G inhibits UVA-induced oxidative damage to primary HDFs by regulating autophagy levels remains unclear. METHODS AND RESULTS: In this study, we used different doses (0-12 J/cm2 ) of UVA to irradiate cells and showed that the expression levels of autophagy-related gene 5 (Atg5) and microtubule-associated protein 1 light chain 3 (LC3)-II in primary HDFs first increased and then decreased. The expression of Atg5 and LC3-II was significantly decreased under 12 J/cm2 (light-damage model). C3G increased the levels of Atg5 and LC3-II. Primary HDFs were pretreated with C3G, followed by treatment with the autophagy inhibitor 3-methyladenine (3-MA) after 12 J/cm2 UVA irradiation. The inhibitory effects of C3G on morphological changes, oxidative damage, and apoptosis in primary HDFs induced by UVA were significantly decreased. CONCLUSION: C3G can inhibit UVA-induced damage to primary HDFs by inducing autophagy. These results provide a theoretical basis for the application of natural compounds to resist light damage to the skin in the future.


Assuntos
Antocianinas/farmacologia , Autofagia , Derme/metabolismo , Fibroblastos/metabolismo , Glucosídeos/farmacologia , Raios Ultravioleta/efeitos adversos , Regulação para Cima , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Autofagia/efeitos dos fármacos , Autofagia/efeitos da radiação , Células Cultivadas , Derme/patologia , Fibroblastos/patologia , Humanos , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/efeitos da radiação
13.
Molecules ; 24(9)2019 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-31060229

RESUMO

Background: KDM5 enzymes are H3K4 specific histone demethylases involved in transcriptional regulation and DNA repair. These proteins are overexpressed in different kinds of cancer, including breast, prostate and bladder carcinomas, with positive effects on cancer proliferation and chemoresistance. For these reasons, these enzymes are potential therapeutic targets. Methods: In the present study, we analyzed the effects of three different inhibitors of KDM5 enzymes in MCF-7 breast cancer cells over-expressing one of them, namely KDM5B/JARID1B. In particular we tested H3K4 demethylation (western blot); radio-sensitivity (cytoxicity and clonogenic assays) and damage accumulation (COMET assay and kinetics of H2AX phosphorylation). Results: we show that all three compounds with completely different chemical structures can selectively inhibit KDM5 enzymes and are capable of increasing sensitivity of breast cancer cells to ionizing radiation and radiation-induced damage. Conclusions: These findings confirm the involvement of H3K4 specific demethylases in the response to DNA damage, show a requirement of the catalytic function and suggest new strategies for the therapeutic use of their inhibitors.


Assuntos
Neoplasias da Mama/enzimologia , Histona Desmetilases/antagonistas & inibidores , Histona Desmetilases com o Domínio Jumonji/genética , Proteínas Nucleares/genética , Radiossensibilizantes/farmacologia , Proteínas Repressoras/genética , Bibliotecas de Moléculas Pequenas/farmacologia , Neoplasias da Mama/genética , Neoplasias da Mama/terapia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Histonas/metabolismo , Humanos , Histona Desmetilases com o Domínio Jumonji/metabolismo , Células MCF-7 , Modelos Moleculares , Estrutura Molecular , Proteínas Nucleares/metabolismo , Tolerância a Radiação/efeitos dos fármacos , Radiossensibilizantes/química , Proteínas Repressoras/metabolismo , Bibliotecas de Moléculas Pequenas/química , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/efeitos da radiação
14.
Int J Mol Sci ; 20(10)2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-31096691

RESUMO

Blue light (BL) plays an important role in regulation of the growth and development of aquatic plants and land plants. Aureochrome (AUREO), the recent BL photoreceptor identified in photosynthetic stramenopile algae, is involved in the photomorphogenesis and early development of Saccharina japonica porophytes (kelp). However the factors that interact with the SjAUREO under BL conditions specifically are not clear. Here in our study, three high quality cDNA libraries with CFU over 5 × 106 and a recombination rate of 100% were constructed respectively through white light (WL), BL and darkness (DK) treatments to the juvenile sporophytes. Based on the constructed cDNA libraries, the interactors of SjAUREO were screened and analyzed. There are eighty-four genes encoding the sixteen predicted proteins from the BL cDNA library, sixty-eight genes encoding eighteen predicted proteins from the DK cDNA library, and seventy-four genes encoding nineteen proteins from the WL cDNA library. All the predicted proteins are presumed to interact with SjAUREO when co-expressed with SjAUREO seperately. The 40S ribosomal protein S6 (RPS6), which only exists in the BL treated cDNA library except for two other libraries, and which is essential for cell proliferation and is involved in cell cycle progression, was selected for detailed analysis. We showed that its transcription was up-regulated by BL, and was highly transcribed in the basal blade (meristem region) of juvenile sporophytes but less in the distal part. Taken together, our results indicated that RPS6 was highly involved in BL-mediated kelp cellular division and photomorphogenesis by interacting with SjAUREO.


Assuntos
Laminaria/metabolismo , Laminaria/efeitos da radiação , Luz , Proteína S6 Ribossômica/metabolismo , Proteína S6 Ribossômica/efeitos da radiação , Proteínas Ribossômicas/metabolismo , Proteínas Ribossômicas/efeitos da radiação , Proliferação de Células , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento/genética , Regulação da Expressão Gênica no Desenvolvimento/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Biblioteca Gênica , Genes de Plantas/genética , Laminaria/genética , Células Fotorreceptoras/metabolismo , Células Fotorreceptoras/efeitos da radiação , Fotossíntese , Proteínas de Plantas/genética , Proteínas Ribossômicas/genética , Regulação para Cima/efeitos da radiação
15.
Int J Mol Sci ; 20(9)2019 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-31083348

RESUMO

Detrimental health consequences from exposure to space radiation are a major concern for long-duration human exploration missions to the Moon or Mars. Cellular responses to radiation are expected to be heterogeneous for space radiation exposure, where only high-energy protons and other particles traverse a fraction of the cells. Therefore, assessing DNA damage and DNA damage response in individual cells is crucial in understanding the mechanisms by which cells respond to different particle types and energies in space. In this project, we identified a cell-specific signature for radiation response by using single-cell transcriptomics of human lymphocyte subpopulations. We investigated gene expression in individual human T lymphocytes 3 h after ex vivo exposure to 2-Gy gamma rays while using the single-cell sequencing technique (10X Genomics). In the process, RNA was isolated from ~700 irradiated and ~700 non-irradiated control cells, and then sequenced with ~50 k reads/cell. RNA in each of the cells was distinctively barcoded prior to extraction to allow for quantification for individual cells. Principal component and clustering analysis of the unique molecular identifier (UMI) counts classified the cells into three groups or sub-types, which correspond to CD4+, naïve, and CD8+/NK cells. Gene expression changes after radiation exposure were evaluated using negative binomial regression. On average, BBC3, PCNA, and other TP53 related genes that are known to respond to radiation in human T cells showed increased activation. While most of the TP53 responsive genes were upregulated in all groups of cells, the expressions of IRF1, STAT1, and BATF were only upregulated in the CD4+ and naïve groups, but were unchanged in the CD8+/NK group, which suggests that the interferon-gamma pathway does not respond to radiation in CD8+/NK cells. Thus, single-cell RNA sequencing technique was useful for simultaneously identifying the expression of a set of genes in individual cells and T lymphocyte subpopulation after gamma radiation exposure. The degree of dependence of UMI counts between pairs of upregulated genes was also evaluated to construct a similarity matrix for cluster analysis. The cluster analysis identified a group of TP53-responsive genes and a group of genes that are involved in the interferon gamma pathway, which demonstrate the potential of this method for identifying previously unknown groups of genes with similar expression patterns.


Assuntos
Exposição à Radiação , Fator de Transcrição STAT1/metabolismo , Análise de Sequência de RNA , Transdução de Sinais , Análise de Célula Única , Subpopulações de Linfócitos T/metabolismo , Subpopulações de Linfócitos T/efeitos da radiação , Proteína Supressora de Tumor p53/metabolismo , Análise por Conglomerados , Raios gama , Humanos , Imunofenotipagem , Reprodutibilidade dos Testes , Transdução de Sinais/genética , Transdução de Sinais/efeitos da radiação , Regulação para Cima/genética , Regulação para Cima/efeitos da radiação
16.
Arch Dermatol Res ; 311(7): 535-544, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31089877

RESUMO

WNT/ß-catenin signaling pathways play a pivotal role in the human immune defense against infections and in chronic inflammatory conditions as psoriasis. Wnt gene alterations are linked to known comorbidities of psoriasis as obesity, diabetes and Crohn's disease. The objective of this study was to investigate WNT7B, WNT10B, WNT16 and TCF7L2 gene and protein expression in lesional and non-lesional skin and in the peripheral blood of patients with chronic plaque psoriasis compared with healthy individuals. To investigate the effect of narrowband UVB radiation, expression of these genes were analyzed before and after narrowband UVB treatment. Associations between single nucleotide polymorphisms for WNT7B, WNT10B, WNT16 and TCF7L2 genes and psoriasis were tested. Our results show significantly decreased WNT7B, WNT10B and TCF7L2 gene expression in lesional skin compared with non-lesional skin and healthy controls. Narrowband UVB treatment significantly increased expression of these genes in lesional skin. Immunohistochemistry shows increased WNT16 expression in lesional skin. No significant differences in allele or genotype frequencies for Wnt or TCF7L2 gene polymorphisms were found between patient and control group. This study shows for the first time significant UVB induced upregulation of WNT7B, WNT10B and TCF7L2 in patients with psoriasis and suggests a potential role of these genes in psoriasis pathogenesis.


Assuntos
Proteínas Proto-Oncogênicas/metabolismo , Psoríase/patologia , Proteína 2 Semelhante ao Fator 7 de Transcrição/metabolismo , Terapia Ultravioleta/métodos , Proteínas Wnt/metabolismo , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Frequência do Gene , Técnicas de Genotipagem , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Proteínas Proto-Oncogênicas/análise , Proteínas Proto-Oncogênicas/genética , Psoríase/genética , Psoríase/radioterapia , Pele/patologia , Pele/efeitos da radiação , Proteína 2 Semelhante ao Fator 7 de Transcrição/análise , Proteína 2 Semelhante ao Fator 7 de Transcrição/genética , Regulação para Cima/efeitos da radiação , Proteínas Wnt/análise , Proteínas Wnt/genética
17.
Mol Oncol ; 13(5): 1249-1267, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30938061

RESUMO

Radioresistance of tumor cells gives rise to local recurrence and disease progression in many patients. MicroRNAs (miRNAs) are master regulators of gene expression that control oncogenic pathways to modulate the radiotherapy response of cells. In the present study, differential expression profiling assays identified 16 deregulated miRNAs in acquired radioresistant breast cancer cells, of which miR-122 was observed to be up-regulated. Functional analysis revealed that miR-122 has a role as a tumor suppressor in parental cells by decreasing survival and promoting radiosensitivity. However, in radioresistant cells, miR-122 functions as an oncomiR by promoting survival. The transcriptomic landscape resulting from knockdown of miR-122 in radioresistant cells showed modulation of the ZNF611, ZNF304, RIPK1, HRAS, DUSP8 and TNFRSF21 genes. Moreover, miR-122 and the set of affected genes were prognostic factors in breast cancer patients treated with radiotherapy. Our data indicate that up-regulation of miR-122 promotes cell survival in acquired radioresistant breast cancer and also suggest that miR-122 differentially controls the response to radiotherapy by a dual function as a tumor suppressor an and oncomiR dependent on cell phenotype.


Assuntos
Neoplasias da Mama , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Genes Supressores de Tumor , MicroRNAs/biossíntese , RNA Neoplásico/biossíntese , Tolerância a Radiação , Regulação para Cima/efeitos da radiação , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Neoplasias da Mama/radioterapia , Feminino , Humanos , Células MCF-7 , MicroRNAs/genética , Proteínas de Neoplasias , RNA Neoplásico/genética
18.
J Dermatol Sci ; 94(2): 276-283, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30956030

RESUMO

BACKGROUND: Ultraviolet B (UVB) radiation is a major cause of skin photodamage, including the damage associated with photodermatoses, aging, and cancer. Although many studies have shown that red light has photoprotective effects on skin, the mechanisms underlying these effects are still poorly understood. OBJECTIVE: The aim of this study was to identify the photoprotective effects of visible red light against UVB-induced skin damage in normal human dermal fibroblast cells using a transcriptomic approach. METHODS: Next-generation sequencing-based transcriptomic analyses were used to profile transcriptomic alterations and identify genes that are differentially expressed by visible red light and by UVB exposure. To understand the biological networks among identified genes, a literature-based biological pathway analysis was performed. Quantitative real-time polymerase chain reaction assays were used for mRNA-level validation of selected key genes. RESULTS: We observed that visible red light contributes to skin cell protection against UVB by modulating gene expression that enhances the adaptive response to redox and inflammatory balancing and by upregulating genes involved in DNA excision repair processes. We also identified that several key genes in the red light-induced biological network were differentially regulated. CONCLUSIONS: Visible red light enhanced the UVB-protective effects in normal human skin cells via the transcriptomic modulation of genes involved in cell-protective processes. Our findings from this next-generation sequencing analysis may lead to a better understanding of the cytoprotective effects of visible red light and provide direction for further molecular or mechanistic studies.


Assuntos
Reparo do DNA/genética , Fibroblastos/efeitos da radiação , Luz , Pele/efeitos da radiação , Raios Ultravioleta/efeitos adversos , Linhagem Celular , Dano ao DNA/efeitos da radiação , Reparo do DNA/efeitos da radiação , Fibroblastos/metabolismo , Perfilação da Expressão Gênica , Humanos , RNA Mensageiro/metabolismo , Transdução de Sinais/genética , Transdução de Sinais/efeitos da radiação , Pele/citologia , Regulação para Cima/efeitos da radiação
19.
Int J Oncol ; 54(4): 1466-1480, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30968148

RESUMO

It is well-known that the activation status of the P53, signal transducer and activator of transcription (Stat)3 and nuclear factor (NF)­κB signaling pathways determines the radiosensitivity of cancer cells. However, the function of these pathways in radiosensitive vs radioresistant cancer cells remains elusive. The present study demonstrated that adaptive expression of epidermal growth factor (EGF) following exposure to ionizing radiation (IR) may induce radiosensitization of pancreatic cancer (PC) cells through induction of the cyclin D1/P53/poly(ADP­ribose) polymerase pathway. By contrast, adaptively expressed interleukin (IL)­6 and insulin­like growth factor (IGF)­1 may promote radioresistance of PC cells, likely through activation of the Stat3 and NF­κB pathways. In addition, cyclin D1 and survivin, which are specifically expressed in the G1/S and G2/M phase of the cell cycle, respectively, are mutually exclusive in radiosensitive and radioresistant PC cells, while Bcl­2 and Bcl­xL expression does not differ between radiosensitive and radioresistant PC cells. Therefore, adaptively expressed EGF and IL­6/IGF­1 may alter these pathways to promote the radiosensitivity of PC cancers. The findings of the present study highlight potential makers for the evaluation of radiosensitivity and enable the development of effective regimens for cancer radiotherapy.


Assuntos
Carcinoma Ductal Pancreático/radioterapia , Fator de Crescimento Epidérmico/metabolismo , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Neoplasias Pancreáticas/radioterapia , Transdução de Sinais/efeitos da radiação , Apoptose/efeitos da radiação , Carcinoma Ductal Pancreático/patologia , Linhagem Celular Tumoral , Ciclina D1/metabolismo , Humanos , Neoplasias Pancreáticas/patologia , Poli(ADP-Ribose) Polimerase-1/metabolismo , Tolerância a Radiação , Proteína Supressora de Tumor p53/metabolismo , Regulação para Cima/efeitos da radiação
20.
DNA Repair (Amst) ; 78: 37-44, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30954901

RESUMO

DNA double strand breaks (DSBs) are a severe threat to genome integrity and a potential cause of tumorigenesis, which is a multi-stage process and involves many factors including the mutation of oncogenes and tumor suppressors, some of which are transcribed microRNAs (miRNAs). Among more than 2000 known miRNAs, miR-21 is a unique onco-miRNA that is highly expressed in almost all types of human tumors and is associated with tumorigenesis through its multiple targets. However, it remains unclear whether there is any functional link between DSBs and miR-21 expression and, if so, does the link contribute to DSB-induced genomic instability/tumorigenesis. To address this question, we used DNA-PKcs-/- (deficient in non-homologous end-joining (NHEJ)) and Rad54-/- (deficient in homologous recombination repair (HRR)) mouse embryonic fibroblasts (MEFs) since NHEJ and HRR are the major pathways for DSB repair in mammalian cells. Our results indicate that levels of miR-21 are elevated in these DSB repair (DSBR) deficient cells, and ionizing radiation (IR) further increases these levels in both wild-type (WT) and DSBR-deficient cells. Interestingly, IR stimulated growth in soft agar and this effect was greatly reduced by blocking miR-21 expression in both WT and DSBR-deficient cells. Taken together, our results suggest that either IR or DSBR-deficient can lead to an upregulation of miR-21 levels and that miR-21 is associated with IR-induced cell growth in soft agar. These results may help our understanding of DSB-induced tumorigenesis and provide information that could facilitate the development of new strategies to prevent DSB-induced carcinogenesis.


Assuntos
Ágar , Quebras de DNA de Cadeia Dupla/efeitos da radiação , Reparo do DNA/efeitos da radiação , MicroRNAs/genética , Regulação para Cima/efeitos da radiação , Animais , Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Carcinogênese/genética , Carcinogênese/efeitos da radiação , Linhagem Celular , Proliferação de Células/genética , Proliferação de Células/efeitos da radiação , Reparo do DNA/genética , Receptores ErbB/metabolismo , Camundongos
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