RESUMEN
PURPOSE: To investigate the effect of microRNA-543 (miR-543) on the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of triple-negative breast cancer (TNBC) cells, and the associated mechanism. METHODS: Human breast cancer cells (MDA-MB-231, HCC1937, and MCF-7, ZR-75-1) and normal human breast epithelial cell line (MCF10A) were transfected with miR-543 mimics or inhibitor using lipofectamine 2000. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blotting were used to determine the mRNA and protein expression levels of miR-543, actin-like protein 6A (ACTL6A), vimentin, Snail, and E-cadherin in breast cancer cells/tissue. Cell counting kit-8 (CCK-8), wound-healing, and Transwell assays were used to measure the effect of miR-543 on TNBC cell proliferation, invasion, and migration. Overall survival was determined using data from Gene Expression Omnibus (GEO) and Cancer Genome Atlas (TCGA) databases. Bioinformatics analysis and luciferase reporter gene assay were used to determine the regulatory effect of miR-543 on ACTL6A. RESULTS: The level of expression of miR-543 was significantly lower in breast cancer cells/tissue than in normal human breast epithelial cell/tissue (p < 0.05). MicroRNA-543 expression level was significantly reduced in TNBC cells/tissue, relative to the other breast cancer cells/normal breast tissue (p < 0.05). MicroRNA-543 significantly suppressed tumor growth and the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of TNBC cells, in mouse xenograft model (p < 0.05). CONCLUSIONS: miR-543 influences the biological behavior of TNBC cells by directly targeting ACTL6A gene. miR-543 could serve as a novel diagnostic and therapeutic target for TNBC.
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Actinas/fisiología , Movimiento Celular , Proliferación Celular , Proteínas Cromosómicas no Histona/fisiología , Proteínas de Unión al ADN/fisiología , Regulación hacia Abajo , Transición Epitelial-Mesenquimal , MicroARNs/fisiología , Neoplasias de la Mama Triple Negativas/patología , Animales , Humanos , Ratones , Invasividad Neoplásica , Células Tumorales CultivadasRESUMEN
Lipid metabolism dysfunction is closely related to obesity, inflammation, diabetes, lipodystrophy, cardiovascular disease. Along with having a positive effect on energy homeostasis during fasting or prolonged exercise through mitochondrial fatty acid oxidation (FAO), more than two dozen enzymes and transport proteins are involved in the activation and transport of fatty acids into the mitochondrial, providing insights into their critical roles in metabolism. CPT1A has been reported to be expressed ubiquitously in the body and associated with dire consequences affecting fat deposition as the key rate-limiting enzyme of FAO. However, there is a dearth of data on the specific role of CPT1A on adipogenic differentiation and adipocyte lipolysis on chicken. This study assessed CPT1A's function in adipocyte differentiation andadipocyte lipolysis, and the mechanisms were investigated. We found that CPT1A knockdown (KD) promotes the differentiation of chicken preadipocytes into mature adipocytes. CPT1A KD increased PPARγ protein expression level. Expression levels of lipid synthesis-related genes were increased, and lipolysis genes were reduced. Also, CPT1A KD can encourage the formation of lipid droplets. So our results confirmed that knockdown of CPT1A induced the lipid differentiation and inhibited the ß-oxidation process to promote the formation of lipid droplets. These findings may deepen our understanding on CPT1A function, especially its regulatory role in adipocyte biology.(AU)
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Animales , Carnitina O-Palmitoiltransferasa , Pollos/fisiología , Adipocitos/clasificación , Metabolismo de los Lípidos , Técnicas de Silenciamiento del Gen/instrumentaciónRESUMEN
Intramuscular fat (IMF) content is a crucial parameter for estimating meat quality. Growing evidence indicates that gene regulation plays an important role in IMF deposition. This study aimed to determine the function of Mfsd2a in chicken intramuscular preadipocytes. In the present study, high Mfsd2a mRNA levels were observed in the liver and adipose tissues of broilers. Subsequently, we synthesized small interfering RNAs to silence the expression of Mfsd2a in chicken intramuscular preadipocytes. The following results suggested that CDK2, PCNA, CCND1, CCND2 and MKI67 were inhibited, with CCK-8 and EdU assays revealing that cell proliferation was inhibited. Scratch test showed that cell migration ratios were declined. We also found that Mfsd2a silencing decreased the mRNA levels of PPARγ, RXRG and their target genes. The similar results were found in some key genes that contribute to lipid synthesis, including C/EBPα, C/EBPß, FABP4, FASN, ACACA and ACSL1. Finally, Oil red O staining showed that IMF accumulation was blocked after Mfsd2a silencing. In conclusion, our results implied that Mfsd2a promotes the proliferation and migration of chicken intramuscular preadipocytes, as well as the differentiation and adipogenesis through PPARγ signaling pathway, which may provide a potential target to improve chicken meat quality.(AU)
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Animales , Pollos , Antígeno Nuclear de Célula en Proliferación , Adipogénesis , SimportadoresRESUMEN
Utilizando um anticorpo monoclonal contra a aflatoxina B1 (AFB1) como ligante, foi identificado um mimotopo específico de aflatoxina B1 após se realizarem quatro ciclos de seleção biológica de 7-peptídeos aleatórios em biblioteca de fago exibida. O mimotopo é denominado P10, e sua sequência de aminoácidos é YRRHEKD. O soro imunológico de ratos Balb/c imunizados com P10 foi especificamente ligado à aflatoxina B1-albumina, indicando que o anticorpo era específico ao AFB1. Esses resultados sugerem que é possível desenvolver a vacina baseada em mimotopo associado à toxina.(AU)
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Animales , Ratas , Vacunas Fúngicas/análisis , Aflatoxina B1 , Aptámeros de Péptidos/inmunología , Inmunogenicidad Vacunal , Ratones Endogámicos BALB C/inmunologíaRESUMEN
Utilizando um anticorpo monoclonal contra a aflatoxina B1 (AFB1) como ligante, foi identificado um mimotopo específico de aflatoxina B1 após se realizarem quatro ciclos de seleção biológica de 7-peptídeos aleatórios em biblioteca de fago exibida. O mimotopo é denominado P10, e sua sequência de aminoácidos é YRRHEKD. O soro imunológico de ratos Balb/c imunizados com P10 foi especificamente ligado à aflatoxina B1-albumina, indicando que o anticorpo era específico ao AFB1. Esses resultados sugerem que é possível desenvolver a vacina baseada em mimotopo associado à toxina.(AU)
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Animales , Ratas , Vacunas Fúngicas/análisis , Aflatoxina B1 , Aptámeros de Péptidos/inmunología , Inmunogenicidad Vacunal , Ratones Endogámicos BALB C/inmunologíaRESUMEN
BACKGROUND: Angiogenesis is an indispensable step in the growth and invasiveness of breast cancers involving a series of exquisite molecular steps. Pro-angiogenic factors, including vascular endothelial growth factor (VEGF), have been recognized as pivotal therapeutic targets in the treatment of breast cancer. More recently, a highly conserved transcription factor Twist has been reported to be involved in tumor angiogenesis and metastasis. METHODS: The expression of VEGF-C and Twist was immunohistochemically determined in tissue samples of primary tumors from 408 patients undergoing curative surgical resection for breast cancer. The correlations of VEGF-C and Twist expressions with clinicopathologic parameters as well as survival outcomes were evaluated. RESULTS: Of the 408 patients evaluated, approximately 70% had high expression of VEGF-C which was significantly associated with advanced tumor stages (P = 0.019). Similarly, VEGF-C expression was associated with the proliferation index Ki67, N3 lymph node metastasis, and D2-40-positive lymphatic vessel invasion (LVI) in a univariate analysis. Furthermore, patients with high expressions of VEGF-C and Twist (V + T+) had significantly increased lymph node metastasis, higher clinical stage, and worse disease-free survival, DFS (P = 0.001) and overall survival, OS (P = 0.011). CONCLUSIONS: Our results suggested that co-expression of VEGF-C and Twist was associated with larger tumor size, higher numbers of lymph node involvement, D2-40-positive LVI, higher risk of distant metastasis, and worse DFS or OS in breast cancer patients.
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Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/patología , Carcinoma Ductal de Mama/patología , Proteínas Nucleares/metabolismo , Proteína 1 Relacionada con Twist/metabolismo , Factor C de Crecimiento Endotelial Vascular/metabolismo , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/cirugía , Carcinoma Ductal de Mama/metabolismo , Carcinoma Ductal de Mama/cirugía , Femenino , Estudios de Seguimiento , Regulación Neoplásica de la Expresión Génica , Humanos , Persona de Mediana Edad , Pronóstico , Tasa de SupervivenciaRESUMEN
PURPOSE: Glioma is a common malignant tumor of the central nervous system, which is characterized by a low cure rate, high morbidity, and high recurrence rate. Consequently, it is imperative to explore some indicators for prognostic prediction in glioma. METHODS: We obtained glioma data from The Cancer Genome Atlas (TCGA). Differentially expressed genes (DEGs) were obtained by R software from TCGA data sets. Through Cox regression analysis, risk scores were obtained to assess the weighted gene-expression levels, which could predict the prognosis of patients with glioma. The validity and the prognostic value of this model in glioma were confirmed by the manifestation of receiver-operating characteristic (ROC) curves, area under the curve (AUC), and 5-year overall survival (OS). RESULTS: In total, 920 DEGs of transcriptome genes in glioma were extracted from the TCGA database. We identified a novel seven-gene signature associated with glioma. Among them, AL118505.1 and SMOC1 were positively related to the 5-year OS of patients with glioma, showing a better prognosis for glioma; however, RAB42, SHOX2, IGFBP2, HIST1H3G, and IGF2BP3 were negatively related to 5-year OS, displaying a worse prognosis. In addition, according to risk scores, AL118505.1 was also a protective factor, while others were risk factors. Furthermore, the expression levels of SHOX2, IGFBP2, and IGF2BP3 were significantly positively correlated with glioma grades. Receiver-operating characteristic (ROC) curve assessed the accuracy and sensitivity of the gene signature. Each of the seven genes for patients with the distribution of the risk score was presented in the heat map. CONCLUSION: We identified a novel seven-gene signature in patients with glioma, which could be used as a predictor for the prognosis of patients with glioma in the future.
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Neoplasias Encefálicas/genética , Expresión Génica , Genes Relacionados con las Neoplasias , Glioma/genética , Área Bajo la Curva , Neoplasias Encefálicas/mortalidad , Glioma/mortalidad , Proteínas de Homeodominio/genética , Humanos , Proteína 2 de Unión a Factor de Crecimiento Similar a la Insulina/genética , Proteínas de Neoplasias/genética , Osteonectina/genética , Pronóstico , Proteínas de Unión al ARN/genética , Curva ROC , Análisis de Regresión , Riesgo , TranscriptomaRESUMEN
OBJECTIVES: The aim of this study was to investigate the effects of primary healthcare decentralization on type 2 diabetes mellitus mortality and morbidity in different municipalities of a developing country. STUDY DESIGN: This was a retrospective study based on a panel of annual data from 5560 Brazilian municipalities from 2000 to 2011. METHODS: The investigation used the staggered municipal adoption of a federal health information program as a quasi-experiment to identify the treatment effects of health decentralization on diabetes indicators. Using Difference-in-Differences models and instrumental variables, we analyzed the effects of primary healthcare decentralization on diabetes rates (i.e. diabetes deaths and hospitalizations by the number of people with a diabetes diagnosis and by population). RESULTS: Evidence suggests improvements in universal access to primary health care and progress in the average health outcomes related to diabetes mortality (reduction of 30%) and hospitalization (reduction of 2.3%) due to decentralization. Effects are further pronounced in developed regions with higher incomes, while the poorest and less developed regions showed virtually no effect. CONCLUSIONS: These results demonstrate that there are particular preconditions for successful primary health decentralization, especially related to returns of scale (big health facilities are associated with low cost per treatment), lack of human and physical capital, and government coordination problems.
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Diabetes Mellitus Tipo 2/epidemiología , Diabetes Mellitus Tipo 2/terapia , Política , Atención Primaria de Salud/organización & administración , Brasil/epidemiología , Ciudades , Diabetes Mellitus Tipo 2/mortalidad , Accesibilidad a los Servicios de Salud , Hospitalización/estadística & datos numéricos , Humanos , Estudios Retrospectivos , Factores SocioeconómicosRESUMEN
Growth traits are important economic traits in broiler chicken production. AluI and Hin1I loci are two restriction sites, which are respectively located in exons 2 and 3 of the IGF-1R gene. These two loci are significantly related to the growth traits in Jinghai Yellow chickens. In this study, a correlation analysis was performed between these two loci and the growth traits of Bian chickens. The results showed a G376A mutation at the AluI site and a C919A mutation at the Hin1I site, which respectively resulted in three genotypes AA, AB, and BB in exon 2 and three genotypes CC, CD, and DD in exon 3. Correlation analysis showed that the female Bian chickens with the AA genotype of the AluI locus had higher body weights than those with the AB genotype (P < 0.05) at 8, 14, 16, and 18 weeks; individuals with CD genotype of Hin1I locus had higher body weights at 6, 8, 10, 12, and 14 weeks compared to the CC genotype (P < 0.05 or P < 0.01). Combined genotypes analysis showed that at the age of 8, 14, 16, and 18 weeks, the body weight of AACC genotype combination was higher than that of the ABCC genotype combination (P < 0.05); at the age of 6, 8, 12, 14, 16, and 18 weeks, the AACD genotype combination had higher (P < 0.05 or P < 0.01) body weight than that of the ABCC genotype.
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Peso Corporal/genética , Pollos/genética , Polimorfismo de Longitud del Fragmento de Restricción , Carácter Cuantitativo Heredable , Receptores de Somatomedina/genética , Animales , Pollos/crecimiento & desarrollo , GenotipoRESUMEN
Reproductive traits have long been studied and have an important influence on chicken breeding. To identify quantitative trait loci affecting reproductive traits, a genome-wide analysis of a Chinese chicken breed was performed to analyze age at first egg body weight at first egg, first egg weight, egg weight at the age of 300 days, egg weight at the age of 462 days, egg number at the age of 300 days, egg number between the ages of 300 and 462 days and egg number at the age of 462 days. Nineteen SNPs related to reproductive traits were presented (P < 1.80E-6). Nine of the 19 SNPs had a significant effect on BWF, six SNPs were significantly associated with egg weight, and four SNPs were significantly associated with egg number. These SNPs were located near to or in 17 genes including FAM184B, HTT, KCNH7, CDC42BPA, KCNIP4, GJA5, CBFB, and GPC6. The present results may be beneficial for reproductive research and may be used in marker-assisted selection in future studies. These results could potentially benefit further breeding programs, especially in Jinghai Yellow Chicken.
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Pollos/genética , Reproducción/genética , Animales , Biomarcadores , China , Femenino , Estudio de Asociación del Genoma Completo , Haplotipos , Desequilibrio de Ligamiento , Masculino , Polimorfismo de Nucleótido Simple , Sitios de Carácter Cuantitativo , Carácter Cuantitativo Heredable , Selección ArtificialRESUMEN
Hurood cheese (HC) and Jueke (Jk) are 2 traditional fermented dairy products produced from raw milk (RM) in the Inner Mongolia region of China. They have a long history of production and consumption. The microbial compositions of RM, HC, and Jk vary greatly, and are influenced by their geographical origins and unique processing methods. In this study, 2 batches of RM, HC, and Jk samples were collected (April and August 2015) from the Zhenglan Banner, a region located in the southern part of Inner Mongolian belonging to the Xilingol league prefecture. The bacterial and fungal diversities of the samples were determined by 16S rRNA and 18S rRNA gene sequence analysis, respectively. A total of 112 bacterial and 30 fungal sequences were identified, with Firmicutes and Ascomycota being the predominant phyla for bacteria and fungi, respectively. Lactococcus and Lactobacillus were identified as the main bacterial genera, whereas Kluyveromyces was the predominant fungus identified in the 3 dairy products. Different bacterial and fungal compositions were observed in RM, HC, and Jk samples collected at different times. These results suggested that time of production may be an important factor influencing the microbial diversity present in RM, HC, and Jk.
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Productos Lácteos Cultivados/microbiología , ADN Bacteriano/genética , Leche/microbiología , Animales , Bacterias/genética , Bacterias/aislamiento & purificación , Biodiversidad , Queso/microbiología , China , Microbiología de Alimentos , Hongos/genética , Hongos/aislamiento & purificación , Leche/química , ARN Ribosómico 16S/genética , ARN Ribosómico 18S/genética , Análisis de Secuencia de ADNRESUMEN
The insulin-like growth factor, IGF-I, plays an important role in the development of growth and reproductive traits. Single-strand conformation polymorphism analysis was used to detect and analyze polymorphisms and expression profiles of the IGF-I gene and its association with growth and reproductive traits of Jinghai yellow chickens. A point mutation g.295T>C was detected in the IGF-I gene with three genotypes CC, CT, and TT. The CT and TT genotypes were found to be significantly (P < 0.05) superior to the CC genotype in 8-, 10-, 12-, 14-, and 16-week-age weight of chickens (for growth traits) and in body weight at first egg-laying (for reproductive traits). Comparison of the expression level between males and females showed similarity in their expression curves, with females showing relatively higher IGF-I expression than males in all studied tissues. A similar IGF-I expression pattern was observed in the breast and leg muscles of both males and females, with the leg muscle showing relative higher IGF-I expression than the breast muscle. Our results indicate that, g.295T>C mutation in the IGF-I gene affects certain growth and reproductive traits, and it could be used to provide a theoretical basis as well as marker-assisted selection to upgrade the development of Jinghai yellow chickens in future.
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Proteínas Aviares/genética , Pollos/crecimiento & desarrollo , Factor I del Crecimiento Similar a la Insulina/genética , Reproducción , Animales , Pollos/genética , Femenino , Estudios de Asociación Genética , Genotipo , Fenotipo , Polimorfismo Conformacional Retorcido-Simple , Sitios de Carácter Cuantitativo , Selección GenéticaRESUMEN
Germline stem cells are the only such capable of transmitting genetic information in vivo. The isolation and culture of these cells in vitro provide a unique model to understand sperm differentiation and hence, spermatogenesis and male fertility. In this study, we isolated, purified, and cultured germline stem cells from the testes of newborn calves. Moreover, we investigated the effects of glial cell line-derived neurotrophic factor (GDNF) and leukemia-inhibitory factor (LIF) on their proliferation. Male calf germline stem cells were found to be pluripotent, and able to form grape-like and embryonic stem cell (ES)-like colonies when cultured. GDNF promoted proliferation of the former, whereas LIF induced growth of the latter. The grape-like colonies retained their germline stem cell characteristics, whereas the ES-like colonies demonstrated more primitive attributes. This investigation established a male calf germline stem cell culture model that may serve as a foundation for further studies aiming to understand the properties of such cells.
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Células Madre Embrionarias/citología , Células Germinativas/citología , Factor Neurotrófico Derivado de la Línea Celular Glial/farmacología , Factor Inhibidor de Leucemia/farmacología , Testículo/citología , Animales , Bovinos , Recuento de Células , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Ensayo de Unidades Formadoras de Colonias , Cuerpos Embrioides/citología , Cuerpos Embrioides/efectos de los fármacos , Células Madre Embrionarias/efectos de los fármacos , Células Madre Embrionarias/metabolismo , Masculino , Células Madre Pluripotentes/citología , Células Madre Pluripotentes/efectos de los fármacos , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
To preliminarily clarify the mechanism of cardiac microvascular endothelial barrier function leading to heart failure, primary HMVEC-D cells were selected and cultured for amplification. The cells were infected with adenovirus vector containing the ADP-ribosylation factor 6 (Arf6) Q67L gene. Full-length and functional fragments of myeloid differentiation primary response 88 and ARF nucleotide-binding site opener genes were established and transfected into HEK293T cells. GTP-Arf6 pull-down experiment, fluorescent quantitative real-time PCR, immuno-coprecipitation, and transendothelial electrical resistance analysis were conducted. Interleukin-1ß (IL-1ß) induced increase in vascular permeability, whereas inhibitor SC514 blocked IL-1ß-induced transfer of nuclear factor-κB into the nucleus, from the cytoplasm. Increase in amount of activated Arf6 promoted reduction in transendothelial electrical resistance. In addition, SecinH3 significantly inhibited increase in vascular permeability, and the progression of heart failure was significantly relieved. Cardiac microvascular endothelial barrier function can lead to heart failure. However, IL-1ß induced increase in vascular permeability, which nullified the function of cardiac microvascular endothelial barrier. These findings are closely related to the activation of the Arf6-VE-cadherin signaling pathway.
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Células Endoteliales/metabolismo , Microvasos/citología , Miocardio/citología , Factor 6 de Ribosilación del ADP , Factores de Ribosilacion-ADP/metabolismo , Permeabilidad Capilar/efectos de los fármacos , Permeabilidad de la Membrana Celular/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Células HEK293 , Insuficiencia Cardíaca/metabolismo , Insuficiencia Cardíaca/fisiopatología , Humanos , Interleucina-1beta/farmacologíaRESUMEN
This study investigated the association of tumor necrosis factor-α (TNF-α)-308, -238, and -863 polymorphisms with osteoarticular tuberculosis (OA-TB) prognosis in a Hebei population. Genomic DNA was extracted from venous blood samples of 120 OA-TB patients and 100 healthy volunteers. TNF-α-308, -238, and -863 were analyzed by PCR-restriction fragment length polymorphism; genotype and allele frequencies were calculated. Serum TNF-α level was significantly higher in OA-TB patients (283.16 ± 51.68 ng/L) than in control (122.54 ± 54.65 ng/L; P < 0.05). Higher frequency of TNF-α-308 GG genotype in healthy volunteers (91.0%) than in OA-TB patients (79.2%) indicated that it was a protective factor against OA-TB (OR = 0.405, 95%CI = 0.147-0.657, P = 0.007). Higher frequencies of TNF-α-308 GA genotype and TNF-α-308 allele (A) in OA-TB patients (20.8 and 10.4%, respectively) than in healthy volunteers (8.0 and 5.0%, respectively) indicated an association with increased risk of OA-TB (OR = 3.112, 95%CI = 1.520-6.343, P = 0.003; OR = 3.109, 95%CI = 1.676-6.538, P = 0.006; respectively). Haplotype association analysis of TNF-α polymorphisms (-308/-238/-863) showed a higher frequency of TNF-α AGA in OA-TB patients (12.1%) than in healthy volunteers (3.5%), indicating that it was a risk factor for OA-TB (OR = 4.201, 95%CI = 1.80-9.91, P = 0.010). TNF-α-308 G/A and TNF-α AGA (-308/-238/-863) were associated with a predisposition to OA-TB, which could aid clinical detection, prevention, and prognosis of OA-TB.
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Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Tuberculosis Osteoarticular/genética , Factor de Necrosis Tumoral alfa/genética , Adulto , Alelos , Femenino , Genotipo , Haplotipos , Humanos , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , Pronóstico , Factores de Riesgo , Tuberculosis Osteoarticular/patologíaRESUMEN
There is high incidence of periodontal disease in high-altitude environments; hypoxia may influence the proliferation and clone-forming ability of periodontal ligament stem cells (PDLSCs). The MAPK signaling pathway is closely correlated with cell proliferation, differentiation, and apoptosis. Thus, we isolated and cultured PDLSCs under hypoxic conditions to clarify the impact of hypoxia on PDLSC proliferation and the underlying mechanism. PDLSCs were separated and purified by the limiting dilution method and identified by flow cytometry. PDLSCs were cultured under hypoxic or normoxic conditions to observe their cloning efficiency. PDLSC proliferation at different oxygen concentrations was evaluated by MTT assay. Expression of p38/MAPK and MAPK/ERK signaling pathway members was detected by western blotting. Inhibitors for p38/MAPK or ERK were applied to PDLSCs to observe their impacts on clone formation and proliferation. Isolated PDLSCs exhibited typical stem cell morphological characteristics, strong abilities of globular clone formation and proliferation, and upregulated expression of mesenchymal stem cell markers. Stem cell marker expression was not statistically different between PDLSCs cultured under hypoxia and normoxia (P > 0.05). The clone number in the hypoxia group was significantly higher than that in the control (P < 0.05). PDLSC proliferation under hypoxia was higher than that of the control (P < 0.001). p38 and ERK1/2 phosphorylation in hypoxic PDLSCs was markedly enhanced compared to that in the control (P < 0.05). Either P38/MAPK inhibitor or ERK inhibitor treatment reduced clone formation and proliferation. Therefore, hypoxia enhanced PDLSC clone formation and proliferation by activating the p38/MAPK and ERK/MAPK signaling pathways.
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Sistema de Señalización de MAP Quinasas , Ligamento Periodontal/citología , Células Madre/citología , Adolescente , Adulto , Apoptosis , Diferenciación Celular , Hipoxia de la Célula , Proliferación Celular , Células Cultivadas , Humanos , Persona de Mediana Edad , Adulto JovenRESUMEN
Biological changes in Snail-overexpressed SGC7901 cells were studied by establishing a pEGFP-C1-Snail carrier. The significance of Snail in epithelial-mesenchymal transition (EMT) as well as the invasion and metastatic capacity of gastric cancer cells was also discussed; moreover, we attempted to verify the probable cancer stem cell characteristics of Snail-overexpressed cells. A pEGFP-C1-Snail eukaryotic expression plasmid was constructed and pEGFP-C1(-) and pEGFP-C1-Snail plasmids were extracted and transfected into SGC7901 cells using Lipofectamine 2000. Stably expressed SGC7901-N [control group containing pEGFP-C1(-)] and SGC7901-S (test group containing pEGFP-C1-Snail) cells were screened using a G418 resistance medium. Snail, E-cadherin, b-catenin, vimentin, and fibronectin gene and protein expressions were detected by real-time quantitative PCR, western blot, and immunofluorescence. Cell invasion and metastasis were tested by scratch test, invasion assay, and an adhesion experiment. The positive rate of aldehyde dehydrogenase-1 (ALDH-1) expression was analyzed by flow cytometry. The results indicated the occurrence of EMT, accompanied by morphological changes in the cells and a weakening of the cell adhesion capacity. We also observed a decrease in the expression of epithelial markers E-cadherin and b-catenin and an increase in mesenchymal (Snail and vimentin) marker expression. Moreover, the cells showed increased invasiveness and metastatic capacity, and decreased proliferative ability. Moreover, the Snail-treated SGC7901 cells moved towards the scratch and produced fewer clones compared to the control cells. Owing to its capacity for self-renewal, SGC7901-S cells produced new clones and expressed ALDH-1. Therefore, we concluded that Snail overexpression induced EMT and endowed cells with tumor stem cell characteristics.
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Células Epiteliales/metabolismo , Transición Epitelial-Mesenquimal/genética , Regulación Neoplásica de la Expresión Génica , Células Madre Neoplásicas/metabolismo , Factores de Transcripción de la Familia Snail/genética , Familia de Aldehído Deshidrogenasa 1 , Antígenos CD , Cadherinas/genética , Cadherinas/metabolismo , Adhesión Celular , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Células Epiteliales/patología , Fibronectinas/genética , Fibronectinas/metabolismo , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patología , Genes Reporteros , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Isoenzimas/genética , Isoenzimas/metabolismo , Células Madre Neoplásicas/patología , Plásmidos/química , Plásmidos/metabolismo , Retinal-Deshidrogenasa/genética , Retinal-Deshidrogenasa/metabolismo , Transducción de Señal , Factores de Transcripción de la Familia Snail/metabolismo , Transfección , Transgenes , Vimentina/genética , Vimentina/metabolismo , beta Catenina/genética , beta Catenina/metabolismoRESUMEN
DIO3 gene encoding type 3 iodothyronine deiodinase is an imprinted gene, located in the DLK1-DIO3 (delta-like 1 homolog-type 3 iodothyronine deiodinase) imprinted domain, and is potentially involved in degrading excessive amounts of thyroid hormone to protect embryogenesis. However, the underlying regulatory mechanism of the imprinted DIO3 gene expression during fetal and neonatal development in goats has not been elucidated. In this study, we explored the DNA methylation patterns of the caprine DIO3 intragenic CpG island and quantified gene expression level in six tissues from Chinese Nanjiang Yellow 3-day old kids. The expression of the DIO3 gene was determined using quantitative reverse transcription-polymerase chain reactions (qRT-PCRs), while the identification of methylation patterns was determined using bisulfite-sequencing PCRs. Modest, and non-significant (P > 0.05), methylation patterns were noted for the DIO3 CpG island methylation in the brain, heart, liver, kidney, lung, and longissimus dorsi tissues (ranging from 26.48 to 34.92%). The expression level of the DIO3 mRNA was significantly higher (P < 0.05) in the liver tissue than in the other five tissues. Pearson's correlation analysis revealed that there was no significant relationship between methylation and gene expression (P > 0.05), which indicated that the expression of the caprine DIO3 gene was likely modified by other regulatory elements. This study identified DNA methylation and expression patterns of the DIO3 gene in goats and provided insights into further regulatory mechanisms of expression and imprinting in the DLK1-DIO3 domain.
Asunto(s)
Metilación de ADN , Epigénesis Genética , Regulación del Desarrollo de la Expresión Génica , Cabras/genética , Yoduro Peroxidasa/genética , Hígado/enzimología , Animales , Animales Recién Nacidos , Encéfalo/enzimología , Encéfalo/crecimiento & desarrollo , Islas de CpG , Cabras/crecimiento & desarrollo , Cabras/metabolismo , Corazón/crecimiento & desarrollo , Yoduro Peroxidasa/metabolismo , Riñón/enzimología , Riñón/crecimiento & desarrollo , Hígado/crecimiento & desarrollo , Pulmón/enzimología , Pulmón/crecimiento & desarrollo , Músculo Esquelético/enzimología , Músculo Esquelético/crecimiento & desarrolloRESUMEN
miRNA-203 is involved in the development and progression of various types of cancer. However, its role in cervical cancer remains unclear. The aim of this study was to investigate the effect of miRNA-203 on the proliferation and migration of HeLa cervical cancer cells, as well as survivin expression in these cells. A miRNA-203 primer probe was designed according to a sequence obtained from NCBI. The expression of miRNA-203 in cervical epithelial cells and cervical cancer cells was detected by quantitative reverse transcriptase-polymerase chain reaction. The miRNA-203 expression pattern was compared between these two cell lines. The cervical cancer cells were transfected with miRNA-203 mimic or inhibitor to determine their effects on proliferation and migration. The expression of the miRNA-203 target protein (survivin) was analyzed by western blot. Cervical cancer cells showed reduced miRNA-203 expression compared to cervical epithelial cells. Transfection of miRNA-203 mimic upregulated the expression of miRNA-203, suppressed cell proliferation and migration, and downregulated survivin expression (P < 0.05). However, downregulation of miRNA-203 expression did not affect proliferation, migration, and survivin expression in cervical cancer cells (P > 0.05). In conclusion, upregulation of miRNA-203 in cervical cancer cells inhibits the proliferative and migratory capacities of these cells by downregulating the expression of survivin.
Asunto(s)
MicroARNs/genética , Neoplasias del Cuello Uterino/genética , Movimiento Celular/genética , Proliferación Celular , Femenino , Regulación Neoplásica de la Expresión Génica , Células HeLa , Humanos , Proteínas Inhibidoras de la Apoptosis/metabolismo , MicroARNs/metabolismo , Survivin , Neoplasias del Cuello Uterino/patologíaRESUMEN
The growth trait is important in poultry production. We analyzed the association between single nucleotide polymorphisms (SNPs) in the Myf5 and MyoG gene and Bian chicken growth traits. SNPs in candidate genes of the Bian chickens were detected by the polymerase chain reaction-single strand conformation polymorphism method. Two mutation loci and six genotypes were identified in each candidate gene. In terms of growth traits, least square analysis showed that the FF genotype of the MyoG was the advantage genotype and the IJ genotype of the Myf5 was the disadvantage genotype for growth trait in Bian chicken. Correlation analysis suggested that the different combination genotypes between Myf5 and MyoG genes had a significant effect on growth traits in Bian chickens. The result suggested that MyoG and Myf5 genes can be used in marker-assisted selection for improving the growth trait in Bian chicken.