RESUMO
BACKGROUND: Poly-(ADP-Ribose)-Polymerase inhibitors (PARPi) were reported as radiosensitizers in non-small cell lung cancer (NSCLC) with wide-type epidermal growth factor receptor (EGFR), but the effects of radiation combined with PARPi were not investigated in EGFR-mutated NSCLC. Moreover, the underlying mechanisms were not well examined. This study aimed to study the efficacy of radiation combined with niraparib in EGFR-mutated NSCLC and explore their influence on the immune system. METHODS: Clone formation and apoptosis assay were conducted to explore the effects of niraparib and radiation. Immunofluorescence was conducted to detect the double-strand DNA breaks. Real-time PCR and immunoblotting were employed to evaluate the activation of STING/TBK1/TRF3 pathway and the expression levels of interferon ß, CCL5 and CXCL10. Immunocompetent mice model bearing with subcutaneous Lewis lung cancer was established to confirm the results in vivo. RESULTS: Niraparib and radiation were synergistic to inhibit tumor both in vitro and in vivo. Radiation plus niraparib could activate anti-tumor immunity, which appeared as increased CD8+ T lymphocytes and activated STING/TBK1/IRF3 pathway. CONCLUSION: PARPi not only as a radiosensitizer inhibited EGFR-mutated NSCLC tumor growth, but also cooperated with radiation to promote anti-tumor immune responses.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/terapia , Quimiorradioterapia/métodos , Genes erbB-1 , Indazóis/farmacologia , Neoplasias Pulmonares/terapia , Mutação , Piperidinas/farmacologia , Inibidores de Poli(ADP-Ribose) Polimerases/farmacologia , Radiossensibilizantes/farmacologia , Animais , Linfócitos T CD8-Positivos , Carcinoma Pulmonar de Lewis/imunologia , Carcinoma Pulmonar de Lewis/terapia , Carcinoma Pulmonar de Células não Pequenas/imunologia , Carcinoma Pulmonar de Células não Pequenas/mortalidade , Linhagem Celular Tumoral , Quimiocina CCL5/metabolismo , Quimiocina CXCL10/metabolismo , Quebras de DNA de Cadeia Dupla , Feminino , Imunofluorescência , Humanos , Sistema Imunitário/efeitos dos fármacos , Sistema Imunitário/efeitos da radiação , Imunocompetência , Fator Regulador 3 de Interferon/metabolismo , Interferon beta/metabolismo , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/mortalidade , Linfócitos do Interstício Tumoral , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Nucleares/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Tolerância a Radiação/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Proteínas Semelhantes à Proteína de Ligação a TATA-Box/metabolismo , Ensaio Tumoral de Célula-TroncoRESUMO
OBJECTIVE: Liver metastasis is one of the major causes of cancer-related death in patients with colorectal cancer (CRC). The purpose of this study was to identify specific molecules which are involved in colorectal liver metastasis (CRLM). MATERIALS AND METHODS: In this study, we employed TMT (tandem mass tags)-labeling combined with liquid chromatography-mass spectrometry technology to do comparative analyses of proteomics between the primary tumor specimens derived from colorectal cancer patients with or without liver metastasis. Pathway enrichment analyses were performed using DAVID database. The crucial molecules were identified through protein-protein interaction network. Immunohistochemistry (IHC) was employed to analyze the expression of THBS1 (thrombospondin-1) in CRC tissues. Finally, transwell cell migration and invasion assays were performed to explore the roles of THBS1 in CRC cell migration and invasion. RESULTS: We found that the expression of 311 proteins was dysregulated in CRLM using quantitative proteomics. Among these proteins, we identified FN1, TIMP1, THBS1, POSTN and VCAN as five crucial proteins in CRLM by analysis in silico. IHC assay revealed that increased THBS1 expression was significantly correlated with liver metastasis as well as poor prognosis of CRC patients. GEO data analysis also suggests that upregulated mRNA level of THBS1 is also associated with shorter overall survival of CRC patients. Moreover, THBS1 depletion inhibited migration and invasion of CRC cells through attenuating epithelial-mesenchymal transition. Co-expression analyses with TCGA data indicated that THBS1 is co-expressed with mesenchymal markers, including Vimentin, N-cadherin, Snail1 and Twist1 in CRC tissues. CONCLUSIONS: By collecting the omics data with functional studies, the present results reveal that THBS1 facilitates colorectal liver metastasis through promoting epithelial-mesenchymal transition. This understanding of molecular roles of THBS1 in CRLM may be promising to develop targeted therapies to prolong survival in CRC patients.
Assuntos
Neoplasias Colorretais/patologia , Transição Epitelial-Mesenquimal , Neoplasias Hepáticas/secundário , Trombospondina 1/fisiologia , Linhagem Celular Tumoral , Movimento Celular , Neoplasias Colorretais/mortalidade , Humanos , Imuno-Histoquímica , Invasividade Neoplásica , Mapas de Interação de ProteínasRESUMO
Nitric oxide synthase (NOS) produces nitric oxide (NO) by catalyzing the conversion of l-arginine to l-citrulline, with the concomitant oxidation of nicotinamide adenine dinucleotide phosphate. Recently, various studies have verified the importance of NOS invertebrates and invertebrates. However, the NOS gene family in the oriental river prawn Macrobrachium nipponense is poorly understood. In this study, we cloned the full-length NOS complementary DNA from M. nipponense (MnNOS) and characterized its expression pattern in different tissues and at different developmental stages. Real-time quantitative polymerase chain reaction (RT-qPCR) showed the MnNOS gene to be expressed in all investigated tissues, with the highest levels observed in the androgenic gland (P < 0.05). Our results revealed that the MnNOS gene may play a key role in M. nipponense male sexual differentiation. Moreover, RT-qPCR revealed that MnNOS mRNA expression was significantly increased in post-larvae 10 days after metamorphosis (P < 0.05). The expression of this gene in various tissues indicates that it may perform versatile biological functions in M. nipponense.
Assuntos
Proteínas de Artrópodes/genética , Regulação da Expressão Gênica no Desenvolvimento , Óxido Nítrico Sintase/genética , Palaemonidae/genética , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/metabolismo , Sequência de Bases , China , Clonagem Molecular , Conservação dos Recursos Naturais , DNA Complementar/genética , DNA Complementar/metabolismo , Embrião não Mamífero , Feminino , Larva/genética , Larva/crescimento & desenvolvimento , Masculino , Óxido Nítrico Sintase/metabolismo , Técnicas de Amplificação de Ácido Nucleico , Especificidade de Órgãos , Palaemonidae/classificação , Palaemonidae/crescimento & desenvolvimento , Filogenia , Rios , Alinhamento de Sequência , Homologia de Sequência de AminoácidosRESUMO
Feminization-1 homolog b (Fem1b) is one of the genes essential for male development and play central roles in sex determination of Caenorhabditis elegans. In this study, we cloned and characterized the full-length Fem1b cDNA from the freshwater prawn Macrobrachium nipponense (MnFem1b) in different tissues and at different developmental stages. Real-time quantitative reverse polymerase chain reaction (RT-qPCR) showed that the MnFem1b gene was expressed in all investigated tissues, with the highest expression level found in the testes. The results revealed that the MnFem1b gene might play roles in aspects of development of the male prawn phenotype. The RT-qPCR also revealed that MnFem1b mRNA expression was significantly increased at 10 days after metamorphosis. The expression levels in all investigated tissues showed a certain degree of sexually dimorphism, the expression levels in males were significantly higher than those in females (P < 0.05). Notably, the highest expression of MnFem1b was found in the testes. The expression of MnFem1b in different tissues indicates that it plays multiple biological functions in M. nipponense.
Assuntos
Síndrome de Resistência a Andrógenos/genética , Proteínas de Artrópodes/genética , Metamorfose Biológica/genética , Palaemonidae/genética , Processos de Determinação Sexual , Sequência de Aminoácidos/genética , Animais , Caenorhabditis elegans/genética , Clonagem Molecular , DNA Complementar/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Masculino , Ovário/crescimento & desenvolvimento , Ovário/metabolismo , Palaemonidae/crescimento & desenvolvimento , Filogenia , RNA Mensageiro/biossíntese , RNA Mensageiro/genéticaRESUMO
Insulin resistance is a key feature of obesity and type 2 diabetes mellitus (T2DM). Interaction of insulin with the insulin receptor (IR) leads to both its auto-phosphorylation and phosphorylation of tyrosine residues on the IR substrate (IRS) proteins, initiating the activation of intracellular signaling cascades. The metabolic effects of IRS are known to be mediated through pathways involving phosphatidyl-inositol 3-kinase (PI-3K), which result in the activation of Akt signaling. The C-terminal region of the IR ectodomain is required to facilitate the conformational changes that are required for high-affinity binding to insulin. Furthermore, the CH2 and CH3 domains in the Fc fragments of immunoglobulins are responsible for their binding to the Fc receptor, which triggers transcytosis. In this study, we created a fusion peptide of the C-terminal end of the human IR ectodomain with the IgG4 Fc fragment, including an intervening polyG fragment to ensure enough space for insulin binding. We named this new peptide "Yiminsu", meaning an insulin sensitizer. The results of our analyses show that Yiminsu significantly facilitates insulin signaling via the activation of Akt in hepatocytes in a dose- and time-dependent manner. Further studies are required to determine whether Yiminsu can act as an insulin sensitizer.
Assuntos
Diabetes Mellitus Tipo 2/tratamento farmacológico , Fragmentos Fab das Imunoglobulinas/farmacologia , Imunoglobulina G/farmacologia , Insulina/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Recombinantes de Fusão/farmacologia , Animais , Células CHO , Linhagem Celular , Clonagem Molecular/métodos , Cricetulus , Diabetes Mellitus Tipo 2/metabolismo , Relação Dose-Resposta a Droga , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Humanos , Fragmentos Fab das Imunoglobulinas/química , Fragmentos Fab das Imunoglobulinas/genética , Imunoglobulina G/química , Imunoglobulina G/genética , Resistência à Insulina , Fosfatidilinositol 3-Quinase/metabolismo , Fosforilação/efeitos dos fármacos , Engenharia de Proteínas , Estrutura Terciária de Proteína , Receptor de Insulina/metabolismo , Receptores Fc/metabolismo , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Transdução de Sinais/efeitos dos fármacos , Tirosina/metabolismoRESUMO
Broad-Complex (BR-C) is an early ecdysone-responsive gene encoding a family of zinc-finger transcription factors. In this study, we isolated the full-length cDNA of a BR-C homolog from the testes of the oriental river prawn (Macrobrachium nipponense), according to established expressed sequence tag information, using the rapid amplification of cDNA ends technique. The homolog was designated as MnBR-C. The full-length cDNA of MnBR-C contained a 1095-bp open reading frame encoding a precursor protein of 365 amino acid residues. Comparative and bioinformatic analyses revealed that MnBR-C exhibited a high degree of homology with BR-C proteins, and contained the BTB and Zf-H2C2-2 domains. Real-time quantitative polymerase chain reaction (qPCR) analysis revealed that the MnBR-C expression level varied significantly in the developing embryo, postembryonic larva, and adult tissue. Real-time qPCR showed that the MnBR-C gene was expressed in all of the tissues investigated, with the highest level of expression in the brain. In addition, MnBR-C was more abundantly expressed in the testes than in the ovaries.
Assuntos
Clonagem Molecular , Regulação da Expressão Gênica no Desenvolvimento , Palaemonidae/genética , Testículo/metabolismo , Fatores de Transcrição/genética , Dedos de Zinco , Sequência de Aminoácidos , Animais , Sequência de Bases , Encéfalo/crescimento & desenvolvimento , Encéfalo/metabolismo , DNA Complementar/genética , DNA Complementar/metabolismo , Embrião não Mamífero , Escherichia coli/genética , Escherichia coli/metabolismo , Feminino , Expressão Gênica , Larva/genética , Larva/crescimento & desenvolvimento , Larva/metabolismo , Masculino , Metamorfose Biológica/genética , Fases de Leitura Aberta , Ovário/crescimento & desenvolvimento , Ovário/metabolismo , Palaemonidae/crescimento & desenvolvimento , Palaemonidae/metabolismo , Estrutura Terciária de Proteína , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Testículo/crescimento & desenvolvimento , Fatores de Transcrição/metabolismoRESUMO
In this study, male-specific lethal 3 homolog (Mnmsl3) was cloned and characterized from the freshwater prawn Macrobrachium nipponense (Crustacea: Decapoda: Palaemonidae) by rapid amplification of cDNA ends. The deduced amino acid sequences of Mnmsl3 showed high-sequence homology to the insect Msl3 and contained a conserved chromatin organization modifier domain and an MORF4-related gene domain. Real-time quantitative reverse transcription-polymerase chain reaction showed that the Mnmsl3 gene was expressed in all the investigated tissues, with the highest level of expression in the testis. The expression level of Mnmsl3 between males and females was different in the gonad (testis or ovary), abdominal ganglion, and heart. The results revealed that the Mnmsl3 gene might play roles in regulating chromatin and in dosage compensation of M. nipponense. Real-time quantitative reverse transcription-polymerase chain reaction also revealed that Mnmsl3 mRNA expression was significantly increased in both 5 and 20 days post-larvae after metamorphosis, suggesting that Mnmsl3 plays complex and important roles in the early embryonic development and sex differentiation of M. nipponense.
Assuntos
Proteínas de Artrópodes/genética , Perfilação da Expressão Gênica , Palaemonidae/genética , Testículo/metabolismo , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/classificação , Sequência de Bases , Clonagem Molecular , DNA Complementar/química , DNA Complementar/genética , Feminino , Cistos Glanglionares/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Larva/genética , Larva/crescimento & desenvolvimento , Masculino , Metamorfose Biológica/genética , Dados de Sequência Molecular , Miocárdio/metabolismo , Ovário/metabolismo , Palaemonidae/embriologia , Palaemonidae/crescimento & desenvolvimento , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Rios , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Fatores SexuaisRESUMO
The gene female sterile homeotic (fsh) plays crucial roles in molecular function, including protein kinase activity and DNA binding, which are involved in biological processes such as terminal region determination and negative regulation of DNA-dependent transcription. Although fsh has been found in Drosophila melanogaster, little is known regarding its expression in crustaceans. In this study, a fsh gene homologue, designated as Mnfsh, was cloned and characterized from the testis of the oriental river prawn, Macrobrachium nipponense, by using EST analysis and the RACE approach for the first time. The full-length cDNA of Mnfsh was 2029 bp, consisting of a 5' UTR of 361 bp, a 3' UTR of 216 bp, and an ORF of 1452 bp encoding 484 amino acids. qRT-PCR analysis showed that the Mnfsh gene was expressed in the testis, ovary, muscle, heart, eyestalk, and abdominal ganglion, with the highest level of expression in the ovary and the lowest in the heart. qRT-PCR analyses showed that the expression levels of Mnfsh mRNA both significantly increased in the zoea stage, the VII larvae, and 1st day post-larvae after metamorphosis. In conclusion, the results of the present study indicate that Mnfsh is an arthropod fsh homologue and probably also plays important roles in embryogenesis, organogenesis, and morphological differentiation of M. nipponense.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento/genética , Palaemonidae/genética , Fatores de Transcrição/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , DNA Complementar/genética , Proteínas de Ligação a DNA/genética , Desenvolvimento Embrionário/genética , Expressão Gênica/genética , Perfilação da Expressão Gênica , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase em Tempo Real , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
This study utilized high-throughput RNA sequencing technology to identify reproduction- and development-related genes of Macrobrachium nipponense by analyzing gene expression profiles of testis and ovary. More than 20 million 1 x 51-bp reads were obtained by Illumina sequencing, generating more than 7.7 and 11.7 million clean reads in the testis and ovary library, respectively. As a result, 10,018 unitags were supposed to be differentially expressed genes (DEGs) between ovary and testis. Compared to the ovary library, 4563 (45.5%) of these DEGs exhibited at least 6-fold upregulated expression, while 5455 (54.5%) DEGs exhibited at least 2-fold downregulated expression in the testis. The Gene Ontology (GO) enrichment analysis showed that 113 GO terms had potential molecular functions in reproduction. The Kyoto Encyclopedia of Genes and Genomes results revealed that the most important pathways may be relevant to reproduction and included 7 pathways. Forty-two genes were identified as reproduction-, development-, and sex-related genes based on GO classification and sequence comparison with other publications, including male reproductive-related LIM protein, spermatogenesis-associated protein, gametocyte-specific factor 1, VASA-like protein, vitellogenin, sex-determining protein fem-1, and other potential candidates. These results will advance research in the field of molecular genetics in M. nipponense and offer a valuable resource for further research related to reproduction in crustaceans.
Assuntos
Ovário/fisiologia , Palaemonidae/genética , Reprodução/genética , Testículo/fisiologia , Animais , Feminino , Perfilação da Expressão Gênica , Ontologia Genética , Sequenciamento de Nucleotídeos em Larga Escala , Masculino , Redes e Vias Metabólicas , Ovário/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Testículo/metabolismoRESUMO
Breast cancer is the most common gynecologic tumor globally that threatens women's health. Lipoic acid is a type of antioxidant that can alleviate oxidative stress damage. Studies showed that lipoic acid could inhibit the proliferation of tumor cells in cervical cancer and colon cancer. This paper intends to explore the combined effect of lipoic acid and paclitaxel on breast cancer cells. Breast cancer MCF-7 cells were divided into four groups: control group, lipoic acid group, paclitaxel group, and a combination group. MTT was applied to detect the drugs' influence on breast cancer cell proliferation. A colony formation test was used to determine the effects on breast cancer cell clone formation rate. Western blot was performed to detect the effects on nuclear factor (NF)-κB. Lipoic acid alone can inhibit tumor cell proliferation and clone formation with time dependence. Compared with the control, paclitaxel alone can significantly suppress tumor cell proliferation and clone formation (P < 0.05). Lipoic acid and paclitaxel in combination obviously strengthened their individual inhibitory effects on tumor cells (P < 0.05). Compared with the paclitaxel alone group, the combination group exhibited more remarkable inhibitory effect (P < 0.05). Lipoic acid alone or combined with paclitaxel can inhibit NF-κB expression and inhibit breast cancer cell proliferation.
Assuntos
Neoplasias da Mama/tratamento farmacológico , NF-kappa B/biossíntese , Paclitaxel/administração & dosagem , Ácido Tióctico/administração & dosagem , Apoptose/efeitos dos fármacos , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Proliferação de Células/efeitos dos fármacos , Feminino , Humanos , Células MCF-7 , NF-kappa B/genéticaRESUMO
The oriental river prawn, Macrobrachium nipponense, is an important aquaculture species in China. The androgenic gland produces hormones that play crucial roles in the differentiation of crustaceans to the male sex. MicroRNA (miRNA) post-transcriptionally regulates many protein-coding genes, influencing important biological and metabolic processes. However, currently, there is no published data identifying miRNA in M. nipponense. In this study, we identified novel miRNA in the androgenic gland of M. nipponense. Using the high-throughput Illumina Solexa system, 1077 miRNA were identified from small RNA libraries by aligning with the de novo androgenic gland transcriptome of M. nipponense (obtained from RNA-Seq) and the sequences in the miRBase21 database. A total of 8,248, 76,011, and 78,307 target genes were predicted in the EST and SRA sequences provided in the NCBI database, and the androgenic gland transcriptome of M. nipponense, respectively. Some potential sex-related miRNA were identified based on the function of the predicted target genes. The results of our study provide new information regarding the miRNA expression in M. nipponense, which could be the basis for further genetic studies on decapod crustaceans.
Assuntos
MicroRNAs/genética , Palaemonidae/genética , Interferência de RNA , RNA Mensageiro/genética , Rios , Animais , Biologia Computacional/métodos , Dosagem de Genes , Regulação da Expressão Gênica , Biblioteca Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Fatores SexuaisRESUMO
MicroRNAs (miRNAs) are small non-coding RNA molecules that play key roles in the regulation of development processes of many tissues and organs at the post-transcriptional level. However, little is known about how they affect chicken gonadal development. We examined the expression of four miRNAs (miR-218, -200b, -196, and -206) in chicken embryonic gonads at embryonic days 3.5-6.5. Their target genes were predicted by miRDB, TargetScan and PicTar algorithms. The expression levels of these four miRNAs differed with sex to varying degrees; miR-200b was expressed at a significantly higher level in female gonads during the entire interval. The whole mount in situ hybridization result showed considerably higher expression of miR-200b in females than in males in E5.5 embryos. The miRNA target scanning results indicated several genes with functions in gonad development and gonad function. We conclude that miR-200b is involved in the regulation of gonad development and sexual differentiation of chicken embryos.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento , MicroRNAs/biossíntese , Diferenciação Sexual/genética , Animais , Embrião de Galinha , Galinhas/genética , Feminino , Perfilação da Expressão Gênica , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Hibridização In Situ , Masculino , MicroRNAs/genética , Ovário/crescimento & desenvolvimento , Ovário/metabolismoRESUMO
A genome-wide association study revealed that a single nucleotide polymorphism, CLPTM1L - rs401681 (G>A), located at the 5p15.33 locus was significantly associated with increased risk of various cancers; however, its association with lung cancer is currently inconclusive. In order to explore the relationship between this polymorphism and lung cancer risk more precisely, we performed a meta-analysis of eight eligible studies involving 9935 cases and 11,261 controls. The pooled odds ratio (OR) and the 95% confidence interval (CI) were calculated using a fixed- or random-effect models. Results indicated that this polymorphism was significantly associated with lung cancer risk in all genetic models (GA vs GG: OR = 0.88, 95%CI = 0.83-0.94; AA vs GG: OR = 0.81, 95%CI = 0.70-0.93; AA/GA vs GG: OR = 0.86, 95%CI = 0.81-0.91; AA vs GA/GG: OR = 0.86, 95%CI = 0.76-0.99). An analysis stratified by ethnicity and source of controls revealed a significantly decreased risk among European groups and population-based studies in all genetic models, and among Asian populations only in the dominant model comparison. Additionally, in a subgroup analysis by histology type, the CLPTM1L rs401681 polymorphism was found to significantly decrease the risks of both adenocarcinoma and squamous cell carcinoma of the lung in all genetic models. In conclusion, our study indicated that the CLPTM1L - rs401681 (G>A) polymorphism was significantly associated with decreased lung cancer risk, especially among European populations. Due to some minor limitations, our findings should be confirmed in further studies.
Assuntos
Predisposição Genética para Doença , Neoplasias Pulmonares/genética , Proteínas de Membrana/genética , Proteínas de Neoplasias/genética , Polimorfismo de Nucleotídeo Único , Alelos , Estudos de Casos e Controles , Genótipo , Humanos , Neoplasias Pulmonares/epidemiologia , Razão de Chances , Viés de Publicação , RiscoRESUMO
Increased disease resistance through improved general immune capacity would be beneficial for the welfare and productivity of farm animals. Classical swine fever (CSF) is a contagious disease in farm animals. The immunoglobulin G (IgG) blocking percentage of CSF virus (CSFV) in serum is an essential diagnostic parameter in veterinary practice. In addition, lysozymes are a part of the innate immune system. To identify quantitative trait loci (QTL) for IgG blocking percentage of CSFV and lysozyme concentration, IgG blocking percentage and lysozyme concentration in serum were measured in a composite pig population before and after challenge with modified live CSF vaccine. Through genome-wide mapping by MQREML analysis and the SOLAR software, several QTL for the lysozyme concentration and the IgG blocking percentage of CSFV were identified, respectively. Within these QTL regions, some known genes were revealed, and some of them may serve as candidate genes in the pig.
Assuntos
Anticorpos Antivirais/genética , Vírus da Febre Suína Clássica/imunologia , Imunoglobulina G/genética , Muramidase/genética , Locos de Características Quantitativas , Imunidade Adaptativa/genética , Animais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Mapeamento Cromossômico , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Muramidase/sangue , SuínosRESUMO
PURPOSE: T cells are dominant in the immune regulation of malignant pleural effusion (MPE). However, it is unclear about the role of IL-17+ T cells, particularly for IL-17+CD8+ Tc17 cells in antitumor immunity. This retrospective study is aimed at evaluating the prognostic significance of IL-17+ T cells in patients with MPE. METHODS: The frequency of IL-17+CD4+ Th17 and IL-17+CD8+ Tc17 cells in peripheral blood (PB), pleural fluids (PF), and tumor tissues in 24 patients undergoing thoracoscopy was determined by flow cytometry, immunohistochemistry, and ELISA. The association among the different measures was analyzed by Spearman's correlation tests. RESULTS: The percentages of PF Th17 and Tc17 cells were significantly higher than those in the PB of MPE patients and healthy controls (p < 0.01). Analysis of Th17 and Tc17 cells in the tumor tissues indicated that the percentages of Th17 and Tc17 cells in the invading tumor edge were significantly higher than those in the non-tumor tissues and intra-tumor regions (p < 0.05). More importantly, the percentages of IL-17+ T cells were associated with prolonged survival of patients with MPE. CONCLUSIONS: Both Th17 and Tc17 cells were involved in the tumor immunity against MPE. Increased frequency of Tc17 cells may serve as a biomarker for the prognosis of patients with MPE.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Interleucina-17/biossíntese , Derrame Pleural Maligno/imunologia , Subpopulações de Linfócitos T/imunologia , Células Th17/imunologia , Idoso , Idoso de 80 Anos ou mais , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
Gastric cancer is a major health problem worldwide; it is the second most common cause of cancer death in the world. Recent studies indicate that the high-mobility group (HMG) of chromosomal proteins is associated with cancer progression. However, HMGB3 has been little studied. We analyzed the co-expression network between HMGB3 and differentially-expressed genes in the GSE17187 database, identifying the relevant transcription factors, and the conserved domain of HMGB3 to understand the underlying regulation mechanisms involved in gastric cancer. Thirty-one relationships between 11 differentially-expressed genes were included in a co-expression network; many of these genes have been identified as related to cancer, including TBX5 and TFR2. Further analysis identified nine transcription factors, these being GATA3, MZF1, GATA1, GATA2, SRY, REL, NFYB, NFYC, and NFYA, which could interact with HMGB3 to regulate target gene expression and consequently regulate gastric cancer cell proliferation, migration and invasion. The HMG-box domain was very similar in various species, with only a few amino acid changes, indicating conserved functions in HMG-box. This information helps to provide insight into the molecular mechanisms of HMGB3 in human gastric cancer.
Assuntos
Regulação Neoplásica da Expressão Gênica , Proteína HMGB3/genética , Neoplasias Gástricas/genética , Sequência de Aminoácidos , Sequência Conservada , Proteína HMGB3/química , Humanos , Dados de Sequência Molecular , Estrutura Terciária de Proteína , Neoplasias Gástricas/metabolismo , TranscriptomaRESUMO
Mutations in the V-raf murine sarcoma viral oncogene homolog B1 gene (BRAF) play an important role in the pathogenesis of papillary thyroid cancer (PTC). In this study, a BRAF V600E mutation was detected in formalin-fixed and paraffin-embedded PTC samples using multiplex allele-specific polymerase chain reaction and denaturing high-performance liquid chromatography. The sensitivity of the assays was validated in two cell lines, K1 and RO82-W-1, which were respectively positive and negative for the mutation. The method enabled detection of very low concentrations (~1%) of the mutation, with positive findings obtained in 119 of the 187 samples (63.6%). The mutation was not detected in 20 cases of benign thyroid diseases. The presence of the mutation was significantly associated with ages >45 years, the tumor-nodes-metastasis stage, region of metastasis, and clinical outcome (P < 0.05). Although no correlation was found between the mutation and lymph node metastasis, the proportions of patients carrying a mutation differed significantly between central and lateral cervical lymph node metastasis (P < 0.05). Multivariate logistic regression analysis also indicated the presence of mutation, tumor size, and cervical lymph node metastasis to be independent predictors for recurrence and distant metastasis. We conclude that a high proportion of PTC cases likely harbors the BRAF V600E mutation. This mutation can be used as an independent factor for predicting the recurrence and distal metastasis of PTC tumors.
Assuntos
Carcinoma/diagnóstico , Carcinoma/genética , Mutação , Proteínas Proto-Oncogênicas B-raf/genética , Neoplasias da Glândula Tireoide/diagnóstico , Neoplasias da Glândula Tireoide/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Alelos , Substituição de Aminoácidos , Carcinoma Papilar , Criança , Cromatografia Líquida de Alta Pressão , Análise Mutacional de DNA , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Multiplex , Estadiamento de Neoplasias , Reprodutibilidade dos Testes , Câncer Papilífero da Tireoide , Adulto JovemRESUMO
To assess the genetic status of this species, the genetic diversity of wild Macrobrachium nipponense from seven geographic locations in the Yellow River basin were investigated using 20 polymorphic microsatellite DNA loci. The genetic diversity between populations was indicated by the mean number of alleles per locus and mean observed heterozygosity (H) and the expected H, which was arranged from 2 to 10, from 0.4705 to 0.5731, and from 0.5174 to 0.6146, respectively. Hardy-Weinberg equilibrium analysis indicated that a deficiency of heterozygotes existed in all seven populations. Both the F(ST) and AMOVA analyses showed that there is significant difference on population differentiation among populations. The UPGMA clustering tree demonstrated that their close relationship is consistent with their geographic proximity. The data suggest that this Yellow River population has a wide genetic base that is suitable for breeding.
Assuntos
Repetições de Microssatélites , Palaemonidae/genética , Polimorfismo Genético , Alelos , Animais , Marcadores Genéticos , Heterozigoto , MutaçãoRESUMO
In this study, two Sxl gene homologs, designated as Mnsxl1 and Mnsxl2, were cloned and characterized from the freshwater prawn Macrobrachium nipponense by rapid amplification of cDNA ends. The deduced amino acid sequences of Mnsxl1 and Mnsxl2 showed high sequence homology to the insect Sxl and contained conserved domains in two RNA-binding motifs. Real-time quantitative reverse transcription-polymerase chain reaction (RT-QPCR) showed that the Mnsxl1 and Mnsxl2 genes were expressed in all investigated tissues, with the highest level of expression in the intestine and liver. RT-QPCR also revealed that Mnsxl1 and Mnsxl2 mRNAs expressions were both significantly increased at 5 and 20 days post-larvae after metamorphosis. Thus, the results of the present study imply that Mnsxl1 and Mnsxl2 play complex and important roles in the sex differentiation of M. nipponense.
Assuntos
Proteínas de Artrópodes/genética , Palaemonidae/genética , Proteínas de Ligação a RNA/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Larva/genética , Larva/metabolismo , Masculino , Metamorfose Biológica , Dados de Sequência Molecular , Especificidade de Órgãos , Palaemonidae/crescimento & desenvolvimento , Palaemonidae/metabolismo , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Processos de Determinação Sexual , TranscriptomaRESUMO
The STAM protein plays an important role in the cytokine-related JAK/STAT pathway. We selected the STAM2 gene as a candidate gene that could be linked to growth performance in analysis of a Chinese cattle breed (Wuchuan Black cattle). We examined genetic variants in the promoter region of the STAM2 gene and their associations with eight growth traits in 159 individuals. Seven SNPs, which included six new SNPs for the SNP database, were found. The core promoter region was identified with a bioinformatic software. This analysis also showed that the SNPs have a significant influence on the function and structure of the STAM2 promoter in terms of RNA secondary structure, CpG island, and transcription factor binding sites. Association analysis demonstrated that G-102A is significantly associated with withers height, heart girth, cannon circumference, chest width, and hip height in this population, which leads us to suggest that G-102A is a useful SNP marker for cattle growth performance. Animals with the genotype AA had higher mean values for withers height, cannon circumference, chest width, and hip height than those with GG and AG genotypes. This SNP of the STAM2 gene could be applied in marker-assisted selection for improving growth performance in cattle.