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BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is a kind of cancer with heterogeneous biological characteristics, which is affected by a complex network of gene interactions. Identification of molecular biomarkers paves the way for individualized therapy based on gene expression profiles, which can overcome the heterogeneity of ESCC. METHODS: In this study, GSE20347, GSE23400 and GSE45670 datasets were retrieved from Gene Expression Omnibus (GEO) database, and the overlapping differentially expressed genes (DEGs) in three datasets were screened. Then the overlapping DEGs function was annotated by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway-enrichment analysis. The prognostic value of the top five KEGG pathway-related genes were further validated in The Cancer Genome Atlas (TCGA) database. After extensive statistical analysis, four genes (CDC25B, CXCL8, FZD6 and MCM4) were identified as potential prognostic markers. Among the four candidate genes, the prognostic value of FZD6 in ESCC patients has not been evaluated. Therefore, we finally used immunohistochemistry method to evaluate the effect of FZD6 on the prognosis of patients with ESCC. Additionally, we detected the expression level of FZD6 in ESCC cell line and normal esophageal epithelial cell line, and observed the cell viability of ESCC cell line after FZD6 knockdown. RESULTS: The results showed that the overexpression of FZD6 predicted poor overall survival (OS) (P = 0.005) and progression-free survival (PFS) (P = 0.004) in ESCC patients. COX regression analysis showed that N stage (P = 0.026) and FZD6 expression level (P = 0.001) were independent prognostic factors of OS for ESCC patients. Furthermore, compared with normal esophageal epithelial cell line, the up-regulation of FZD6 was detected in ESCC cell line. Knockdown of FZD6 could significantly inhibit the proliferation of ESCC cells (P < 0.001). CONCLUSION: CDC25B, CXCL8, FZD6 and MCM4 were screened as candidate genes for prognosis assessment of patients with ESCC. The prognostic role of FZD6 in ESCC patients was confirmed in current study.
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Neoplasias Esofágicas/genética , Carcinoma de Células Escamosas de Esófago/genética , Receptores Frizzled/genética , Línea Celular Tumoral , Supervivencia Celular/genética , Femenino , Técnicas de Silenciamiento del Gen , Humanos , Inmunohistoquímica , Técnicas In Vitro , Interleucina-8/genética , Masculino , Persona de Mediana Edad , Componente 4 del Complejo de Mantenimiento de Minicromosoma/genética , Supervivencia sin Progresión , Modelos de Riesgos Proporcionales , ARN Interferente Pequeño , Tasa de Supervivencia , Regulación hacia Arriba , Fosfatasas cdc25/genéticaRESUMEN
OBJECTIVE: This study reviewed the distribution of each tumor stage and each type of initial treatment modality among patients with primary hepatocellular carcinoma (HCC) treated at a tertiary tumor hospital between January 2003 and October 2013. METHODS: Baseline data of patients with primary hepatocellular carcinoma treated between January 2003 and October 2013 were retrospectively collected. Tumor stage was determined according to the Barcelona Clinic Liver Cancer (BCLC) staging system and Hong Kong Clinic Liver Cancer (HKLC) staging system. RESULTS: A total of 6241 patients with primary hepatocellular carcinoma were included in the analysis. In accordance with the BCLC, 28.9% of patients were in stage 0/A, 16.2% in stage B, 53.6% in stage C, and 1.3% in stage D. According to the HKLC stage system, 8.4% patients were in stage I, 1.5% in stage IIa, 29.0% in stage IIb, 10.0% in stage IIIa, 33.6% in stage IIIb, 3.4% in stage IVa, 2.5% in stage IVb, 0.2% in stage Va, and 11.4% in stage Vb. Treatment modalities applied to this patient group were as follows: 33.3% of patients underwent hepatic resection, 36.7% underwent transarterial chemoembolization (TACE), 2.2% underwent radiotherapy, 0.9% underwent local ablated therapy, 8.8% underwent systemic chemotherapy, 4.2% underwent traditional herbal medicine therapy, 0.1% underwent targeted drug therapy, and 13.8% received no treatment. Hepatic resection was the most frequent therapy for patients with BCLC 0/A/B disease, and TACE was the initial therapy for patients with BCLC C disease. In the Hong Kong Clinic Liver Cancer staging system, the main treatments for HKLC I to IIIb disease is hepatic resection and TACE. Systemic chemotherapy was the initial therapy for patients with HKLC IVa/IVb disease. Most HKLC Va/Vb patients received traditional Chinese medicine treatment. CONCLUSION: Prevalence of stage BCLC B and C disease was high among our hepatocellular carcinoma patients. In Hong Kong Clinic Liver Cancer staging system, HKLC I to IIIb disease was high among our HCC patients. Hepatic resection and TACE are initial therapies.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Carcinoma Hepatocelular/patología , Quimioembolización Terapéutica , Hepatectomía , Neoplasias Hepáticas/patología , Carcinoma Hepatocelular/terapia , Terapia Combinada , Femenino , Estudios de Seguimiento , Humanos , Neoplasias Hepáticas/terapia , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Estudios Retrospectivos , Tasa de SupervivenciaRESUMEN
The aim of this study was to examine the subtype distribution of human papilloma virus (HPV) in women in the Shaanxi Province of China. A DNA chip, along with polymerase chain reaction amplification and reverse dot blot technology, was adopted to analyze the HPV genotypes of 22,937 cases of cervical cell specimens. The HPV infection rate was 18.70%, wherein high-risk, low-risk, and high- and low-risk multiple infection rates were 15.75, 2.96 and 1.91%, respectively. High-risk infections accounted for 84.20% of total infections. The rate of HPV infection in women with rural residence, high school education or less, a low income, or age over 40 years was significantly higher than that in the control group (negative HPV infection women). Of the 18 detected high-risk HPV subtypes, the most common in single infections were, in the order of prevalence, HPV16, 58, 18, 52, 33, and 56. For multiple high-risk infections, the most common subtypes in the order of prevalence were HPV16, 52, 58, 18, 56, and 33. Age was a factor in the rate of infection, as the 41-50-year age group had a significantly higher risk of infection than the other groups (P < 0.05). In multiple infections, double infections were common, accounting for 77.10% of multiple infections, and triple or more infections were more common in women aged 51-60 years. In Shaanxi Province, high-risk HPV infection in women was mainly attributed to rural residence, age over 40 years, low income, and low education level.
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Técnicas de Genotipaje/métodos , Papillomaviridae/genética , Adulto , Distribución por Edad , China , Femenino , Genotipo , Humanos , Persona de Mediana Edad , Infecciones por Papillomavirus/virología , Factores de Riesgo , Adulto JovenRESUMEN
MicroRNA (miRNA) deregulation has been previously linked to the initiation and development of breast cancer. Although miR-99a is aberrantly expressed in many types of cancers, including breast cancer, the serum miR-99a expression level in breast cancer and its clinical significance remains unknown. Blood samples were obtained from 72 patients with breast cancer and 40 healthy volunteers, and subjected to real-time polymerase chain reaction to evaluate the level of expression of serum miR-99a in the study participants. Furthermore, we investigated the association between serum miR-99a and the clinical outcome of breast cancer. Serum miR-99a expression was significantly downregulated in patients with breast cancer, compared to that in healthy controls (P < 0.01). Moreover, the serum miR-99a was correlated with various clinical parameters of breast cancer, including lymph node metastasis (P = 0.0194), distant metastasis (P = 0.0037), Ki67 intensity (P = 0.0164), TNM stage (P = 0.0096), and histological grade (P = 0.0051) of cancer. Additionally, breast cancer patients displaying lower miR-99a levels showed poorer overall survival rates (P = 0.0411). The serum miR-99a level was also found to be an independent risk factor for breast cancer (hazard ratio = 3.176, 95% confidence interval = 1.543-7.360, P = 0.023). Our data indicated that serum miR-99a expression was downregulated in breast cancer patients; moreover, this downregulation was associated with poor prognosis, suggesting that serum miR-99a could function as a tumor suppressor in breast cancer.
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Neoplasias de la Mama/diagnóstico , Neoplasias de la Mama/genética , Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , Adulto , Neoplasias de la Mama/sangre , Neoplasias de la Mama/mortalidad , Estudios de Casos y Controles , Femenino , Humanos , Antígeno Ki-67/sangre , Antígeno Ki-67/genética , Metástasis Linfática , MicroARNs/sangre , Persona de Mediana Edad , Clasificación del Tumor , Estadificación de Neoplasias , Pronóstico , Receptor ErbB-2/genética , Receptor ErbB-2/metabolismo , Receptores de Estrógenos/genética , Receptores de Estrógenos/metabolismo , Receptores de Progesterona/genética , Receptores de Progesterona/metabolismo , Factores de Riesgo , Análisis de SupervivenciaRESUMEN
The aim of this study was to explore the molecular mechanism by which all-trans retinoic acid (ATRA) prevents type 1 diabetes mellitus (T1DM). Fifty ICR mice were randomly assigned to three groups: prevention group [N = 20; mice received 10 mg/kg ATRA daily for 5 days and then 60 mg/kg streptozotocin (STZ) for 5 days]; diabetic group (N = 20, mice received 95% sterile peanut oil and 5% dimethyl sulfoxide for 5 days and then 60 mg/kg STZ for 5 days); and control group (N = 10, mice received 95% sterile peanut oil and 5% dimethyl sulfoxide for 5 days and then citrate buffer for 5 days). Blood glucose was measured using blood glucose test strips and serum insulin was measured by radioimmunoassay. Islets cell morphology was assessed by microscopy and ELISA was used to measure the serum levels of interferon gamma (IFN-γ) and interleukin 4 (IL- 4). In the prevention group, blood sugar levels were found to be reduced and serum insulin levels increased compared with the levels in the diabetic group (P < 0.05), indicating that ATRA prevented the STZ-induced damage to islet cells. Meanwhile, ATRA was shown to decrease the levels of IFN-γ and increase the levels of IL-4 as well as the IFN-γ/IL-4 ratio in STZ-treated animals (P < 0.05). These findings suggest that ATRA prevents the recurrence of autoimmune insulitis. This study demonstrated that ATRA effectively prevents the progression of T1DM in a murine model of the disease by reducing IFN-γ levels and increasing IL-4 levels.
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Diabetes Mellitus Experimental/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Interferón gamma/sangre , Interleucina-4/sangre , Tretinoina/uso terapéutico , Animales , Diabetes Mellitus Experimental/prevención & control , Hipoglucemiantes/administración & dosificación , Hipoglucemiantes/farmacología , Islotes Pancreáticos/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos ICR , Tretinoina/administración & dosificación , Tretinoina/farmacologíaRESUMEN
This study aimed to investigate human papilloma virus (HPV) genotypes among women with cervical lesions in Shaanxi Province, China, to obtain information regarding cervical lesion prevention and treatment. The study included 4508 HPV-positive subjects; cervical swab specimens were collected and tested for HPV infection status and HPV genotypes using polymerase chain reaction and reverse dot-blot hybridization. Women positive for HPV with cervical lesions, including chronic cervicitis, cervical intraepithelial neoplasia, and cervical squamous cell carcinoma (SCC), were examined; HPV-positive women with no cervical lesions were controls. Data were pooled and weighted estimates have been presented. For women with no cervical lesions and positive for one HPV genotype, HPV 52, 16, 58, 81, 33, and 56 were the most common; for multiple-HPV genotype infection, HPV 16, 52, 6, 18, 58, and 66 were the most common. Collectively, HPV 16, 58, 52, 18, 33, and 81 were the most common in women with cervical lesions. HPV 16 comprised 26.71% of single-genotype and 15.64% of multiple-genotype infections. The proportion of HPV-16-positive cases was 29.15%, which was the highest among all HPV genotypes (P < 0.01). Single-HPV genotype infection was the most common in cervical HPV infection (77.48%); infection with two HPV genotypes comprised 72.22% of multiple-genotype infections. The proportion of single-low-risk HPV genotype infections decreased with increase in cervical lesion severity; there were no single- or multiple-low-risk genotype HPV infections in cervical SCC patients. The proportion of multiple-genotype HPV infections with at least one high-risk genotype increased with cervical lesion severity.
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Papillomaviridae/clasificación , Papillomaviridae/genética , Neoplasias del Cuello Uterino/virología , Adulto , Anciano , Carcinoma de Células Escamosas/virología , China , ADN Viral/genética , Femenino , Genotipo , Papillomavirus Humano 16/clasificación , Papillomavirus Humano 16/genética , Humanos , Persona de Mediana Edad , Infecciones por Papillomavirus/complicaciones , Infecciones por Papillomavirus/virología , Prevalencia , Adulto JovenRESUMEN
Lipasin has recently been demonstrated to be involved in lipid metabolism. In this study, two specific primers were used to amplify the lipasin open reading frame from porcine liver tissue. The polymerase chain reaction product was cloned to a pGEM®-T Easy Vector, digested by SalI and NotI, and sequenced. The lipasin fragment was then cloned to a pET21(b) vector and digested by the same restriction enzyme. The recombinant plasmid was transferred to Escherichia coli (BL21), and the lipasin protein was induced with isopropyl-ß-D-thiogalactopyranoside. The protein obtained was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blotting. A pET-lipasin prokaryotic recombinant expression vector was successfully constructed, and a 25.2-kDa protein was obtained. This study provides a basis for further research on the biological function of porcine lipasin.
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Metabolismo de los Lípidos/genética , Proteínas Recombinantes de Fusión/biosíntesis , Porcinos/genética , Proteína 8 Similar a la Angiopoyetina , Proteínas Similares a la Angiopoyetina , Animales , Clonación Molecular , Escherichia coli/genética , Regulación de la Expresión Génica , Hígado/metabolismo , Hormonas Peptídicas/genética , Proteínas Recombinantes de Fusión/genéticaRESUMEN
To understand the effects of disease-resistant maize varieties and new cropping systems on the population of Curvularia lunata, 52 isolates of C. lunata were collected in China from 2011 to 2013. The isolates were analyzed in terms of phylogenetic relationships, morphology, and pathogenicity. Phylogenetic analysis showed that the 52 isolates clustered into 2 distinct clusters with further subdivisions, suggesting the emergence of new genetic divergence within C. lunata. Results of morphology and pathogenicity analyses demonstrated that there were significant differences among these isolates: 27 isolates were classified as fast growing, 5 as slow growing, and 20 as moderate growing. Three isolates had white-colored colonies, 13 had yellowish green-colored colonies, and the remaining isolates had dark green-colored colonies. Furthermore, conidiation rates were assessed: 30 isolates were characterized as having low conidiation rates, 15 as having medium conidiation rates, and the remaining 7 isolates as having high conidiation rates. Eleven of the isolates appeared to be strongly pathogenic against maize, 15 isolates proved to be weakly pathogenic against maize, and the remaining isolates were regarded to be moderately pathogenic. Interestingly, correlation analysis demonstrated a negative correlation between the growth rate and the pathogenicity of the isolates, while a positive correlation was observed between the conidiation rate and the pathogenicity. No correlation was observed between the colony color and the pathogenicity of the isolates.
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Ascomicetos/genética , Ascomicetos/patogenicidad , Enfermedades de las Plantas/microbiología , Zea mays/microbiología , China , Filogenia , VirulenciaRESUMEN
We explored the molecular mechanism of the regulation of vacuolar-type-H+-ATPase B1 (VHAB1) in elvers in the response to salinity. The full-length cDNA of VHAB1 in Anguilla marmorata (designated as AmVHAB1), which was 1741 base pairs (bp) in length, was found to encompass a 1512-bp open reading frame encoding a polypeptide with 503 amino acids (55.9 kDa), an 83-bp 5'-untranslated region, and a 146-bp 3'-untranslated region. The mRNA and protein expression levels of AmVHAB1 in the gill were evaluated at different time points (0, 1, 3, 6, 12, 24, 48, 72, and 96 h, and 15 days) during the exposure to various salinity levels (0, 10, and 25). The results indicated that the expression levels of AmVHAB1 mRNA in the gill significantly increased and reached the highest level at 1 h exposure in the brackish water (BW, 10) group and at 6 h exposure in the seawater (SW, 25) group. The salinity level affected the relative expression level of AmVHAB1 mRNA in the gill, which was increased by approximately 44-fold in the SW group when compared with that in fresh water. Immunoblotting analysis showed that VHA expression was significantly higher in the BW and SW groups, with the highest expression level was detected at 96 h exposure. We found that the AmVHAB1 gene in elvers from A. marmorata plays an important role in the adaptation to seawater.
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Anguilla/genética , Branquias/enzimología , ATPasas de Translocación de Protón Vacuolares/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Regulación Enzimológica de la Expresión Génica , Immunoblotting , Datos de Secuencia Molecular , Filogenia , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Aguas Salinas , Agua de Mar , Factores de Tiempo , ATPasas de Translocación de Protón Vacuolares/química , ATPasas de Translocación de Protón Vacuolares/metabolismoRESUMEN
Rspo1 belongs to the Rspo family, which is composed of 4 members (Rspo1-4) that share 40 to 60% sequence homology and similar domain organizations, and regulate the WNT signaling pathway via a common mechanism. Rspo1 plays a key role in vertebrate development and is an effective mitogenic factor of gastrointestinal epithelial cells. We report the cloning of chicken Rspo1 and its gene expression distribution among tissues. It contained an open reading frame of 783 bp encoding a protein of 260 amino acids, and its molecular weight was predicted to be 28.80 kDa. Reverse transcription-polymerase chain reaction-based gene expression analysis indicated that chicken Rspo1 was highly expressed in the stomach muscle tissue, but was expressed at low levels in the lung, brain, jejunum, cecum, ileum, spleen, pancreas, kidney, and glandular stomach. These results suggest that Rspo1 plays a major role in muscular immune protection.
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Pollos/genética , Trombospondinas/genética , Animales , Clonación Molecular/métodos , Femenino , Masculino , Análisis de Secuencia de ADN , Distribución Tisular , Vía de Señalización WntRESUMEN
Gene mutation plays an important role in molecular biological studies. A highly efficient one-step polymerase chain reaction-based mutagenesis technique for site-directed mutagenesis was developed in this study. One complementary pair of primers was designed that contained the desired mutations in the middle of the primers. The amplification products of mutation were amplified using a high-fidelity DNA polymerase and the original plasmid templates were digested by DpnI. This method was successfully used to introduce mutations in two different-sized plasmids (12 and 6 kb) with high efficiency. The results indicate that this technique can be widely used to introduce any plasmid mutations quickly and efficiently.
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ADN Polimerasa Dirigida por ADN/química , Mutagénesis Sitio-Dirigida/métodos , Plásmidos/genética , Secuencia de Bases , Análisis Mutacional de ADN , Proteínas Fluorescentes Verdes/genética , Reacción en Cadena de la PolimerasaRESUMEN
The influence of ruminal acidosis on ruminal microbiology and metabolite production has received considerable attention, but little is known regarding the systemic manifestations that arise from ruminal acidosis. Lipopolysaccharide (LPS) is released in the gastrointestinal tract upon ingestion of high-grain or high-fat diets, and it has been implicated in the etiology of multiple energy- and lipid-related metabolic disturbances in ruminants. The liver plays a crucial role in the acute phase response to intruding pathogens. The effect of blood LPS in subacute ruminal acidosis on lipid metabolism in the liver has not been established. In this study, cell cultures were photographed using an inverted microscope. We observed that hepatocytes changed their morphologies from irregular triangle to circular (contraction) shapes; the number of contracted cells increased with the increasing LPS doses. This suggests that LPS can promote cell contraction and take off the wall, ultimately leading to cell apoptosis. With changes in LPS exposure, hepatocyte number also changes. We explored lipid metabolism in the liver using quantitative reverse transcription-polymerase chain reaction to detect the expression of key lipid metabolism enzymes in hepatocytes. We found that Toll-like receptor 4 signaling pathway mediated by LPS could attenuate mRNA expression of fatty acid synthesis genes and increase the expression of fatty acid transport genes in primary hepatocytes following LPS treatment in dairy cows.
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Expresión Génica/inmunología , Hepatocitos/metabolismo , Metabolismo de los Lípidos , Lipopolisacáridos/farmacología , Animales , Bovinos , Forma de la Célula/inmunología , Células Cultivadas , Femenino , Regulación de la Expresión Génica/inmunología , Hepatocitos/inmunología , Cultivo Primario de Células , ARN Mensajero/genética , ARN Mensajero/metabolismo , Transducción de Señal , Receptor Toll-Like 4/metabolismoRESUMEN
Transcriptome sequencing technology has been applied in the development and discovery of single nucleotide polymorphism (SNP) markers in fish. In this study, a panel of 120 expressed sequence tag (EST)-derived SNPs was selected by several selection filters from the resultant EST library of Odontobutis potamophila using Illumina Sequencing. In total, 37 SNPs from 120 putative SNPs were considered as the true SNPs using Sanger sequencing. For each SNP locus of 30 individuals of one wild population of O. potamophila that was successfully calculated, the number of alleles per locus was 2 with an observed heterozygosity of 0.0000-0.9000 and an expected heterozygosity of 0.1000-0.5263. A total of 33 loci conformed to Hardy-Weinberg equilibrium (HWE), and 4 loci deviated from HWE after Bonferroni correction. These 33 SNP markers will benefit the studies of population genetic structure, population evolution analysis, and construction of a high-density linkage map of O. potamophila.
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Perciformes/genética , Animales , Mapeo Cromosómico , Etiquetas de Secuencia Expresada , Polimorfismo de Nucleótido Simple , Análisis de Secuencia de ADN , TranscriptomaRESUMEN
Studying natural variation in rice resistance genes of cultivated and wild rice relatives can predict resistance stability to rice blast fungus. In the present study, the protein coding regions of the rice R gene Pi-d2 in 35 rice accessions, including Oryza sativa L. subsp. indica Kato (Aus), indica (IND), temperate japonica (TEJ), tropical japonica (TRJ), aromatic (ARO); subgroups of Oryza sativa; 6 accessions of wild rice varieties; O. nivara; and O. rufipogon were analyzed. A total of 13 nucleotide differences were found in the open reading frames (ORFs) of Pi-d2. Translation of these ORFs revealed 9 variants; 3 were novel Pi-d2 variants. Variants H2 and H5 were identified in accessions of cultivated rice and O. nivara, H1, H3, H4, H6, and H8 were only identified in cultivated rice. H2 and H5 were the common types of IND and O. nivara, H8 was the common type of TRJ and AUS, H6 was the specific type of AUS, and H3 was the specific type of ARO. H7 and H9 were specific haplotypes of O. nivara and O. rufipogon, respectively. These findings demonstrate that Pi-d2 variants are useful indicators for each subgroup, and Pi-d2 is an ancient gene that predates speciation of rice subgroups.
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Resistencia a la Enfermedad/genética , Genes de Plantas , Variación Genética , Proteínas de la Membrana/genética , Oryza/genética , Enfermedades de las Plantas/genética , Proteínas de Plantas/genética , Alelos , Aminoácidos/química , Productos Agrícolas/genética , ADN de Plantas/análisis , Evolución Molecular , Marcadores Genéticos , Genoma de Planta , Haplotipos , Magnaporthe , Proteínas de la Membrana/fisiología , Sistemas de Lectura Abierta , Filogenia , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/fisiología , Polimorfismo de Nucleótido Simple , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/fisiología , Especificidad de la EspecieRESUMEN
The association between the microsomal epoxide hydrolase 1 gene (EPHX1) Tyr113His polymorphism and lung cancer and breast cancer risk has been reported in many recent studies, but there is no consensus among the results. Thus, we examined the association between the EPHX1 Tyr113His polymorphism and lung cancer through a meta-analysis. A comprehensive literature search was performed using the Pubmed and Embase databases. Odds ratios with 95% confidence intervals were used to assess the strength of associations. Our meta-analysis suggested that the Tyr113His polymorphism was associated with lung cancer risk in Asians under 3 genetic models, including a C vs T, CC vs TT, and recessive model. However, the risk was decreased in Caucasians under the genetic models, including a C vs T, CC vs TT, or CT vs TT, dominant, and recessive model. In contrast, there was no association with breast cancer risk for any of the genetic models. Our meta-analysis suggested that the EPHX1 Tyr113His polymorphism may be a risk factor for lung cancer in Asians, whereas it may be a decreased risk factor among Caucasians. However, this polymorphism was not found to be associated with breast cancer.
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Neoplasias de la Mama/genética , Epóxido Hidrolasas/genética , Estudios de Asociación Genética , Neoplasias Pulmonares/genética , Polimorfismo de Nucleótido Simple , Alelos , Sustitución de Aminoácidos , Estudios de Casos y Controles , Femenino , Genotipo , Humanos , Masculino , Oportunidad Relativa , Sesgo de PublicaciónRESUMEN
Lactoferrin (Lf) is an iron-binding glycoprotein that is produced by mucosal epithelial cells in mammals. Lf has non-immune natural defense functions and biological functions in addition to and distinct from its role in regulating inflammatory responses. Lf also improved some physiological and immunological parameters. Lf is a biomarker for monitoring medical treatment in inflammatory bowel diseases. Current LF research focuses on iron absorption, antimicrobial activity, and the modulation of iron metabolism during inflammation. No systematic research about Lf expression levels in mouse mammary glands during pregnancy and lactation exists. We investigated Lf mRNA expression levels in mouse mammary glands by collecting samples on days 1, 6, 12, and 18 of pregnancy and lactation (six mice per group). The expression levels of Lf mRNA were measured by semi-quantitative reverse transcription polymerase chain reaction using GAPDH as an internal control. Lf mRNA was not expressed in mammary glands on days 1, 6, and 12 of pregnancy, but it was expressed on day 18 (IOD: integrated optical density; Lf(IOD)/GAPDH(IOD) = 0.46). Lf expression levels were higher during lactation stages than during pregnancy stages, and it stabilized at 0.71-0.73 (Lf(IOD)/GAPDH(IOD)) from day 1 to 12 of lactation; however, the difference was not significant (P > 0.05). At day 18 of lactation, Lf expression began to decline (Lf(IOD)/GAPDH(IOD) = 0.61), but the difference was not significant (P > 0.05). Based on these results, the variation in Lf expression levels during developmental stages may be related to its regulatory role in mouse mammary gland immunity.
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Células Epiteliales/metabolismo , Regulación del Desarrollo de la Expresión Génica , Lactancia/genética , Lactoferrina/genética , Glándulas Mamarias Animales/metabolismo , ARN Mensajero/genética , Animales , Células Epiteliales/citología , Células Epiteliales/inmunología , Femenino , Genes Esenciales , Gliceraldehído 3-Fosfato Deshidrogenasa (NADP+)/genética , Gliceraldehído 3-Fosfato Deshidrogenasa (NADP+)/metabolismo , Hierro/metabolismo , Lactancia/inmunología , Lactancia/metabolismo , Lactoferrina/inmunología , Lactoferrina/metabolismo , Glándulas Mamarias Animales/citología , Glándulas Mamarias Animales/inmunología , Ratones , Embarazo , ARN Mensajero/inmunología , ARN Mensajero/metabolismo , Factores de TiempoRESUMEN
Microsomal epoxide hydrolase 1 (EPHX1) is an important biological phase II metabolic enzyme that is extensively involved in the metabolism of diverse environmental carcinogens such as polycyclic aromatic hydrocarbons and heterocyclic amines. Many articles have reported the association between EPHX1 (Tyr113His and His139Arg) polymorphisms and esophageal cancer risk, but the results are controversial. This study aimed to identify the association between EPHX1 (Tyr113His and His139Arg) polymorphisms and esophageal cancer risk by meta-analysis. The odds ratio (OR) with 95% confidence interval (95%CI) was used to evaluate the strength of the associations. Heterogeneity was estimated by the chi-square-based Q-statistic test and the P value. Meanwhile, the random-effect or fixed-effect model was used according to the between-study heterogeneity. Begg's funnel plot and the Egger test were performed to assess the publication bias of articles. Finally, 8 case-control studies involving 1158 cases and 1868 controls for the Tyr113His polymorphism and 7 case-control studies involving 901 cases and 1615 controls for the His139Arg polymorphism were included in this meta-analysis. Meta-analysis showed that the Tyr113His polymorphism was a stronger power trend towards risk for esophageal cancer using a recessive model (CC versus CT+TT, OR = 1.204, 95%CI = 1.001-1.450, P = 0.049). However, no significant associated risk was found between the His139Arg polymorphism and esophageal cancer. These findings suggest that the Tyr113His polymorphism might be a stronger power trend towards risk for esophageal cancer. However, no evidence was found for the association between the EPHX1 His139Arg polymorphism and esophageal cancer risk.
Asunto(s)
Epóxido Hidrolasas/genética , Neoplasias Esofágicas/genética , Predisposición Genética a la Enfermedad , Femenino , Estudios de Asociación Genética , Humanos , Masculino , Polimorfismo de Nucleótido Simple , Factores de RiesgoRESUMEN
This study aimed to compare the effects of lipopolysaccharide (LPS) on stearoyl-coenzyme A desaturase (SCD) gene expression in mouse primary hepatic cells. To obtain sufficient total RNA, primary hepatic cells were plated on 6-cm diameter-type collagen 1-coated dishes (1 x 106 cells per dish). The test was divided into 6 groups with 6 replications per group. The 6 groups were treated with the following volumes of LPS (0.1 mg/mL): 0, 1, 1.5, 2, 4, and 8 µL. The cells were cultured for 24 h, and the total RNA was extracted from samples. Reverse transcription polymerase chain reaction was used to analyze SCD mRNA levels. With increasing LPS amounts, the SCD mRNA expression first decreased and then increased slightly; the expression was the lowest in the 2-µL LPS condition. The SCD mRNA levels from the 4- and 8-µL LPS conditions were slightly higher than that from the 2-µL LPS condition, but the difference was not significant (P > 0.05). The SCD mRNA level from the 2-µL LPS condition was obviously lower than that from the 0-, 1-, and 1.5-µL LPS condition, and the differences were significant (P < 0.05), and the SCD mRNA levels from the 0-, 1-, and 1.5-µL LPS conditions were not significantly different (P > 0.05). The SCD mRNA levels from the 4- and 8-µL LPS conditions were obviously lower than those from the 0- and 1-µL LPS conditions, and the differences were significant (P < 0.05).
Asunto(s)
Hepatocitos/efectos de los fármacos , ARN Mensajero/biosíntesis , Estearoil-CoA Desaturasa/biosíntesis , Animales , Bovinos , Regulación de la Expresión Génica/efectos de los fármacos , Lipopolisacáridos/administración & dosificación , Ratones , Estearoil-CoA Desaturasa/genéticaRESUMEN
We amplified S14R protein gene cDNA of porcine, cloned it into a prokaryotic expression plasmid, and expressed it in Escherichia coli. A pair of primers was designed based on the cDNA sequence of the porcine S14R gene in GenBank. The target gene fragment from porcine liver tissue was amplified by RT-PCR. Confirmed by auto-sequencing, the target gene fragment was subcloned into an expression vector of pET28a. The pET28a-S14R construct was subsequently transformed into E. coli BL21 (DE3). This construct was verified by restriction endonuclease digestion and sequencing. Using isopropyl ß-D-1-thiogalactopyranoside induction, a new recombinant protein with the expected relative molecular mass of 24 kDa appeared. The result was identified by SDS-PAGE electrophoresis. Porcine S14R includes 549bp (GenBank No. JN793537), with an open reading frame of 549 bp coding 182 amino acids.
Asunto(s)
Receptores Citoplasmáticos y Nucleares/genética , Sus scrofa/genética , Secuencia de Aminoácidos , Animales , Clonación Molecular , Escherichia coli , Expresión Génica , Datos de Secuencia Molecular , Receptores Citoplasmáticos y Nucleares/biosíntesis , Receptores Citoplasmáticos y Nucleares/química , Análisis de Secuencia de ADN , Homología de Secuencia de AminoácidoRESUMEN
Interkeukin-8 (IL-8) is an important inflammatory mediator. It is an angiogenic factor associated with inflammation and carcinogenesis. To date, research on IL-8 has been limited to its role as an indicator of inflammation. There has been no systematic research concerning IL-8 expression levels in the mouse mammary gland during pregnancy and lactation. Mouse mammary gland samples were collected on days 1, 6, 12, 18 of pregnancy and of lactation (6 mice per group). The expression levels of IL-8 mRNA were measured by semi-quantitative RT-PCR, with GAPDH as an internal control. IL-8 mRNA was highly expressed on day 1 of pregnancy in the mouse mammary glands (IL-8(IOD)/GAPDH(IOD) = 1.68), and then suddenly declined; it reached 0.74 and 0.71 on days 6 and 12 of pregnancy. On day 18 of pregnancy, it started to increase (IL-8(IOD)/GAPDH(IOD) = 1.02). However, the expression levels of IL-8 mRNA were not significant during pregnancy. During lactation, IL-8 expression level was lower than during pregnancy, but it stabilized at 0.32-0.41 (IL-8(IOD)/GAPDH(IOD)) from day 1 to day 18 of lactation, although the difference was not significant. We suggest that the changes in IL-8 expression level during development is related to its regulatory role in mouse mammary gland immunity.