RESUMO
The aim of this study was to determine the toxicity induced by irradiation with alpha-particles on malignant transformation of immortalized human bronchial epithelial cells (BEAS-2B) using miRNA-mRNA networks. The expression of BEAS-2B cells was determined by measuring colony formation, mtDNA, mitochondrial membrane potential (MMP), and ROS levels. Changes in BEAS-2B cell gene expression were observed and quantified using microarrays that included an increase in 157 mRNA and 20 miRNA expression and a decrease in 77 mRNA and 48 miRNA. Bioinformatic software was used to analyze these different mRNA and miRNA, which indicated that miR-107 and miR-494 play an important role in alpha-particles-mediated cellular malignant transformation processes. The pathways related to systemic lupus erythematosus, cytokine-cytokine receptor interaction, MAPK signaling pathway, regulation of actin cytoskeleton, and cell adhesion molecules (CAMs) were stimulated, while those of ribosome, transforming growth factor (TGF)-beta signaling pathway, and metabolic pathways were inhibited. Data suggest that miRNA and mRNA play a crucial role in alpha-particles-mediated malignant transformation processes. It is worth noting that three target genes associated with lung cancer were identified and upregulated PEG 10 (paternally expressed gene 10), ARHGAP26, and IRS1.
Assuntos
Partículas alfa/efeitos adversos , Transformação Celular Neoplásica/efeitos da radiação , Células Epiteliais/efeitos da radiação , MicroRNAs/metabolismo , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos da radiação , Brônquios/efeitos da radiação , Linhagem Celular , Transformação Celular Neoplásica/metabolismo , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Humanos , Neoplasias Pulmonares/etiologiaRESUMO
Radon and its progeny are confirmed to be type I carcinogenic agents accounting for increased risks in 10% of observed lung cancers globally. However, the underlying carcinogenic mechanisms are largely unknown. In the present study, BEAS2B cells were directly exposed twice to 20,000 Bq/m(3) radon gas for 20 min once (first passage) and subsequently 10 times (fifth passage). The fifth-passage cells were then subcultured for 1 and 20 generations (named Rn5-1 and Rn5-20, respectively). Molecular mechanisms indicative of malignant transformation were assessed by determination of apoptosis, seroresistance, and microRNA (miRNA) expression profiles. The microRNA profiles were used to assess the functional annotations of the target genes. Data indicated an increased seroresistance and colony efficiency on soft agar, and enhanced apoptosis resistance in the Rn5-20 cells with significant differential expressions in some miRNA, including hsa-miR-483-3p, hsa-miR-494, hsa-miR-2115*, hsa-miR-33b, hsa-miR-1246, hsa-miR-3202, hsa-miR-18a, hsa-miR-125b, hsa-miR-17*, and hsa-miR-886-3p. Functional annotation demonstrated that these miRNA target genes were predominantly involved in the regulation of cell proliferation, differentiation, and adhesion during the process of malignant transformation, which is associated with signal pathways such as mitogen-activated protein kinase (MAPK), Int and Wg (Wnt), reactive oxygen species (ROS), nuclear factor κB (NF-κB), and other genes regulating cell cycles.
Assuntos
Poluentes Radioativos do Ar/toxicidade , Carcinógenos Ambientais/toxicidade , Transformação Celular Neoplásica/induzido quimicamente , MicroRNAs/metabolismo , Radônio/toxicidade , Transcriptoma/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Linhagem Celular Transformada , Proliferação de Células/efeitos dos fármacos , Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/patologia , Perfilação da Expressão Gênica , Humanos , Análise de Sequência com Séries de OligonucleotídeosRESUMO
Oxidative damage can be induced by many environmental stressors. 8-Hydroxydeoxyguanosine (8-OHdG) has been used as a biomarker of oxidative DNA damage in both in vitro and in vivo studies. In the present study, Wistar rats were exposed to radon gas at a concentration of 100,000Bq/m(3) for 12 h/d for 30, 60, and 120 d, equivalent to cumulative doses of 60, 120, and 240 working level months (WLM), respectively. Changes in levels of 8-OHdG, reactive oxygen species (ROS), and total antioxidant (T-AOC), as well as expressions of some DNA repair enzymes such as 8-oxoguanine DNA glycosylase (OGG1) and MutT homolog 1 (oxidized purine nucleoside triphosphatase, MTH1), were determined in rat urine, peripheral blood lymphocytes, and lung after exposure to radon. The results revealed an increase in 8-OHdG and ROS levels, a decrease in T-AOC levels, and reduced OGG1 and MTH1 expression levels. The elevated amount of 8-OHdG in urine or lymphocytes was positively correlated with the cumulative exposure dose, whereas OGG1 and MHT1 expression levels in lung were inversely correlated with cumulative exposure dose. These findings indicate that oxidative damage induced by radon may be involved in radon-induced carcinogenesis.
Assuntos
Poluentes Radioativos do Ar/toxicidade , Antioxidantes/metabolismo , Desoxiguanosina/análogos & derivados , Estresse Oxidativo/efeitos da radiação , Radônio/toxicidade , Espécies Reativas de Oxigênio/metabolismo , 8-Hidroxi-2'-Desoxiguanosina , Animais , Biomarcadores , Líquido da Lavagem Broncoalveolar/citologia , DNA Glicosilases/genética , DNA Glicosilases/metabolismo , Reparo do DNA , Enzimas Reparadoras do DNA/metabolismo , Desoxiguanosina/metabolismo , Regulação Enzimológica da Expressão Gênica/fisiologia , Pulmão/metabolismo , Masculino , Neoplasias Induzidas por Radiação/etiologia , Neoplasias Induzidas por Radiação/metabolismo , Monoéster Fosfórico Hidrolases/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos WistarRESUMO
In the present study, we investigated the role of UHRF1 (ubiquitin-like protein containing PHD and RING finger domains 1) in proliferation, invasion and migration of breast cancer cells, and the potential mechanisms were also explored. Cell proliferation was examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay; cell cycle distribution and apoptosis were evaluated using flow cytometry; protein expression was determined by western blotting; angiogenesis of xenografts was assessed by microvessel density (MVD); cell invasion was measured using transwell chamber; cell migration was determined by wound scratching assay. Our results demonstrated that UHRF1 transfection conferred serum independence to MDA-MB-231 cells, G1 phase shortage and apoptosis suppression, accompanied with an increased expression of cyclin D1 and decreased expression of Bax. Significant pro-invasion and pro-migration activity was observed, with no obvious effect on the expression of PTEN and maspin. Co-expression of the UHRF1/PTEN or UHRF1/maspin degraded the role of UHRF1 in regulating invasion and migration. UHRF1 induced growth of MDA-MB-231 cells by promoting tumor vessel formation in vivo. In conclusion, UHRF1 promoted the proliferation of breast cancer cells by apoptosis inhibition, G1 phase shortage and promotion of tumor vessel formation, and pro-invasion and pro-migration activity was also observed by interacting with PTEN and maspin. Thus, UHRF1 may serve as a new therapy target for breast cancer.
Assuntos
Adenocarcinoma/secundário , Neoplasias da Mama/patologia , Proteínas Estimuladoras de Ligação a CCAAT/fisiologia , Adenocarcinoma/irrigação sanguínea , Adenocarcinoma/metabolismo , Animais , Apoptose , Proteínas Reguladoras de Apoptose/metabolismo , Neoplasias da Mama/irrigação sanguínea , Neoplasias da Mama/metabolismo , Proteínas Estimuladoras de Ligação a CCAAT/genética , Proteínas Estimuladoras de Ligação a CCAAT/metabolismo , Pontos de Checagem do Ciclo Celular , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Sobrevivência Celular , Feminino , Expressão Gênica , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Invasividade Neoplásica , Transplante de Neoplasias , Neovascularização Patológica/metabolismo , Carga Tumoral , Ubiquitina-Proteína LigasesRESUMO
PURPOSE: To investigate whether an adaptive response can be induced in mice which were pre-exposed to 900 MHz radiofrequency fields. MATERIALS AND METHODS: Adult male Kunming mice were exposed to 900 MHz radiofrequency fields (RF) at power intensities of 12, 120 and 1200 µW/cm(2) for 1 h/day for 14 days and then subjected to whole body gamma-irradiation. The results were compared with those in unexposed control animals and those exposed to gamma-irradiation alone (without pre-exposure to RF). The extent of survival and hematopoietic tissue damage (assessed in the form of nucleated colony forming cells in the bone marrow and colony forming cells in the spleen of lethally irradiated 'recipient' mice) as well as the expression of cell cycle-related genes were investigated. RESULTS: The results indicated a significant increase in survival time, reduction in the hematopoietic tissue damage in RF pre-exposed mice which were gamma-irradiated (as compared with those exposed to gamma-radiation alone). This was accompanied by significantly increased expression of cell cycle-related genes, namely, cyclin-D1, cyclin-E, cyclin-DK4 and cyclin-DK2 in hematopoietic cells. CONCLUSIONS: Pre-exposure of mice to 900 MHz radiofrequency fields has resulted in a significant reduction in hematopoietic damage caused by subsequent exposure to ionising radiation. This phenomenon appears to be similar to that of the 'adaptive response' which is well documented in scientific literature.
Assuntos
Adaptação Fisiológica/fisiologia , Adaptação Fisiológica/efeitos da radiação , Células-Tronco Hematopoéticas/fisiologia , Células-Tronco Hematopoéticas/efeitos da radiação , Tolerância a Radiação/fisiologia , Tolerância a Radiação/efeitos da radiação , Animais , Sobrevivência Celular/efeitos da radiação , Células Cultivadas , Relação Dose-Resposta à Radiação , Raios gama , Masculino , Camundongos , Doses de Radiação , Irradiação Corporal TotalRESUMO
This study was designed to construct and identify the subtracted cDNA library in peripheral blood cells of BALB/c mice and tracheal-bronchial epithelial cells of Wistar rats following exposure to radon inhalation. Two groups of the animals were exposed in a radon chamber at an accumulative dose of 100 WLM, while control animals were housed in a room at a background dose of 1 WLM. To construct a subtracted cDNA library enriched with differentially expressed genes, the SMART technique and suppression subtractive hybridization (SSH) assay were performed. The obtained forward and reverse cDNA fragments were directly inserted into a pMD-18 vector and transformed into Escherichia coli JM109. In total, 593 white bacterial clones were selected from both forward- and reverse-subtracted libraries. Among them, 81 clones were chosen for their differential expressions based on reverse Northern blot. Portions of these cDNA clones were also verified by a quantitative real-time polymerase chain reaction (PCR). The screening resulted in 14 upregulative and 8 downregulative known function/annotation genes, which were revealed to be functionally related to cell proliferation, cell oxidative and DNA damage, apoptosis, and tumor promotion. Access numbers were obtained from the GenBank for 11 unknown expressed sequence tags (EST). Analysis of biological roles of these cDNA fragments may provide further insight into mechanisms underlying adverse molecular events induced by high-dose radon exposure.
Assuntos
Células Sanguíneas/efeitos da radiação , Células Epiteliais/efeitos da radiação , Expressão Gênica/efeitos da radiação , Radônio/toxicidade , Mucosa Respiratória/efeitos da radiação , Animais , Biomarcadores/metabolismo , Células Sanguíneas/metabolismo , Brônquios/citologia , Brônquios/metabolismo , Brônquios/efeitos da radiação , Relação Dose-Resposta à Radiação , Regulação para Baixo/efeitos da radiação , Células Epiteliais/metabolismo , Biblioteca Gênica , Exposição por Inalação , Masculino , Camundongos , Camundongos Endogâmicos BALB C , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Mucosa Respiratória/citologia , Mucosa Respiratória/metabolismo , Traqueia/citologia , Traqueia/metabolismo , Traqueia/efeitos da radiação , Regulação para Cima/efeitos da radiaçãoRESUMO
Exposure of humans simultaneously to microwave and gamma-ray irradiation may be a commonly encountered phenomenon. In a previous study data showed that low-dose microwave radiation increased the survival rate of mice irradiated with 8Gy gamma-ray; however, the mechanisms underlying these findings remain unclear. Consequently, studies were undertaken to examine the effects of microwave exposure on hematopoietic system adversely altered by gamma-ray irradiation in mice. Preexposure to low-dose microwaves attenuated the damage produced by gamma-ray irradiation as evidenced by less severe pathological alterations in bone marrow and spleen. The protective effects of microwaves were postulated to be due to up-expression of some hematopoietic growth factors, stimulation of proliferation of the granulocyte-macrophages in bone marrow, and inhibition of the gamma-ray induced suppression of hematopoietic stem cells/hematopoietic progenitor cells. Data thus indicate that prior exposure to microwaves may be beneficial in providing protection against injuries produced by gamma-ray on the hematopoietic system in mice.
Assuntos
Raios gama/efeitos adversos , Sistema Hematopoético/efeitos da radiação , Micro-Ondas/uso terapêutico , Lesões Experimentais por Radiação/radioterapia , Proteção Radiológica , Animais , Proliferação de Células/efeitos da radiação , Relação Dose-Resposta à Radiação , Fator Estimulador de Colônias de Granulócitos e Macrófagos/sangue , Células Progenitoras de Granulócitos e Macrófagos/efeitos da radiação , Sistema Hematopoético/metabolismo , Sistema Hematopoético/patologia , Interleucina-3/metabolismo , Masculino , Camundongos , Lesões Experimentais por Radiação/metabolismo , Lesões Experimentais por Radiação/patologia , Baço/patologia , Baço/efeitos da radiaçãoRESUMO
Mobile phones are widely used globally. However, the biological effects due to exposure to electromagnetic fields (EMF) produced by mobile phones are largely unknown. Environmental and occupational exposure of humans to gamma-rays is a biologically relevant phenomenon. Consequently studies were undertaken to examine the interactions between gamma-rays and EMF on human health. In this study, exposure to 900-MHz EMF expanded gamma-ray damage to SHG44 cells. Preexposure EMF enhanced the decrease in cell proliferation induced by gamma-ray irradiation and the rate of apoptosis. The combination of EMF and gamma-ray exposure resulted in a synergistic effect by triggering stress response, which increased reactive oxygen species, but the expression of hsp70 at both mRNA and protein levels remained unaltered. Data indicate that the adverse effects of gamma-rays on cellular functions are strengthened by EMF.
Assuntos
Apoptose/efeitos da radiação , Raios gama/efeitos adversos , Micro-Ondas/efeitos adversos , Neuroglia/efeitos da radiação , Estresse Fisiológico/efeitos da radiação , Ciclo Celular/efeitos da radiação , Linhagem Celular Tumoral , Telefone Celular , Proliferação de Células/efeitos da radiação , HumanosRESUMO
This study examined the protein expression in lung tissues of rats exposed to radon and cigarette smoke using a proteomic approach. Male Wistar rats were exposed daily to radon at a concentration of 100,000 Bq/m(3) for 16 h, and then exposed to 20% cigarette smoke for 1 h for a period of 75 d, with the radon cumulative dose reaching 200 WLM (working level months). Proteins from rat lung tissue were separated by two-dimensional gel electrophoresis (2-DE), stained with Coomassie blue, and analyzed with ImageMaster two-dimensional (2D) platinum software. Differentially expressed proteins were analyzed by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MALDI-time of flight [TOF] MS or MALDI TOF/TOF-MS). Twenty prominent proteins that were correlated with signal transduction, metabolism, heat shock and stress, and cytoskeleton construction were identified. Some of the differential expression proteins were verified by Western blot analysis and immunohistochemical staining, and the results were consistent with 2-DE analysis. The identified proteins and peptides might be potential diagnostic markers of lung impairment induced by radon and cigarette smoke exposure.
Assuntos
Pulmão/efeitos dos fármacos , Pulmão/efeitos da radiação , Radônio/efeitos adversos , Poluição por Fumaça de Tabaco/efeitos adversos , Animais , Biomarcadores/análise , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/efeitos da radiação , Regulação para Baixo , Eletroforese em Gel Bidimensional , Regulação da Expressão Gênica , Pulmão/patologia , Masculino , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/efeitos da radiação , Proteômica , Ratos , Ratos Wistar , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/efeitos da radiação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Regulação para CimaRESUMO
Cigarette smoke has been widely investigated in terms of epidemiology and pathological endpoints in relation to human lung diseases and animal study. In this study we exposed Wistar rats to cigarette smoke at concentrations of 20% and 60% to explore potential molecular mechanisms at the protein level. Exposures were conducted twice a day, 5 days a week for 43 weeks. As a major metabolite of nicotine in cigarette, cotinine level in rat urine was determined by HPLC-MS. A dose-dependent analysis indicated that cotinine may be used as an exposure marker of cigarette smoke. Expression of receptor for advanced glycation endproducts (RAGE), an immunoglobulin super family that triggers the intracellular signal cascade reaction leading to inflammation and its ligand S100A6 (calgranulin) in bronchial epithelial cells and lung tissues of rats, were found to be positive correlated with cotinine levels, indicating that RAGE and S100A6 may be attributable to inflammation and oxidative damage caused by cigarette smoke.
RESUMO
OBJECTIVE: To investigate antagonistic effect of microwave on hematopoietic damage of mice induced by gamma-ray irradiation. METHODS: Male healthy Kunning mice were treated with low dose microwave radiation before exposure to (60)Co gamma-ray irradiation of 8.0 Gy. The 30-day survival rate and average survival time of the mice after the treatment were examined. Peripheral blood parameters and the organ indexes of thymus and spleen were also observed in the irradiated mice. After exposure to 5.0 Gy gamma irradiation, indexes of hematopoietic foci formation of bone marrow cells (CFU-GM) and the proliferation activity of BMNCs were examined. The serum concentration of hemopoietic factors (GM-CSF and IL-3) were detected by ELISA kits. RESULTS: Pre-exposure with 120 microW/cm(2) 900 MHz microwave increased the 30-day survival rate (P < 0.05) and the number of white blood cells of gamma-ray treated mice. The increases of the organ indexes of thymus and spleen, proliferation activity of BMNCs and CFU-GM hematopoietic foci numbers, as well as the higher serum concentration of GM-CSF and IL-3 were observed in the microwave pre-exposure group. CONCLUSION: Low dose microwave radiation may exert potential antagonistic effects on hematopoietic injuries induced by ionizing radiation. The underlying mechanisms might be related with stimulation of hematopoietic growth factors expression, promotion of HSCs/HPCs proliferation, suppression on the reduction of HSCs/HPCs caused by (60)Co gamma-ray, and enhanced construction of the hematopoietic system.
Assuntos
Medula Óssea/efeitos da radiação , Raios gama/efeitos adversos , Micro-Ondas , Lesões Experimentais por Radiação/prevenção & controle , Animais , Medula Óssea/patologia , Células da Medula Óssea/patologia , Células da Medula Óssea/efeitos da radiação , Diferenciação Celular/efeitos da radiação , Proliferação de Células/efeitos da radiação , Fator Estimulador de Colônias de Granulócitos e Macrófagos/sangue , Interleucina-3/sangue , Masculino , Camundongos , Lesões Experimentais por Radiação/sangue , Lesões Experimentais por Radiação/patologiaRESUMO
The aim of this study was to investigate the differential expression of proteins in lung of rats following long-term exposure to radon. The total proteins of lung tissue from Wistar rats exposed to radon for cumulative doses up to 100, 200, or 400 WLM (working level months) were isolated by two-dimensional electrophoresis (2-DE) and analyzed with ImageMaster 2D Platinum software. Comparison of the 2-DE images between the control and radon-exposed groups resulted in 14 upregulated and 9 downregulated protein spots, of which 15 were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) or matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF-MS). The simultaneous up-expressions of RAGE and S100A6 indicated that both proteins might be applied as biomarkers for lung injury induced by long-term radon exposure.
Assuntos
Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Proteoma/metabolismo , Radônio/farmacologia , Animais , Relação Dose-Resposta a Droga , Eletroforese em Gel Bidimensional , Pulmão/patologia , Masculino , Proteômica , Ratos , Ratos Wistar , Receptor para Produtos Finais de Glicação Avançada , Receptores Imunológicos/metabolismo , Proteínas S100/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
This study was designed to screen for differential expression genes in bone marrow cells of mice exposed to radon inhalation. Based upon established pathological findings in mouse, differential screening of gene expressions was conducted by using the SSH method. Among 285 cDNA clones selected from both forward- and reverse subtracted libraries, 45 were chosen for their differential expressions based on reverse Northern blot and quantitative real-time PCR analysis. Of these, up-regulation of the mRNA levels of E-cadherin and down-regulation of the replication protein A1 (RPA1) and casein kinase 1 delta (CKI delta) were also verified by a quantitative real-time PCR. Biological roles of these obtained cDNAs are described and the results of the screening may provide important clues for further investigations of the adverse molecular events induced by radon exposure.
Assuntos
Poluentes Radioativos do Ar/toxicidade , Células da Medula Óssea/efeitos da radiação , Perfilação da Expressão Gênica , Expressão Gênica/efeitos da radiação , Radônio/toxicidade , Administração por Inalação , Animais , Apoptose/efeitos da radiação , Células da Medula Óssea/metabolismo , Células da Medula Óssea/patologia , Regulação da Expressão Gênica/efeitos da radiação , Biblioteca Gênica , Peroxidação de Lipídeos/efeitos da radiação , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Radônio/administração & dosagem , Espécies Reativas de Oxigênio/metabolismo , Espécies Reativas de Oxigênio/efeitos da radiação , Superóxido Dismutase/metabolismoRESUMO
OBJECTIVE: To study the methylation of O-6-methylguanine-DNA methyltransferase (MGMT) and p16 gene in the sputum cells of radon-exposed population. To provide the experimental base for finding the molecular biomarker of the high risk population of the radon-induced lung cancer. METHODS: 91 radon-exposed workers were divided into 4 groups, high dosage group (> 120 WLM), middle dosage group (between 60 and 120 WLM), low dosage group (between 30 and 60 WLB) and lower dosage group (between 2 and 30 WLM) according to the accumulated exposure dosage of the radon daughters. The abnormal methylation of p16 and MGMT gene in the sputum cells of the population in the four groups was detected with the methylation specific PCR (MSP). RESULTS: There was significantly upward trend for the p16 gene methylation rate (0.00%-20.00%), the MGMT gene methylation rate (0.00%-28.00%) and the total methylation rate (0.00%-40.00%) with the increase of the accumulated exposure dosage of the radon daughters (P < 0.01). CONCLUSION: The methylation of p16 and MGMT gene is related to the accumulate exposure dosage of the radon daughters.