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1.
Radiat Res ; 195(1): 38-46, 2021 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-33181834

RESUMO

In the event of a mass casualty radiological or nuclear scenario, it is important to distinguish between the unexposed (worried well), low-dose exposed individuals and those developing the hematological acute radiation syndrome (HARS) within the first three days postirradiation. In previous baboon studies, we identified altered gene expression changes after irradiation, which were predictive for the later developing HARS severity. Similar changes in the expression of four of these genes were observed using an in vitro human whole blood model. However, these studies have provided only limited information on the time frame of the changes after exposure in relationship to the development of HARS. In this study we analyzed the time-dependent changes in mRNA expression after in vitro irradiation of whole blood. Changes in the expression of informative mRNAs (FDXR, DBB2, POU2AF1 and WNT3) were determined in the blood of eight healthy donors (6 males, 2 females) after irradiation at 0 (control), 0.5, 2 and 4 Gy using qRT-PCR. FDXR expression was significantly upregulated (P < 0.001) 4 h after ≥0.5 Gy irradiation, with an 18-40-fold peak attained 4-12 h postirradiation which remained elevated (4-9-fold) at 72 h. DDB2 expression was upregulated after 4 h (fold change, 5-8, P < 0.001 at ≥ 0.5 Gy) and remained upregulated (3-4-fold) until 72 h (P < 0.001). The earliest time points showing a significant downregulation of POU2AF1 and WNT3 were 4 h (fold change = 0.4, P = 0.001, at 4 Gy) and 8 h (fold change = 0.3-0.5, P < 0.001, 2-4 Gy), respectively. These results indicate that the diagnostic window for detecting HARS-predictive changes in gene expression may be opened as early as 2 h for most (75%) and at 4 h postirradiation for all individuals examined. Depending on the RNA species studied this may continue for at least three days postirradiation.


Assuntos
Síndrome Aguda da Radiação/diagnóstico , Regulação da Expressão Gênica/efeitos da radiação , RNA Mensageiro/genética , Irradiação Corporal Total/efeitos adversos , Síndrome Aguda da Radiação/genética , Síndrome Aguda da Radiação/patologia , Animais , Relação Dose-Resposta à Radiação , Feminino , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica/genética , Humanos , Masculino , Papio/genética , RNA Mensageiro/efeitos da radiação , Doses de Radiação
2.
Can Respir J ; 2020: 1524716, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32831979

RESUMO

Chronic obstructive pulmonary disease (COPD) is due to structural changes and narrowing of small airways and parenchymal destruction (loss of the alveolar attachment as a result of pulmonary emphysema), which all lead to airflow limitation. Extracorporeal shock waves (ESW) increase cell proliferation and differentiation of connective tissue fibroblasts. To date no studies are available on ESW treatment of human bronchial fibroblasts and epithelial cells from COPD and control subjects. We obtained primary bronchial fibroblasts from bronchial biopsies of 3 patients with mild/moderate COPD and 3 control smokers with normal lung function. 16HBE cells were also studied. Cells were treated with a piezoelectric shock wave generator at low energy (0.3 mJ/mm2, 500 pulses). After treatment, viability was evaluated and cells were recultured and followed up for 4, 24, 48, and 72 h. Cell growth (WST-1 test) was assessed, and proliferation markers were analyzed by qRT-PCR in cell lysates and by ELISA tests in cell supernatants and cell lysates. After ESW treatment, we observed a significant increase of cell proliferation in all cell types. C-Kit (CD117) mRNA was significantly increased in 16HBE cells at 4 h. Protein levels were significantly increased for c-Kit (CD117) at 4 h in 16HBE (p < 0.0001) and at 24 h in COPD-fibroblasts (p = 0.037); for PCNA at 4 h in 16HBE (p = 0.046); for Thy1 (CD90) at 24 and 72 h in CS-fibroblasts (p = 0.031 and p = 0.041); for TGFß1 at 72 h in CS-fibroblasts (p = 0.038); for procollagen-1 at 4 h in COPD-fibroblasts (p = 0.020); and for NF-κB-p65 at 4 and 24 h in 16HBE (p = 0.015 and p = 0.0002). In the peripheral lung tissue of a representative COPD patient, alveolar type II epithelial cells (TTF-1+) coexpressing c-Kit (CD117) and PCNA were occasionally observed. These data show an increase of cell proliferation induced by a low dosage of extracorporeal shock waves in 16HBE cells and primary bronchial fibroblasts of COPD and control smoking subjects.


Assuntos
Brônquios/citologia , Diferenciação Celular/efeitos da radiação , Proliferação de Células/efeitos da radiação , Células Epiteliais/efeitos da radiação , Tratamento por Ondas de Choque Extracorpóreas , Fibroblastos/efeitos da radiação , Doença Pulmonar Obstrutiva Crônica/metabolismo , Idoso , Estudos de Casos e Controles , Linhagem Celular , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Colágeno Tipo I/efeitos da radiação , Humanos , Masculino , Pessoa de Meia-Idade , Cultura Primária de Células , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Antígeno Nuclear de Célula em Proliferação/efeitos da radiação , Proteínas Proto-Oncogênicas c-kit/genética , Proteínas Proto-Oncogênicas c-kit/metabolismo , Proteínas Proto-Oncogênicas c-kit/efeitos da radiação , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , RNA Mensageiro/metabolismo , RNA Mensageiro/efeitos da radiação , Fumantes , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo , Fator de Transcrição RelA/efeitos da radiação , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/metabolismo , Fator de Crescimento Transformador beta1/efeitos da radiação
3.
Arch Biochem Biophys ; 690: 108471, 2020 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-32622788

RESUMO

Stilbenes, an active substances closely related to resistance and quality of grapes, are rarely found in natural resources. However its cumulative amount is affected by ultraviolet radiation (UV). The purpose of this study is to screen key genes in biosynthesis of stilbenes Trans-scripusin A and explore its synthetic pathway. We tested content of stilbenes with UHPLC-QQQ-MS2, results revealed that stilbenes accumulation is positively correlated with UV-B exposure time. Then, we performed transcriptome high-throughput sequencing of grapes under treatments. Results shown that 13,906 differentially expressed genes were obtained, which were mainly enriched in three major regions (ribosome, plant-pathogen interaction and biosynthesis of flavonoid). Three genes of trans-scripusin A synthesis pathway key got by combining KEGG annotation and reference gene HsCYP1B1. Phylogenetic analysis showed that SAH genes had high homology with other hydroxylase genes, and distributed in two subgroups. Gene structure analysis showed that SAH genes contained four exons, indicating that gene has low genetic diversity. Chromosome localization revealed that SAH genes were distributed on different chromosomes, in addition, the number of gene pairs between Vitis vinifera and other species was not related to genome size of other species. The expression profiles of SAH genes in different parts of Vitis vinifera L. were analyzed using qRT-PCR analysis, results indicated that expression of SAH genes be specific to fruit part. These paper provide theoretical basis for further study of polyphenols biosynthesis pathway in grape fruits. The study provides novel insights for further understanding quality of grapes response to UV radiation.


Assuntos
Frutas/genética , Regulação da Expressão Gênica de Plantas/efeitos da radiação , RNA Mensageiro/efeitos da radiação , Vitis/genética , Vias Biossintéticas , Cromatografia Líquida de Alta Pressão , Flavonoides/metabolismo , Frutas/metabolismo , Frutas/efeitos da radiação , Ensaios de Triagem em Larga Escala , Conformação de Ácido Nucleico , Filogenia , Polifenóis/metabolismo , RNA-Seq , Ribossomos/metabolismo , Estilbenos/metabolismo , Estresse Fisiológico/genética , Estresse Fisiológico/efeitos da radiação , Espectrometria de Massas em Tandem , Transcriptoma/efeitos da radiação , Raios Ultravioleta , Vitis/metabolismo , Vitis/efeitos da radiação
4.
Folia Med (Plovdiv) ; 62(2): 314-323, 2020 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-32666749

RESUMO

BACKGROUND: Antimicrobial photothermal/photodynamic therapy (PTT/PDT) with indocyanine green (ICG) is an adjuvant therapeutic approach in the treatment of periodontitis. To explore whether PTT/PDT with ICG causes cell death by apoptosis in human gingival fibroblast (HGF) cells, BAX and BCL-2 genes expression as key events for apoptosis were evaluated in this study. MATERIALS AND METHODS: HGF cells were treated with: 1) different concentrations (500-2000 µg/mL) of ICG alone, 2) Diode laser irradiation alone with a fluency of 39.06 J/cm2; 3) PTT/PDT combined different concentrations (500-2000 µg/mL) of ICG with an 808 nm diode laser with a fluency of 39.06 J/cm2, and 4) controls (untreated cells). After that, BAX and BCL-2 messenger RNA levels were evaluated by real-time quantitative reverse transcription PCR. RESULTS: PTT/PDT with 500 µg/mL of ICG caused significant increases in the expression of the BAX gene, with an 8.5-fold increase, which was approximately 7- and 8.5-fold higher than PTT/PDT with ICG for 1500 and 2000 µg/mL of ICG, respectively, indicating induction of apoptosis in HGF cells. ICG (in different test concentrations), diode laser, and PTT/PDT with ICG (1500 and 2000 µg/mL of ICG) treatment displayed insignificant increases in expression levels of BAX (all p>0.05). Our experiments showed an insignificant increase (1.1-1.6-fold) in the expression of BCL-2 after ICG, diode laser, and PTT/PDT with ICG treatment (all p>0.05). CONCLUSIONS: This study suggests that various concentration of ICG can be the diverse expression of BAX responses to PTT/PDT on HGF cells.


Assuntos
Apoptose/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Verde de Indocianina/farmacologia , Periodontite/terapia , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Proteínas Proto-Oncogênicas c-bcl-2/efeitos dos fármacos , Proteína X Associada a bcl-2/efeitos dos fármacos , Anti-Infecciosos/farmacologia , Apoptose/genética , Apoptose/efeitos da radiação , Células Cultivadas , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Humanos , Terapia Fototérmica , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/efeitos da radiação , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/efeitos da radiação , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/efeitos da radiação
5.
Health Phys ; 119(3): 297-305, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32384371

RESUMO

There is increasing evidence that the expression of non-coding RNA and mRNA (messenger RNA) is significantly altered following high-dose ionizing radiation (IR), and their expression may play a critical role in cellular responses to IR. However, the role of non-coding RNA and mRNA in radiation protection, especially in the nervous system, remains unknown. In this study, microarray profiles were used to determine microRNA (miRNA), long non-coding RNA (lncRNA), and mRNA expression in the hypothalamus of mice that were pretreated with amifostine and subsequently exposed to high-dose IR. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed. We found that fewer miRNAs, lncRNAs, and mRNAs were induced by amifostine pre-treatment in exposed mice, which exhibited antagonistic effects compared to IR, indicating that amifostine attenuated the IR-induced effects on RNA profiles. GO and KEGG pathway analyses showed changes in a variety of signaling pathways involved in inflammatory responses during radioprotection following amifostine pre-treatment in exposed mice. Taken together, our study revealed that amifostine treatment altered or attenuated miRNA, lncRNA, and mRNA expression in the hypothalamus of exposed mice. These data provide a resource to further elucidate the mechanisms underlying amifostine-mediated radioprotection in the hypothalamus.


Assuntos
Amifostina/farmacologia , Radioisótopos de Cobalto/efeitos adversos , Raios gama/efeitos adversos , Hipotálamo/efeitos da radiação , MicroRNAs/efeitos da radiação , RNA Longo não Codificante/efeitos da radiação , RNA Mensageiro/efeitos da radiação , Protetores contra Radiação/farmacologia , Transcriptoma/efeitos da radiação , Animais , Hipotálamo/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Análise de Sequência com Séries de Oligonucleotídeos , Irradiação Corporal Total/efeitos adversos
6.
Life Sci Space Res (Amst) ; 24: 1-8, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31987473

RESUMO

In space, multiple unique environmental factors, particularly microgravity and space radiation, pose a constant threat to astronaut health. MicroRNAs (miRNAs) and long noncoding RNAs (lncRNAs) are functional RNAs that play critical roles in regulating multiple cellular processes. To gain insight into the role of non-coding RNAs in response to radiation and microgravity, we analyzed RNA expression profiles in human lymphoblastoid TK6 cells incubated for 24 h under static or rotating conditions to stimulate microgravity in space, after 2-Gy γ-ray irradiation. The expression of 14 lncRNAs and 17 mRNAs (differentially-expressed genes, DEGs) was found to be significantly downregulated under simulated microgravity conditions. In contrast, irradiation upregulated 55 lncRNAs and 56 DEGs, whereas only one lncRNA, but no DEGs, was downregulated. Furthermore, two miRNAs, 70 lncRNAs, and 87 DEGs showed significantly altered expression in response to simulated microgravity after irradiation, and these changes were independently induced by irradiation and simulated microgravity. GO enrichment and KEGG pathway analyses indicated that the associated target genes showed similar patterns to the noncoding RNAs and were suggested to be involved in the immune/inflammatory response including LPS/TLR, TNF, and NF-κB signaling pathways. However, synergistic effects on RNA expression and cellular responses were also observed with a combination of simulated microgravity and irradiation based on microarray and RT-PCR analysis. Together, our results indicate that simulated microgravity and irradiation additively alter expression patterns but synergistically modulate the expression levels of RNAs and their target genes in human lymphoblastoid cells.


Assuntos
Linfócitos/efeitos da radiação , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , RNA Mensageiro/metabolismo , Simulação de Ausência de Peso , Linhagem Celular , Regulação para Baixo/efeitos da radiação , Humanos , Linfócitos/metabolismo , MicroRNAs/efeitos da radiação , Análise em Microsséries , Mapas de Interação de Proteínas , RNA Longo não Codificante/efeitos da radiação , RNA Mensageiro/efeitos da radiação , Radiação Ionizante , Reação em Cadeia da Polimerase em Tempo Real
7.
Endocr J ; 67(2): 231-240, 2020 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-31748432

RESUMO

Exposure to ionized radiation in childhood has been recognized as a risk factor for the development of thyroid cancer and possibly for other thyroid disorders. However, the effects of neonatal radiation exposure on thyroid morphology and functions have never been explored despite its potential importance. One-week-old male Wistar rats were subjected to cervical X-irradiation at 6 and 12 Gy. Animals were examined at the ages of 2, 8 and 18 weeks old. For comparison, 8-week-old rats were cervically X-irradiated at the same doses. Thyroid histology was examined by computer-assisted microscopy to measure areas of colloid and epithelium of thyroid follicles as well as epithelial heights. In rats that received cervical X-irradiation at 1 week old, the colloid size of thyroid follicles decreased at the age of 8 weeks old in a radiation-dose dependent manner. This morphological change was persistently found at 18 weeks old. There were no significant differences in serum total T3 or T4 levels among the groups. Serum TSH levels increased significantly in 8-week-old rats neonatally X-irradiated. Thyroglobulin (Tg) mRNA and protein expressions were significantly decreased in the neonatally-irradiated group while thyroid peroxidase mRNA express increased at 18 weeks old. None of these changes were observed in the rats X-irradiated at 8 weeks old. In conclusion, our results clearly demonstrated that neonatal rat thyroid was sensitive to ionized radiation, developing specific morphological changes characterized by smaller thyroid follicles along with changes in serum TSH levels and Tg expressions in the thyroid tissue.


Assuntos
Iodeto Peroxidase/efeitos da radiação , Tireoglobulina/efeitos da radiação , Glândula Tireoide/efeitos da radiação , Tireotropina/efeitos da radiação , Tiroxina/efeitos da radiação , Tri-Iodotironina/efeitos da radiação , Raios X , Fatores Etários , Animais , Animais Recém-Nascidos , Western Blotting , Relação Dose-Resposta à Radiação , Iodeto Peroxidase/genética , Iodeto Peroxidase/metabolismo , Masculino , Pescoço , RNA Mensageiro/metabolismo , RNA Mensageiro/efeitos da radiação , Doses de Radiação , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Tireoglobulina/genética , Tireoglobulina/metabolismo , Glândula Tireoide/metabolismo , Glândula Tireoide/patologia , Tireotropina/metabolismo , Tiroxina/metabolismo , Tri-Iodotironina/metabolismo
8.
J Cell Mol Med ; 22(12): 6357-6367, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30324649

RESUMO

LncRNAs have been reported to play an important role in various diseases. However, their role in the radiation-induced intestinal injury is unknown. The goal of the present study was to analyse the potential mechanistic role of lncRNAs in the radiation-induced intestinal injury. Mice were divided into two groups: Control (non-irradiated) and irradiated. Irradiated mice were administered 14 Gy of abdominal irradiation (ABI) and were assessed 3.5 days after irradiation. Changes to the jejuna of ABI mice were analysed using RNA-Seq for alterations to both lncRNA and mRNA. These results were validated using qRT-PCR. LncRNAs targets were predicted based on analysis of lncRNAs-miRNAs-mRNAs interaction. 29 007 lncRNAs and 17 142 mRNAs were detected in the two groups. At 3.5 days post-irradiation, 91 lncRNAs and 57 lncRNAs were significantly up- and downregulated respectively. Similarly, 752 mRNAs and 400 mRNAs were significantly up- and downregulated respectively. qRT-PCR was used to verify the altered expression of four lncRNAs (ENSMUST00000173070, AK157361, AK083183, AK038898) and four mRNAs (Mboat1, Nek10, Ccl24, Cyp2c55). Gene ontology and KEGG pathway analyses indicated the predicted genes were mainly involved in the VEGF signalling pathway. This study reveals that the expression of lncRNAs was altered in the jejuna of mice post-irradiation. Moreover, it provides a resource for the study of lncRNAs in the radiation-induced intestinal injury.


Assuntos
Jejuno/efeitos da radiação , MicroRNAs/genética , RNA Longo não Codificante/genética , RNA Mensageiro/genética , Animais , Regulação da Expressão Gênica/efeitos da radiação , Redes Reguladoras de Genes/genética , Redes Reguladoras de Genes/efeitos da radiação , Jejuno/metabolismo , Jejuno/patologia , Camundongos , MicroRNAs/efeitos da radiação , RNA Longo não Codificante/efeitos da radiação , RNA Mensageiro/efeitos da radiação , Radiação , Fator A de Crescimento do Endotélio Vascular/genética
9.
Audiol Neurootol ; 23(3): 173-180, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30300901

RESUMO

Survival of cochlear sensory epithelial cells may be regulated by inhibitor of differentiation-1 (Id1) and the N-methyl-D-aspartic acid (NMDA) receptor. However, it is unclear whether Id1 and the NMDA receptor are involved in the radiation-mediated survival of rat cochlear sensory epithelial cells. Here, we show that the percentage of apoptotic cells increased, the percentage of cells in the S phase decreased, Id1 mRNA and protein expression decreased and the NMDA receptor subtype 2B (NR2B) mRNA and protein level increased in OC1 cells after radiation. Cells infected with the Id1 gene exhibited higher Id1 mRNA and protein levels and lower NR2B mRNA and protein levels than the control cells. In contrast, after transfection of the Id1 siRNA into OC1 cells, Id1 mRNA and protein expression decreased and NR2B mRNA and protein expression increased relative to that of the control group. Additionally, treatment with ifenprodil for 24 h before radiation reduced apoptosis and increased the percentage of cells in the S phase. Our results suggest that Id1 and NR2B might regulate the survival of OC1 cells following radiation.


Assuntos
Células Epiteliais/efeitos da radiação , Proteína 1 Inibidora de Diferenciação/efeitos da radiação , Órgão Espiral/efeitos da radiação , RNA Mensageiro/efeitos da radiação , Receptores de N-Metil-D-Aspartato/efeitos da radiação , Animais , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Cóclea/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Antagonistas de Aminoácidos Excitatórios/farmacologia , Proteína 1 Inibidora de Diferenciação/genética , Proteína 1 Inibidora de Diferenciação/metabolismo , Órgão Espiral/citologia , Órgão Espiral/efeitos dos fármacos , Órgão Espiral/metabolismo , Piperidinas/farmacologia , RNA Mensageiro/metabolismo , RNA Interferente Pequeno , Ratos , Receptores de N-Metil-D-Aspartato/genética , Receptores de N-Metil-D-Aspartato/metabolismo , Fase S/efeitos dos fármacos , Fase S/efeitos da radiação , Transfecção
10.
Int J Mol Sci ; 19(8)2018 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-30096780

RESUMO

Cell therapy is an innovative strategy for tissue repair, since adult stem cells could have limited regenerative ability as in the case of myocardial damage. This leads to a local contractile dysfunction due to scar formation. For these reasons, refining strategy approaches for "in vitro" stem cell commitment, preparatory to the "in vivo" stem cell differentiation, is imperative. In this work, we isolated and characterized at molecular and cellular level, human Amniotic Mesenchymal Stromal Cells (hAMSCs) and exposed them to a physical Extremely Low Frequency Electromagnetic Field (ELF-EMF) stimulus and to a chemical Nitric Oxide treatment. Physically exposed cells showed a decrease of cell proliferation and no change in metabolic activity, cell vitality and apoptotic rate. An increase in the mRNA expression of cardiac and angiogenic differentiation markers, confirmed at the translational level, was also highlighted in exposed cells. Our data, for the first time, provide evidence that physical ELF-EMF stimulus (7 Hz, 2.5 µT), similarly to the chemical treatment, is able to trigger hAMSC cardiac commitment. More importantly, we also observed that only the physical stimulus is able to induce both types of commitments contemporarily (cardiac and angiogenic), suggesting its potential use to obtain a better regenerative response in cell-therapy protocols.


Assuntos
Diferenciação Celular/efeitos da radiação , Proliferação de Células/efeitos da radiação , Células-Tronco Mesenquimais/efeitos da radiação , Medicina Regenerativa , Âmnio/citologia , Âmnio/crescimento & desenvolvimento , Âmnio/efeitos da radiação , Terapia Baseada em Transplante de Células e Tecidos/métodos , Campos Eletromagnéticos , Regulação da Expressão Gênica no Desenvolvimento/efeitos da radiação , Coração/efeitos da radiação , Humanos , Células-Tronco Mesenquimais/citologia , RNA Mensageiro/efeitos da radiação , Radiação não Ionizante
11.
Clin Cancer Res ; 23(11): 2713-2722, 2017 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-28476872

RESUMO

Purpose: The primary cause of death due to head and neck squamous cell carcinoma (HNSCC) is local treatment failure. The goal of this study was to examine this phenomenon using an unbiased approach.Experimental Design: We utilized human papilloma virus (HPV)-negative cell lines rendered radiation-resistant (RR) via repeated exposure to radiation, a panel of HPV-negative HNSCC cell lines and three cohorts of HPV-negative HNSCC tumors (n = 68, 97, and 114) from patients treated with radiotherapy and subjected to genomic, transcriptomic, and proteomic analysis.Results: RR cell lines exhibited upregulation of several proteins compared with controls, including increased activation of Axl and PI3 kinase signaling as well as increased expression of PD-L1. Additionally, inhibition of either Axl or PI3 kinase led to decreased PD-L1 expression. When clinical samples were subjected to RPPA and mRNA expression analysis, PD-L1 was correlated with both Axl and PI3K signaling as well as dramatically associated with local failure following radiotherapy. This finding was confirmed examining a third cohort using immunohistochemistry. Indeed, tumors with high expression of PD-L1 had failure rates following radiotherapy of 60%, 70%, and 50% compared with 20%, 25%, and 20% in the PD-L1-low expression group (P = 0.01, 1.9 × 10-3, and 9 × 10-4, respectively). This finding remained significant on multivariate analysis in all groups. Additionally, patients with PD-L1 low/CD8+ tumor-infiltrating lymphocytes high had no local failure or death due to disease (P = 5 × 10-4 and P = 4 × 10-4, respectively).Conclusions: Taken together, our data point to a targetable Axl-PI3 kinase-PD-L1 axis that is highly associated with radiation resistance. Clin Cancer Res; 23(11); 2713-22. ©2017 AACR.


Assuntos
Antígeno B7-H1/genética , Carcinoma de Células Escamosas/radioterapia , Neoplasias de Cabeça e Pescoço/radioterapia , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas/genética , Receptores Proteína Tirosina Quinases/genética , Idoso , Biomarcadores Tumorais/genética , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Neoplasias de Cabeça e Pescoço/genética , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Linfócitos do Interstício Tumoral , Masculino , Pessoa de Meia-Idade , Papillomaviridae/patogenicidade , Proteômica , RNA Mensageiro/efeitos da radiação , Tolerância a Radiação/genética , Transdução de Sinais/efeitos da radiação , Receptor Tirosina Quinase Axl
12.
Transfusion ; 56(9): 2286-95, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27443848

RESUMO

BACKGROUND: Pathogen inactivation (PI) techniques use ultraviolet (UV) illumination with or without a photosensitizer to destroy pathogen RNA and DNA. Although lacking a nucleus and innate DNA transcription, platelets (PLTs) contain RNA and can synthesize proteins. The impact of PI on PLT protein synthesis and function is unknown; altered synthesis may affect overall PLT quality. In this study we determine to what extent PLT RNA is affected by PI. STUDY DESIGN AND METHODS: In a pool-and-split design, paired apheresis PLT concentrates were treated with riboflavin and UV illumination or were left untreated. PLT total RNA and mRNA amounts specific for glycoproteins (GP)IIIa, GPIIb, and GPIb; α-granule proteins PLT factor (PF)4; osteonectin and thrombospondin (TSP); and housekeeping protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were determined using absorbance and quantitative polymerase chain reaction. RESULTS: After treatment, amounts of all analyzed mRNAs were significantly reduced (p < 0.05), but to different degrees. For GAPDH and PF4, transcripts appeared less susceptible to the treatment, with 70% remaining 1 hour after UV illumination. For GPIIIa and TSP, less than 15% remained after treatment. There was a correlation (R(2) = 0.85) between transcript length and amount of mRNA remaining 1 hour after treatment. Total RNA demonstrated a life span equal to the PLT life span of 10 to 11 days. CONCLUSION: This is the first report of the impact of riboflavin and UV illumination on PLT mRNA. Results suggest that all mRNA present in PLTs is affected by the treatment although the degree of the effect varies among transcripts.


Assuntos
Plaquetas/metabolismo , RNA Mensageiro/genética , Riboflavina/farmacologia , Raios Ultravioleta , Plaquetas/efeitos dos fármacos , Plaquetas/efeitos da radiação , Preservação de Sangue/métodos , Gliceraldeído-3-Fosfato Desidrogenase (Fosforiladora)/análise , Gliceraldeído-3-Fosfato Desidrogenase (Fosforiladora)/genética , Humanos , Integrina beta3/análise , Integrina beta3/genética , Osteonectina/análise , Osteonectina/genética , Fator Plaquetário 4/análise , Fator Plaquetário 4/genética , Glicoproteína IIb da Membrana de Plaquetas/análise , Glicoproteína IIb da Membrana de Plaquetas/genética , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/efeitos da radiação , Trombospondinas/análise , Trombospondinas/genética
13.
Int J Dermatol ; 55(8): 856-63, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26475182

RESUMO

BACKGROUND: The mechanisms responsible for the efficacy of ultraviolet A1 (UVA1) in the treatment of atopic dermatitis (AD) are not fully understood. OBJECTIVES: This study was designed to investigate mRNA expression of thymic stromal lymphopoietin (TSLP), thymus- and activation-regulated chemokine (TARC), interleukin-5 (IL-5), and IL-13 in AD before and after UVA1 therapy, to determine correlations among them, and to examine whether UVA1 influences their expression and whether it is associated with UVA1 efficacy. METHODS: Twenty-five patients with AD underwent medium-dose UVA1 phototherapy. Before and after UVA1, biopsies from acute skin lesions were studied using reverse transcription and real-time polymerase chain reaction. RESULTS: Levels of mRNA TSLP correlated with those of TARC, IL-5, and IL-13, and levels of TARC correlated with those of IL-5 and IL-13, both before and after UVA1. Expression of IL-5 correlated with that of IL-13 only before UVA1. SCORAD (SCORing of Atopic Dermatitis) indices correlated with levels of TARC and IL-5 before irradiation. After UVA1, no mRNA level correlated with the SCORAD index. Phototherapy with UVA1 improved SCORAD values (P < 0.001) and increased expression of TARC (P < 0.05) but did not affect mRNA expression of TSLP, IL-5, or IL-13. CONCLUSIONS: Expression levels of the mediators TSLP, TARC, IL-5, and IL-13 in AD are interrelated. Phototherapy with UVA1 improves SCORAD indices and increases expression of TARC but has no direct effects on the expression of other molecules. It is likely that UVA1 also interferes with or acts via intermediators on the link between IL-5 and IL-13.


Assuntos
Citocinas/metabolismo , Dermatite Atópica/radioterapia , RNA Mensageiro/metabolismo , Terapia Ultravioleta/métodos , Doença Aguda , Adulto , Biomarcadores/metabolismo , Quimiocina CCL17/genética , Quimiocina CCL17/metabolismo , Estudos de Coortes , Citocinas/genética , Dermatite Atópica/diagnóstico , Feminino , Humanos , Interleucina-13/genética , Interleucina-13/metabolismo , Interleucina-5/genética , Interleucina-5/metabolismo , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/efeitos da radiação , Doses de Radiação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Índice de Gravidade de Doença , Estatísticas não Paramétricas , Resultado do Tratamento , Adulto Jovem , Linfopoietina do Estroma do Timo
14.
Orthod Craniofac Res ; 18 Suppl 1: 50-61, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25865533

RESUMO

OBJECTIVES: This study tested whether or not gene expression in human marrow stromal fibroblast (MSF) cells depends on light wavelength and energy density. MATERIALS AND METHODS: Primary cultures of isolated human bone marrow stem cells (hBMSC) were exposed to visible red (VR, 633 nm) and infrared (IR, 830 nm) radiation wavelengths from a light emitting diode (LED) over a range of energy densities (0.5, 1.0, 1.5, and 2.0 Joules/cm2) Cultured cells were assayed for cell proliferation, osteogenic potential, adipogenesis, mRNA and protein content. mRNA was analyzed by microarray and compared among different wavelengths and energy densities. Mesenchymal and epithelial cell responses were compared to determine whether responses were cell type specific. Protein array analysis was used to further analyze key pathways identified by microarrays. RESULT: Different wavelengths and energy densities produced unique sets of genes identified by microarray analysis. Pathway analysis pointed to TGF-beta 1 in the visible red and Akt 1 in the infrared wavelengths as key pathways to study. TGF-beta protein arrays suggested switching from canonical to non-canonical TGF-beta pathways with increases to longer IR wavelengths. Microarrays suggest RANKL and MMP 10 followed IR energy density dose-response curves. Epithelial and mesenchymal cells respond differently to stimulation by light suggesting cell type-specific response is possible. CONCLUSIONS: These studies demonstrate differential gene expression with different wavelengths, energy densities and cell types. These differences in gene expression have the potential to be exploited for therapeutic purposes and can help explain contradictory results in the literature when wavelengths, energy densities and cell types differ.


Assuntos
Fibroblastos/efeitos da radiação , Expressão Gênica/efeitos da radiação , Raios Infravermelhos , Luz , Células-Tronco Mesenquimais/efeitos da radiação , Adipogenia/efeitos da radiação , Técnicas de Cultura de Células , Linhagem Celular , Proliferação de Células/efeitos da radiação , Células Cultivadas , Cor , Relação Dose-Resposta à Radiação , Células Epiteliais/efeitos da radiação , Perfilação da Expressão Gênica , Humanos , Queratinócitos/efeitos da radiação , Metaloproteinase 10 da Matriz/efeitos da radiação , Células-Tronco Mesenquimais/fisiologia , Análise em Microsséries , Osteogênese/efeitos da radiação , Proteínas Proto-Oncogênicas c-akt/efeitos da radiação , Ligante RANK/efeitos da radiação , RNA Mensageiro/efeitos da radiação , Doses de Radiação , Transdução de Sinais/efeitos da radiação , Fator de Crescimento Transformador beta/efeitos da radiação
15.
Sci Rep ; 4: 5103, 2014 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-24869783

RESUMO

A radiofrequency electromagnetic field (RF-EMF) of 1800 MHz is widely used in mobile communications. However, the effects of RF-EMFs on cell biology are unclear. Embryonic neural stem cells (eNSCs) play a critical role in brain development. Thus, detecting the effects of RF-EMF on eNSCs is important for exploring the effects of RF-EMF on brain development. Here, we exposed eNSCs to 1800 MHz RF-EMF at specific absorption rate (SAR) values of 1, 2, and 4 W/kg for 1, 2, and 3 days. We found that 1800 MHz RF-EMF exposure did not influence eNSC apoptosis, proliferation, cell cycle or the mRNA expressions of related genes. RF-EMF exposure also did not alter the ratio of eNSC differentiated neurons and astrocytes. However, neurite outgrowth of eNSC differentiated neurons was inhibited after 4 W/kg RF-EMF exposure for 3 days. Additionally, the mRNA and protein expression of the proneural genes Ngn1 and NeuroD, which are crucial for neurite outgrowth, were decreased after RF-EMF exposure. The expression of their inhibitor Hes1 was upregulated by RF-EMF exposure. These results together suggested that 1800 MHz RF-EMF exposure impairs neurite outgrowth of eNSCs. More attention should be given to the potential adverse effects of RF-EMF exposure on brain development.


Assuntos
Campos Eletromagnéticos , Células-Tronco Embrionárias/efeitos da radiação , Células-Tronco Neurais/efeitos da radiação , Neurogênese/efeitos da radiação , Animais , Apoptose/efeitos da radiação , Telefone Celular , Proliferação de Células/efeitos da radiação , Relação Dose-Resposta à Radiação , Humanos , Camundongos , Neuritos/efeitos da radiação , RNA Mensageiro/biossíntese , RNA Mensageiro/efeitos da radiação , Ondas de Rádio
16.
Radiat Prot Dosimetry ; 154(1): 37-44, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22923252

RESUMO

Growth arrest DNA damage-inducible 45a gene (Gadd45a) and immediate early response gene 5 (Ier5) have been emphasised as ideal radiation biomarkers in several reports. However, some aspects of radiation-induced transcriptional alterations of these genes are unknown. In this study, gender-dependency and dose-dependency as two factors that may affect radiation-induced transcription of Gadd45a and Ier5 genes were investigated. Human lymphocyte cells from six healthy voluntary blood donors (three women and three men) were irradiated in vitro with doses of 0.5-4.0 Gy from a (60)Co source and RNA isolated 4 h later using the High Pure RNA Isolation Kit. Dose and gender dependency of radiation-induced transcriptional alterations of Gadd45a and Ier5 genes were studied by quantitative real-time polymerase chain reaction. The results showed that as a whole, Gadd45a and Ier5 gave responses to gamma rays, while the responses were independent of radiation doses. Therefore, regardless of radiation dose, Gadd45a and Ier5 can be considered potential radiation biomarkers. Besides, although radiation-induced transcriptional alterations of Gadd45a in female and male lymphocyte samples were insignificant at 0.5 Gy, at other doses, their quantities in female samples were at a significantly higher level than in male samples. Radiation-induced transcription of Ier5 of females samples had a reduction in comparison with male samples at 1 and 2 Gy, but at doses of 0.5 and 4 Gy, females were significantly more susceptible to radiation-induced transcriptional alteration of Ier5.


Assuntos
Proteínas de Ciclo Celular/genética , Radioisótopos de Cobalto/efeitos adversos , Raios gama/efeitos adversos , Proteínas Imediatamente Precoces/genética , Linfócitos/efeitos da radiação , Proteínas Nucleares/genética , RNA Mensageiro/efeitos da radiação , Transcrição Gênica/efeitos da radiação , Adulto , Biomarcadores/sangue , Relação Dose-Resposta à Radiação , Feminino , Identidade de Gênero , Humanos , Masculino , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real
17.
Endocrinology ; 152(12): 4525-36, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22045660

RESUMO

PTH stimulates bone formation and increases hematopoietic stem cells through mechanisms as yet uncertain. The purpose of this study was to identify mechanisms by which PTH links actions on cells of hematopoietic origin with osteoblast-mediated bone formation. C57B6 mice (10 d) were nonlethally irradiated and then administered PTH for 5-20 d. Irradiation reduced bone marrow cellularity with retention of cells lining trabeculae. PTH anabolic activity was greater in irradiated vs. nonirradiated mice, which could not be accounted for by altered osteoblasts directly or osteoclasts but instead via an altered bone marrow microenvironment. Irradiation increased fibroblast growth factor 2, TGFß, and IL-6 mRNA levels in the bone marrow in vivo. Irradiation decreased B220 cell numbers, whereas the percent of Lin(-)Sca-1(+)c-kit(+) (LSK), CD11b(+), CD68(+), CD41(+), Lin(-)CD29(+)Sca-1(+) cells, and proliferating CD45(-)Nestin(+) cells was increased. Megakaryocyte numbers were reduced with irradiation and located more closely to trabecular surfaces with irradiation and PTH. Bone marrow TGFß was increased in irradiated PTH-treated mice, and inhibition of TGFß blocked the PTH augmentation of bone in irradiated mice. In conclusion, irradiation created a permissive environment for anabolic actions of PTH that was TGFß dependent but osteoclast independent and suggests that a nonosteoclast source of TGFß drives mesenchymal stem cell recruitment to support PTH anabolic actions.


Assuntos
Medula Óssea/efeitos da radiação , Microambiente Celular/efeitos da radiação , Hormônio Paratireóideo/metabolismo , Animais , Contagem de Células , Células-Tronco Mesenquimais/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Osteoclastos , Hormônio Paratireóideo/administração & dosagem , RNA Mensageiro/efeitos da radiação , Fator de Crescimento Transformador beta/fisiologia , Irradiação Corporal Total
18.
Radiother Oncol ; 101(1): 233-6, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21724286

RESUMO

BACKGROUND AND PURPOSE: Systems are being developed to assess radiation exposure based on leukocyte mRNA levels obtained by finger-stick sampling. The goal is to provide accurate detection of dose exposures up to 10 Gy for up to 1 week following exposure. We previously showed that specific mRNA sequences increase expression within an hour of exposure, and some genes continue to show elevated expression for at least 24 h. Full duration and dose-dependence of this persistence remain to be determined. In the present study, real-time quantitative PCR (qPCR) was used to determine changes in gene expression. qPCR can rapidly analyze small blood samples and could be adopted into a field-portable instrument that provides a radiation dose readout within 30 min. MATERIALS AND METHODS: From previous microarray analysis of 21,000 genes expressed in human lymphoblastoid cells 4 h post-irradiation (0-4 Gy), 118 genes were selected for evaluation by qPCR of gene expression in the leukocytes of human blood irradiated in vitro with doses of 0-10 Gy from a Co-60 gamma source at a dose rate of 30 cGy/min. RESULTS: Blood from 20 normal healthy human donors yielded many mRNA sequences that could be used for radiation dosimetry. We observed four genes with large and persistent responses following exposure: ASTN2, CDKN1A, GADD45A, and GDF15. Five genes were identified as reliably non-responsive and were suitable for use as endogenous controls: DPM1, ITFG1, MAP4, PGK1, and SLC25A36; of these, ITFG1 was used for the analyses presented here. A significant dose-responsive increase in expression occurred for CDKN1A that was >16-fold at 10 Gy and 3-fold at 0.5 Gy compared to pre-irradiation values. CONCLUSIONS: These data show large, selective increases in mRNA transcript levels that persist for at least 48 h after single exposures between 0.5 and 10 Gy. Stable, non-responsive mRNA sequences for use as endogenous controls were also identified. These results indicate that following further study to establish the most reproducible gene and dose-response models under a wide range of conditions in vivo, rapid real-time qPCR on blood samples could potentially be used to establish biologically-effective dosimetry from either accidental irradiation or clinical radiotherapy.


Assuntos
Ensaios de Triagem em Larga Escala , Linfócitos/efeitos da radiação , Sistemas Automatizados de Assistência Junto ao Leito , Reação em Cadeia da Polimerase/métodos , RNA Mensageiro/efeitos da radiação , Doses de Radiação , Células Cultivadas/efeitos da radiação , Relação Dose-Resposta à Radiação , Feminino , Raios gama , Perfilação da Expressão Gênica , Humanos , Masculino , Análise de Sequência com Séries de Oligonucleotídeos , Tolerância a Radiação/genética , Sensibilidade e Especificidade
19.
Health Phys ; 101(3): 259-64, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21799342

RESUMO

To examine the effects of carbon ion and gamma ray irradiation on cancer-induced osteoclastogenesis, mouse calvaria MC3T3-E1 cells were cultured with conditioned medium from irradiated and non-irradiated MCF7 human breast cancer cells. The authors examined RANKL and OPG mRNA expression in osteoblastic MC3T3-E1 cells following treatment with conditioned MCF7 medium. Co-cultured MC3T3-E1 and bone marrow cells treated with conditioned medium from irradiated MCF7 cells showed decreased numbers of osteoclasts, assessed using TRAP staining. Conditioned medium from control MCF7 cells elevated the RANKL/OPG mRNA ratio in MC3T3-E1 cells; this effect was suppressed by carbon ion irradiation of the MCF7 cells. These data demonstrate that indirect interactions between breast cancer cells and MC3T3-E1 cells induce osteoclastogenesis in vitro through modulation of RANKL expression and that this process is suppressed by carbon ion irradiation.


Assuntos
Calo Ósseo/efeitos da radiação , Neoplasias da Mama/patologia , Carbono/uso terapêutico , Raios gama/uso terapêutico , Osteogênese/efeitos da radiação , Células 3T3 , Animais , Neoplasias Ósseas/etiologia , Neoplasias Ósseas/metabolismo , Neoplasias Ósseas/patologia , Neoplasias da Mama/metabolismo , Feminino , Regulação da Expressão Gênica/efeitos da radiação , Radioterapia com Íons Pesados , Humanos , Camundongos , RNA Mensageiro/metabolismo , RNA Mensageiro/efeitos da radiação , Receptor Ativador de Fator Nuclear kappa-B/metabolismo , Receptor Ativador de Fator Nuclear kappa-B/efeitos da radiação
20.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 27(1): 15-8, 2011 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-21208556

RESUMO

AIM: To evaluate the effects of electromagnetic irradiation of 2000 µW/cm(2); exposure on mRNA and protein expression levels of immunoreactive protein and mRNA of NMDA receptor 2A subunit in rats hippocampal, and to explore the mechanism of electromagnetic irradiation induced learning and memory impairment. METHODS: Rats were randomly divided into normal control group, sham-radiated group, and 1 h/d, 2 h/d, and 3 h/d radiation groups. The rats in the radiation groups were fixed after microwave exposure of 2000 µW/cm(2);, then their learning and memory abilities were tested by Morris water maze experiment, the change of NR2A protein in hippocampal neurons of each group of rats were measured with immunohistochmistry and Western blot techniques, and the expression of NR2A mRNA in hippocampus were determined by RT-PCR. RESULTS: Compared with the normal control group, each index of the sham-radiated group has no significant change (P>0.05), while the latency of rats of radiated group in Morris water maze test were significantly longer (P<0.05). In the radiation group, the hippocampal neurons of rats showing evident reduction in the ratio of NR2A positive cells, irregular, and arrayed in disorder. Moreover, the expession of NR2A protein and its mRNA in hippocampal neurons were significant decreased (P<0.05). CONCLUSION: Electromagnetic irradiation of 2000 µW/cm(2); exposure can impair the learning and memory abilities of rats possibly through a mechanism correlated with the lower expression of NR2A protein and its mRNA in hippocampus.


Assuntos
Hipocampo/metabolismo , Hipocampo/efeitos da radiação , Micro-Ondas , Radiação , Receptores de N-Metil-D-Aspartato/genética , Receptores de N-Metil-D-Aspartato/efeitos da radiação , Animais , Aprendizagem/efeitos da radiação , Masculino , Memória/efeitos da radiação , Neurônios/metabolismo , Neurônios/efeitos da radiação , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , RNA Mensageiro/efeitos da radiação , Ratos , Ratos Wistar , Receptores de N-Metil-D-Aspartato/biossíntese
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