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@# This study sought to determine the prevalence of pathogenic and non-pathogenic bacteria in the oral cavities of children with cancer. There were 68 paediatric patients with cancer who were included in this study. Oral swab samples from the dorsum of tongues and mouth floors of these patients were subjected to culture, staining, and molecular methods to detect the bacteria. The overall prevalence of gram-positive and gram-negative bacteria was 79.4% (54/68; 95% CI = 68.4 – 87.3) and 25% (17/68; 95% CI = 16.2 – 36.4), respectively. Streptococcus salivarius and Streptococcus parasanguinis were the predominant pathogenic grampositive bacteria, while Neisseria subflava and Neisseria perflava were the most common pathogenic gram-negative bacteria. The results revealed that the number of bacteria isolates recovered in patients receiving cancer treatment was higher (55.9%) than those who had not received treatment (16.2%). Therefore, more isolated pathogenic bacteria were observed post-therapy (54.4%). Pathogenic organisms can have significant implications on patient health. Awareness of the types of bacteria inhabiting the oral cavity is essential to predict and prevent dental problems, and their associated systemic complications. Findings on the diversity of oral microflora can also provide a better understanding of the aetiology of oral diseases in paediatric patients receiving cancer treatment.
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This study aimed to investigate the mechanism of hypoxia-inducible factor-1 alpha (HIF-1α) mediated hypoxia-induced permeability changes in bladder endothelial cells. Models of in vitro hypoxic cell culture of bladder cancer, bladder cancer cells with low HIF-1α expression and HIF-1α RNA interference (RNAi) expression vector were established. Western blot and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of HIF-1α and vascular endothelial growth factor (VEGF) in each group. Bladder cell permeability was determined. Results showed that protein and mRNA expression of HIF-1α and VEGF at 3 and 12 h of hypoxia were significantly higher than normal control (P<0.05), and peaked at 12 h. HIF-1α and VEGF expression in the hypoxic group and hypoxic+3-(5′-hydroxymethyl-2′-furyl)-1-benzyl indazole (YC-1) group were significantly higher than normal control (P<0.05), while expression in the hypoxic+YC-1 group was significantly lower than the hypoxic group (P<0.05). Bladder cell permeability in the hypoxic and hypoxic+YC-1 group were significantly increased compared to normal control (P<0.05), while in the hypoxic+YC-1 group was significantly decreased compared to the hypoxic group (P<0.05). Most of the cells in the stably transfected HIF-1α RNAi expression vector pcDNA6.2-GW/EmGFP-miR-siHIF-1α expressed green fluorescence protein (GFP) under fluorescence microscope. pcDNA6.2-GW/EmGFP-miR-siHIF-1α could significantly inhibit HIF-1α gene expression (P<0.05). HIF-1α and VEGF expression in the hypoxic group and siHIF-1α hypoxic group were significantly higher than normal group (P<0.05), while expression in the siHIF-1α hypoxic group was significantly lower than the hypoxic group (P<0.05). Findings suggest that HIF-1α is an important factor in the increase of bladder cancer cell permeability.
Sujet(s)
Animaux , Rats , Tumeurs de la vessie urinaire/métabolisme , Cellules endothéliales/physiologie , Facteur de croissance endothéliale vasculaire de type A/physiologie , Sous-unité alpha du facteur-1 induit par l'hypoxie/physiologie , Hypoxie tumorale/physiologie , Perméabilité , Régulation de l'expression des gènes tumoraux/physiologie , Technique de Western , Interférence par ARN , Lignée cellulaire tumorale , Cellules endothéliales/anatomopathologie , Sous-unité alpha du facteur-1 induit par l'hypoxie/analyse , Réaction de polymérisation en chaine en temps réelRÉSUMÉ
The aim of this study was to investigate whether exogenous retinoic acid (RA) can upregulate the mRNA and protein expression of growth-associated protein 43 (GAP-43), thereby promoting brain functional recovery in a rat distal middle cerebral artery occlusion (MCAO) model of ischemia. A total of 216 male Sprague Dawley rats weighing 300–320 g were divided into 3 groups: sham-operated group, MCAO+vehicle group and MCAO+RA group. Focal cortical infarction was induced with a distal MCAO model. The expression of GAP-43 mRNA and protein in the ipsilateral perifocal region was assessed using qPCR and immunocytochemistry at 1, 3, 7, 14, 21, and 28 days after distal MCAO. In addition, an intraperitoneal injection of RA was given 12 h before MCAO and continued every day until the animal was sacrificed. Following ischemia, the expression of GAP-43 first increased considerably and then decreased. Administration of RA reduced infarction volume, promoted neurological functional recovery and upregulated expression of GAP-43. Administration of RA can ameliorate neuronal damage and promote nerve regeneration by upregulating the expression of GAP-43 in the perifocal region after distal MCAO.
Sujet(s)
Animaux , Mâle , Protéine GAP-43/métabolisme , Expression des gènes/effets des médicaments et des substances chimiques , Infarctus du territoire de l'artère cérébrale moyenne/prévention et contrôle , Neuroprotecteurs/pharmacologie , Trétinoïne/pharmacologie , Régulation positive/effets des médicaments et des substances chimiques , Encéphalopathie ischémique/prévention et contrôle , Protéine GAP-43/génétique , Immunohistochimie , Infarctus du territoire de l'artère cérébrale moyenne/métabolisme , Infarctus du territoire de l'artère cérébrale moyenne/anatomopathologie , Répartition aléatoire , Rat Sprague-Dawley , Réaction de polymérisation en chaine en temps réel , Reproductibilité des résultats , Facteurs tempsRÉSUMÉ
Different pathways act synergistically to participate in many biological processes. Thus, the purpose of our study was to extract dysregulated pathways to investigate the pathogenesis of colorectal cancer (CRC) based on the functional dependency among pathways. Protein-protein interaction (PPI) information and pathway data were retrieved from STRING and Reactome databases, respectively. After genes were aligned to the pathways, each pathway activity was calculated using the principal component analysis (PCA) method, and the seed pathway was discovered. Subsequently, we constructed the pathway interaction network (PIN), where each node represented a biological pathway based on gene expression profile, PPI data, as well as pathways. Dysregulated pathways were then selected from the PIN according to classification performance and seed pathway. A PIN including 11,960 interactions was constructed to identify dysregulated pathways. Interestingly, the interaction of mRNA splicing and mRNA splicing-major pathway had the highest score of 719.8167. Maximum change of the activity score between CRC and normal samples appeared in the pathway of DNA replication, which was selected as the seed pathway. Starting with this seed pathway, a pathway set containing 30 dysregulated pathways was obtained with an area under the curve score of 0.8598. The pathway of mRNA splicing, mRNA splicing-major pathway, and RNA polymerase I had the maximum genes of 107. Moreover, we found that these 30 pathways had crosstalks with each other. The results suggest that these dysregulated pathways might be used as biomarkers to diagnose CRC.
Sujet(s)
Humains , Adénomes/génétique , Adénomes/métabolisme , Tumeurs colorectales/génétique , Tumeurs colorectales/métabolisme , Cartes d'interactions protéiques/génétique , Aire sous la courbe , Marqueurs biologiques tumoraux/génétique , Études cas-témoins , Analyse de profil d'expression de gènes/méthodes , Régulation de l'expression des gènes , Analyse en composantes principales , Analyse par réseau de protéines , Valeurs de référence , Épissage des ARN , Transduction du signal , TranscriptomeRÉSUMÉ
Vaspin is a novel adipocytokine associated with glucose tolerance and chronic inflammation. Some studies reveal that vaspin may be involved in cardiovascular diseases. Our objective was to investigate the relationship between serum vaspin levels and endothelial function in patients with ankylosing spondylitis. One hundred and twenty patients with newly diagnosed ankylosing spondylitis and 100 healthy subjects were studied. Serum vaspin levels were measured with enzyme-linked immunosorbent assay. High resolution ultrasound was used to measure brachial artery diameter at rest, after reactive hyperemia (flow-mediated dilation, FMD) and after sublingual glyceryltrinitrate. Serum vaspin level in patients was 1.92±1.03 ng/mL, which was significantly lower than that in healthy subjects (2.88±0.81 ng/mL). By dividing the distribution of serum vaspin levels into quartiles, FMD levels increased gradually with the increase of serum vaspin levels in patients (P<0.01). Univariate analysis showed a correlation between vaspin and FMD (r=0.73, P=0.003), low-density lipoprotein cholesterol (r=-0.45, P=0.033), high-density lipoprotein cholesterol (r=0.63, P=0.025), fasting blood glucose (r=-0.79, P=0.006), triglycerides (TG) (r=-0.68, P=0.036), systolic blood pressure (r=-0.35, P=0.021), C-reactive protein (r=-0.67, P=0.011), homeostatic model assessment of insulin resistance (HOMA-IR) (r=-0.77, P=0.023) and erythrocyte sedimentation rate (r=-0.88, P=0.039) in patients. Multivariate analysis indicated that serum vaspin levels were independently associated with FMD, HOMA-IR and TG in patients. Our study found that serum vaspin levels were decreased in patients with ankylosing spondylitis and were associated with FMD levels. Vaspin may serve as an independent marker for detecting early stage atherosclerosis in patients with ankylosing spondylitis.
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Humains , Mâle , Femelle , Adulte , Adulte d'âge moyen , Sujet âgé , Pelvispondylite rhumatismale/physiopathologie , Pelvispondylite rhumatismale/sang , Endothélium vasculaire/physiopathologie , Serpines/sang , Valeurs de référence , Triglycéride/sang , Glycémie/analyse , Artère brachiale/anatomopathologie , Artère brachiale/imagerie diagnostique , Insulinorésistance , Marqueurs biologiques/sang , Études cas-témoins , Modèles linéaires , Cholestérol/sang , Facteurs de risque , Analyse de varianceRÉSUMÉ
Pancreatic ductal adenocarcinoma (PDAC) has a poor 5-year survival rate of 5%. Biomarkers for the early detection of pancreatic cancer are urgently needed. Transforming growth factor-beta1 (TGF-β1) is elevated in the tissues and plasma of patients with PDAC. However, no studies systemically report prognostic significance of plasma TGF-β1 levels in PDAC. In the present study, we assessed the prognostic significance of serum TGF-β levels in patients with PDAC. TGF-β levels were determined in serum from 146 PDAC patients, and 58 patients with benign pancreatic conditions. Regression models were used to correlate TGF-β levels to gender, age, stage, class, and metastasis. Survival analyses were performed using multivariate Cox models. Serum levels of TGF-β1 distinguished PDAC from benign pancreatic conditions (P<0.001) and healthy control subjects (P<0.001). Serum levels of TGF-β also distinguished tumor stage (P=0.002) and lymph node metastasis (P=0.001). High serum levels of TGF-β1 were significantly correlated with reduced patient survival. Multivariate analysis revealed that TGF-β1, lymph node metastasis and tumor stage were independent factors for PDAC survival. Our results indicate that serum TGF-β1 may be used as a potential prognostic marker for PDAC.
Sujet(s)
Humains , Tumeurs du pancréas/sang , Marqueurs biologiques tumoraux/sang , Carcinome du canal pancréatique/sang , Facteur de croissance transformant bêta-1/sang , Tumeurs du pancréas/diagnostic , Tumeurs du pancréas/secondaire , Pronostic , Études rétrospectives , Sensibilité et spécificité , Carcinome du canal pancréatique/diagnostic , Estimation de Kaplan-MeierRÉSUMÉ
Purpose: To establish a new genotyping method for Vibrio cholerae and compare it with other methods. Materials and Methods: In the current study, a modified inter simple sequence repeat-polymerase chain reaction (MISSR-PCR) system was developed via several rounds of optimisation. Comparison study was then conducted between MISSR-PCR and three other methods, including enterobacterial repetitive intergenic consensus sequences-based PCR (ERIC-PCR), randomly amplified polymorphic DNA (RAPD) and 16S rRNA evolutionary clock, for the detection and genetic tracing of Vibrio cholerae isolated from seafood in China. Result: The results indicated that the MISSRPCR system could generate the highest polymorphic fingerprinting map in a single round PCR and showed the best discriminatory ability for Vibrio cholerae genotyping by clearly separating toxigenic/nontoxigenic strains, local/foreign strains, and O1/O139/non-O1/non-O139 serogroup strains, comparing to ERIC-PCR, RAPD and 16S rRNA evolutionary clock. Moreover, the MISSR-PCR is superior to previously described traditional simple sequence repeat based PCR method on genotyping by more clearly separating different clusters. Conclusion: To the best of our knowledge, this is the first head-to-head comparison of four detection and genotyping methods for Vibrio cholerae. The MISSRPCR system established here could serve as a simple, quick, reliable and cost-effective tool for the genotyping and epidemiological study.
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As an essential trace element, copper can be toxic in mammalian cells when present in excess. Metallothioneins (MTs) are small, cysteine-rich proteins that avidly bind copper and thus play an important role in detoxification. Yeast CUP1 is a member of the MT gene family. The aim of this study was to determine whether yeast CUP1 could bind copper effectively and protect cells against copper stress. In this study, CUP1 expression was determined by quantitative real-time PCR, and copper content was detected by inductively coupled plasma mass spectrometry. Production of intracellular reactive oxygen species (ROS) was evaluated using the 2',7'-dichlorofluorescein-diacetate (DCFH-DA) assay. Cellular viability was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and the cell cycle distribution of CUP1 was analyzed by fluorescence-activated cell sorting. The data indicated that overexpression of yeast CUP1 in HeLa cells played a protective role against copper-induced stress, leading to increased cellular viability (P<0.05) and decreased ROS production (P<0.05). It was also observed that overexpression of yeast CUP1 reduced the percentage of G1 cells and increased the percentage of S cells, which suggested that it contributed to cell viability. We found that overexpression of yeast CUP1 protected HeLa cells against copper stress. These results offer useful data to elucidate the mechanism of the MT gene on copper metabolism in mammalian cells.
Sujet(s)
Animaux , Humains , Mammifères/physiologie , Phéromones/physiologie , Comportement animal/physiologie , Comportement/physiologie , Odorisants , Bulbe olfactif/physiologie , Muqueuse olfactive/physiologie , Voies olfactives/anatomie et histologie , Voies olfactives/physiologie , Neurorécepteurs olfactifs/physiologie , Phéromones humaines/physiologie , Odorat/physiologieRÉSUMÉ
We evaluated the effect of puerarin on spatial learning and memory ability of mice with chronic alcohol poisoning. A total of 30 male C57BL/6 mice were randomly divided into model, puerarin, and control groups (n=10 each). The model group received 60% (v/v) ethanol by intragastric administration followed by intraperitoneal injection of normal saline 30 min later. The puerarin group received intragastric 60% ethanol followed by intraperitoneal puerarin 30 min later, and the control group received intragastric saline followed by intraperitoneal saline. Six weeks after treatment, the Morris water maze and Tru Scan behavioral tests and immunofluorescence staining of cerebral cortex and hippocampal neurons (by Neu-N) and microglia (by Ib1) were conducted. Glutamic acid (Glu) and gamma amino butyric acid (GABA) in the cortex and hippocampus were assayed by high-performance liquid chromatography (HPLC), and tumor necrosis factor (TNF)-α and interleukin (IL)-1β were determined by ELISA. Compared with mice in the control group, escape latency and distance were prolonged, and spontaneous movement distance was shortened (P<0.05) by puerarin. The number of microglia was increased in both the cortex and hippocampal dentate gyrus (P<0.01), and neurons were reduced only in the hippocampal dentate gyrus (P<0.01) in puerarin-treated mice. In the model group, Glu and GABA levels decreased (P<0.05), and Glu/GABA, TNF-α, and IL-1β increased (P<0.01) with puerarin treatment, returning to near normal levels. In conclusion, puerarin protected against the effects of chronic alcohol poisoning on spatial learning and memory ability primarily because of anti-inflammatory activity and regulation of the balance of Glu and GABA.
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Animaux , Mâle , Éthanol/intoxication , Isoflavones/usage thérapeutique , Apprentissage du labyrinthe/effets des médicaments et des substances chimiques , Troubles de la mémoire/prévention et contrôle , Neuroprotecteurs/usage thérapeutique , Mémoire spatiale/effets des médicaments et des substances chimiques , Vasodilatateurs/usage thérapeutique , Alcoolisme/complications , Chromatographie en phase liquide à haute performance , Cortex cérébral/composition chimique , Cortex cérébral/effets des médicaments et des substances chimiques , Test ELISA , Acide glutamique/analyse , Interleukine-1 bêta/analyse , Isoflavones/pharmacologie , Troubles de la mémoire/induit chimiquement , Troubles de la mémoire/traitement médicamenteux , Microglie/effets des médicaments et des substances chimiques , Neuroprotecteurs/pharmacologie , Répartition aléatoire , Facteurs temps , Résultat thérapeutique , Facteur de nécrose tumorale alpha/analyse , Vasodilatateurs/pharmacologie , Acide gamma-amino-butyrique/analyseRÉSUMÉ
The purpose of this study was to explore cytokine expression patterns and cytogenetic abnormalities of mesenchymal stem cells (MSCs) from the bone marrow microenvironment of Chinese patients with myelodysplastic syndromes (MDS). Bone marrow samples were obtained from 30 cases of MDS (MDS group) and 30 healthy donors (control group). The expression pattern of cytokines was detected by customized protein array. The karyotypes of MSCs were analyzed using fluorescence in situ hybridization. Compared with the control group, leukemia inhibitory factor, stem cell factor (SCF), stromal cell-derived factor (SDF-1), bone morphogenetic protein 4, hematopoietic stem cell (HSC) stimulating factor, and transforming growth factor-β in the MDS group were significantly downregulated (P<0.05), while interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and programmed death ligand (B7-H1) were significantly upregulated (P<0.05). For chromosome abnormality analysis, the detection rate of abnormal karyotypes (+8, -8, -20, 20q-, -Y, -7, 5q-) was 30% in the MDS group and 0% in the control group. In conclusion, the up- and downregulated expression of these cytokines might play a key role in the pathogenesis of MDS. Among them, SCF and SDF-1 may play roles in the apoptosis of HSCs in MDS; and IFN-γ, TNF-α, and B7-H1 may be associated with apoptosis of bone marrow cells in MDS. In addition, the abnormal karyotypes might be actively involved in the pathogenesis of MDS. Further studies are required to determine the role of abnormal karyotypes in the occurrence and development of MDS.
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Sujet âgé , Humains , Mâle , Os et tissu osseux/composition chimique , Plomb/analyse , Professions , Exposition professionnelle/effets indésirables , Anciens combattants , Os et tissu osseux , Études longitudinales , Enquêtes et questionnaires , États-UnisRÉSUMÉ
Liver cirrhosis is one of the most common diseases of Chinese patients. Herein, we report the high expression of a newly identified histone 3 lysine 4 demethylase, retinoblastoma binding protein 2 (RBP2), and its role in liver cirrhosis in humans. The siRNA knockdown of RBP2 expression in hepatic stellate cells (HSCs) reduced levels of α-smooth muscle actin (α-SMA) and vimentin and decreased the proliferation of HSCs; and overexpression of RBP2 increased α-SMA and vimentin levels. Treatment with transforming growth factor β (TGF-β) upregulated the expression of RBP2, α-SMA, and vimentin, and the siRNA knockdown of RBP2 expression attenuated TGF-β-mediated upregulation of α-SMA and vimentin expression and HSC proliferation. Furthermore, RBP2 was highly expressed in cirrhotic rat livers. Therefore, RBP2 may participate in the pathogenesis of liver cirrhosis by regulating the expression of α-SMA and vimentin. RBP2 may be a useful marker for the diagnosis and treatment of liver cirrhosis.
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Animaux , Humains , Mâle , Actines/métabolisme , Cellules étoilées du foie/métabolisme , Histone Demethylases/métabolisme , Cirrhose du foie/métabolisme , /métabolisme , Vimentine/métabolisme , Technique de Western , Prolifération cellulaire , Modèles animaux de maladie humaine , Expression des gènes , Techniques de knock-down de gènes , Rat Wistar , Petit ARN interférent/métabolisme , Facteur de croissance transformant bêta/métabolismeRÉSUMÉ
OBJECTIVE: We attempted to evaluate maternal thyroid function in a new self-sequential longitudinal reference interval (SLRI) which we established recently. By this method, we analysed the correlation between pregnancy outcome, neonatal thyroid stimulating hormone (TSH) level and maternal thyroid diseases. METHODS: A total of 1744 pregnant women participated in the study and 1747 babies were born from those women (three bore twins). The levels of TSH, free thyroxine (FT4) and thyroid peroxidase antibodies (TPO-Ab) of mothers were quantified by electrochemistry immunoassay (ECL). The levels of neonatal blood TSH were detected by time-resolved fluorescence immunoassay (TRFIA). All data were collected and statistically analysed by SPSS 13.0 software. RESULTS: With our new SLRI method, we found that 0.11%~3.84% pregnant women would get thyroid diseases. Subclinical hypothyroidism was the most common maternal thyroid disorder. Being positive for thyroid peroxidase antibodies was a significant risk factor of subclinical hypothyroidism during pregnancy. The median, P2.5~P97.5, and interquartile range (IQR) of neonatal TSH (N-TSH) of 1747 babies were 2.72 mIU/L, 0.10~8.01 mIU/L and 2.62 mIU/L, respectively; 28.6% of pregnant women with thyroid diseases developed pregnancy complications. The prevalence was significantly higher than in the normal thyroid function group (p< 0.001). The levels of N-TSH were low correlated with maternal TSH levels (p < 0.05), but there were no significant correlations between N-TSH and maternal FT4 and maternal TPO-Ab (p > 0.05). CONCLUSIONS: Thyroid disorders, especially subclinical hypothyroidism, are common in pregnant women. These disorders are associated with pregnancy and fetal outcome. Routine maternal thyroid function screening is important and should be recommended.
OBJETIVO: Intentamos evaluar la función tiroidea materna en un nuevo intervalo de referencia longitudinal auto-secuencial (SLRI) que establecimos recientemente. Por este método, analizamos la correlación entre el resultado del embarazo, el nivel de la hormona estimulante de la tiroides (TSH) en neonatos, y las enfermedades tiroideas maternas MÉTODOS: Un total de 1744 mujeres embarazadas participó en el estudio y 1747 bebés nacieron de esas mujeres (tres de ellas tuvieron gemelos). Los niveles de TSH, la tiroxina libre (FT4), y los anticuerpos de la peroxidasa tiroidea (TPO-Ab) de las madres, fueron cuantificados mediante inmunoensayo electroquímico (ECL). Los niveles de TSH en la sangre de los neonatos, fueron determinados mediante inmunoensayo por fluorescencia resuelto en el tiempo (TRFIA). Todos los datos fueron recogidos y analizados estadísticamente usando el software SPSS 13.0 RESULTADOS: Con nuestro nuevo método SLRI, encontramos que 0.11%~3.84% de las mujeres embarazadas contraerán enfermedades tiroideas. El hipotiroidismo subclínico fue el trastorno de la tiroides materna más común. Ser positivo a los anticuerpos de la peroxidasa tiroidea fue un factor de riesgo significativo del hipotiroidismo subclínico durante el embarazo. La mediana, P2.5~P97.5, y el rango intercuartil (IQR) de la TSH (N-TSH) neonatal de los 1747 bebés fueron 2.72 mIU/L, 0.10~8.01 mIU/L y 2.62 mIU/L respectivamente. El 28.6% de las mujeres embarazadas que tenían enfermedades tiroideas, desarrollaron complicaciones del embarazo. La prevalencia fue significativamente más alta que en el grupo con función tiroidea normal (p < 0.001). Los niveles de N-TSH fueron bajos en correlación con los niveles de TSH maternos (p < 0.05), pero no hubo ninguna correlación significativa entre la N-TSH y la FT4 materna, y la TPO-Ab materna (p > 0.05). CONCLUSIÓNS: Los trastornos tiroideos, especialmente el hipotiroidismo, son comunes en las mujeres embarazadas.Estos trastornos se hallan asociados con el resultado del embarazo y el resultado fetal. El tamizaje de rutina de la función tiroidea materna es importante y debe recomendarse.
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Humains , Femelle , Nouveau-né , Adulte , Peroxidases/sang , Complications de la grossesse/diagnostic , Maladies de la thyroïde/diagnostic , Thyréostimuline/sang , Complications de la grossesse/sang , Complications de la grossesse/épidémiologie , Trimestres de grossesse/sang , Valeurs de référence , Maladies de la thyroïde/sang , Maladies de la thyroïde/épidémiologie , Tests de la fonction thyroïdienne/méthodes , Issue de la grossesse , Chine/épidémiologie , Dépistage néonatalRÉSUMÉ
The microenvironment of the tumor plays an important role in facilitating cancer progression and activating dormant cancer cells. Most tumors are infiltrated with inflammatory cells which secrete cytokines such as tumor necrosis factor-alpha (TNF-alpha). To evaluate the role of TNF-alpha in the development of cancer we studied its effects on cell migration with a migration assay. The migrating cell number in TNF-alpha-treated group is about 2-fold of that of the control group. Accordingly, the expression of E-cadherin was decreased and the expression of vimentin was increased upon TNF-alpha treatment. These results showed that TNF-alpha can promote epithelial-mesenchymal transition (EMT) of MCF-7 cells. Further, we found that the expression of Snail, an important transcription factor in EMT, was increased in this process, which is inhibited by the nuclear factor kappa B (NFkB) inhibitor aspirin while not affected by the reactive oxygen species (ROS) scavenger N-acetyl cysteine. Consistently, specific inhibition of NFkB by the mutant IkBalpha also blocked the TNF-alpha-induced upregulation of Snail promoter activity. Thus, the activation of NFkB, which causes an increase in the expression of the transcription factor Snail is essential in the TNF-alpha-induced EMT. ROS caused by TNF-alpha seemed to play a minor role in the TNF-alpha-induced EMT of MCF-7 cells, though ROS per se can promote EMT. These findings suggest that different mechanisms might be responsible for TNF-alpha - and ROS-induced EMT, indicating the need for different strategies for the prevention of tumor metastasis induced by different stimuli.
Sujet(s)
Humains , Cellules épithéliales/métabolisme , Mésoderme/cytologie , Facteur de transcription NF-kappa B/physiologie , Espèces réactives de l'oxygène/métabolisme , Facteurs de transcription/métabolisme , Facteur de nécrose tumorale alpha/pharmacologie , Technique de Western , Études cas-témoins , Lignée cellulaire tumorale , Cadhérines/métabolisme , Mouvement cellulaire/effets des médicaments et des substances chimiques , Cellules épithéliales/anatomopathologie , Mésoderme/métabolisme , Facteur de transcription NF-kappa B/métabolisme , RT-PCR , Facteur de nécrose tumorale alpha/physiologie , Vimentine/métabolismeRÉSUMÉ
Establishment of a cell-mediated gene transfer system has potential as a new breeding technique for commercially valuable fishes. As an important step toward developing an inter-species chimera, cells from blastula-stage embryos of rosy barb (Puntius conchonius) were transplanted into zebrafish (Danio rerio) blastula-stage embryos to observe the development of the recipient. From the total of 473 transplants obtained only a fraction of 13 chimeras appeared perfectly normal after one month. Over two in normal 13 chimeras showed some characters from the donor cells with scarce pigmentation. This is the first successful inter-species study on zebrafish by using blastula cell transplants from rosy barb both belonging to the same family cyprinidae.