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1.
Int J Biol Macromol ; 149: 1034-1041, 2020 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-32027900

RESUMO

Chitooligosaccharides (COS) are hydrolytic products of chitosan that are essential in functional food, medicine, and other fields due to their biological activities. Commercial COS are often prepared by the hydrolysis of chitosan by chitosanase. In this study, a glycoside hydrolase family 46 cluster B chitosanase from Bacillus amyloliquefaciens (BaCsn46B) was efficiently expressed in Pichia pastoris. The recombinant enzyme was secreted into the culture medium that reached a total extracellular protein concentration of 4.5 g/L with an activity of 8907.2 U/mL in a high cell density fermenter (5 L). The molecular mass of deglycosylated BaCsn46B was 29.0 kDa. Purified BaCsn46B exhibited excellent enzymatic properties, which had high specific activity (2380.5 U/mg) under optimal reaction conditions (55 °C and pH 6.5). BaCsn46B hydrolyzed chitosan yielded a series of COS with different degrees of polymerization by endo-type cleavage. The end hydrolytic products of BaCsn46B were chitobiose and chitotriose, while no monosaccharide yield was evident in the hydrolytic reaction. The excellent secreted expression level and hydrolytic performance make the enzyme a desirable biocatalyst for the industrial preparation of COS.

2.
Genomics ; 2020 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-32045669

RESUMO

In this study, we determined the complete mitogenome sequence of Calappa bilineata, which is the first mitogenome of Calappidae up to now. The total length is 15,606 bp and includes 13 protein-coding genes, 22 transfer RNAs, two ribosomal RNAs and one control region. The genome composition is highly A + T biased (68.7%), and exhibits a negative AT-skew (-0.010) and GC-skew (-0.267). As with other invertebrate mitogenomes, the PCGs start with the standard ATN and stop with the standard TAN codons or incomplete T. Phylogenetic analysis showed that C. bilineata was most closely related to Matuta planipes (Matutidae), and these two species formed a sister clade, constituting a Calappoidea group and forming a sister clade with part of Eriphioidea. The existence of the polyphyletic families raised doubts over the traditional classification system. These results will help to better understand the features of the C. bilineata mitogenome and lay foundation for further evolutionary relationships within Brachyura.

3.
Int J Biol Macromol ; 2020 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-32032708

RESUMO

Mitochondrial genome (mitogenome) structure and gene order are generally considered conserved in vertebrates. However, the flatfish (Pleuronectiformes) mitogenomes exhibit the most diversified gene rearrangement patterns. Here, we report a newly sequenced mitogenome of Cynoglossus melampetalus (Pleuronectiformes: Cynoglossidae). The total length of the C. melampetalus mitogenome is 16,651 bp, containing 13 protein-coding genes, two ribosomal RNAs, 22 transfer RNAs, a putative control region, and an L-strand replication origin. Like all previously reported tongue sole (Cynoglossinae) mitogenomes, the C. melampetalus tRNA-Gln gene is inverted from the light to the heavy strand (Q inversion), accompanied by the translocation of CR, which is downstream to the 3'-end of ND1. In addition, we observed a unique tRNA-Ile-Met-Glu (IMQ) gene order that differed from the tRNA-Glu-Ile-Met (QIM) order previously reported for other 14 Cynoglossinae mitogenomes. To our knowledge, it is the first report of two different patterns of mitogenomic gene-arrangement within the same genus in teleost. According to the Q inversion, Met pseudogene (ψMet) and long intergenic gap (186 bp) between M and Q genes, the observed gene rearrangement pattern were presumably supported by mitochondrial recombination and tandem duplication/random loss models. The reduced trend of the intergenic gap between Q and I also suggests that the event of gene rearrangement can be traced back to early Cynoglossinae differentiation.

4.
Medicine (Baltimore) ; 99(4): e18887, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31977897

RESUMO

INTRODUCTION: MYH9-related disease (MYH9-RD) is a rare autosomal dominant disorder caused by mutations in MYH9, which is responsible for encoding nonmuscle myosin heavy chains IIA (NMMHCIIA). MYH9-RD is clinically characterized by congenital macrothrombocytopenia, granulocyte inclusions variably associated with the risk of developing progressive sensorineural deafness, cataracts and nephropathy. PATIENT CONCERNS: A 5-year-old boy had a history of a mild bleeding tendency and chronic thrombocytopenia, first identified at four months of age. No other family members were noted to have similar clinical features or hematologic disorders. DIAGNOSES: The boy was diagnosed with MYH9-RD. Light microscopic examination of peripheral blood films (Wright-Giemsa stain) showed marked platelet macrocytosis with giant platelets and basophilic Döhle-like inclusions in 83% of the neutrophils. Immunofluorescence analysis disclosed a type II pattern, manifested by neutrophils which contained several circle-to-oval shaped cytoplasmic NMMMHCA-positive granules. Sequencing analysis of MYH9-RD genes was carried out and revealed a novel missense mutation of c.97T>G (p.W33G) in the patient but not in his parents. INTERVENTION: No treatment is necessary. Recognition of MYH9-RD is important to Avoiding unnecessary and potentially harmful treatments. OUTCOMES: The patient's condition remained stable during the follow-up. CONCLUSIONS: As a result of identifying this missense mutation in this particular case, we have added c.97T>G (p.W33G) to the broad spectrum of potential MYH9 mutations.


Assuntos
Perda Auditiva Neurossensorial/genética , Trombocitopenia/congênito , Pré-Escolar , Imunofluorescência , Perda Auditiva Neurossensorial/diagnóstico , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Mutação de Sentido Incorreto , Neutrófilos/patologia , Trombocitopenia/diagnóstico , Trombocitopenia/genética
5.
Carbohydr Polym ; 230: 115674, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31887864

RESUMO

A novel macromolecular polysaccharide VGPⅠ-a was purified from Volvariella volvacea fruiting bodies with ultrasound-assisted extraction, ion exchange and gel chromatography. VGPⅠ-a was proved as a α- glucan with Mw of 1435.6 kDa and contained a 1,4-linked d-Glcp backbone with the substitution at C-6 with 1-linked d-Glcp residue. Congo-red test, AFM and SEM analysis showed VGPⅠ-a had a triple-helical conformation and the interacted chains to form a small screw-rod and dispersed appearance. VGPⅠ-a had no cytotoxic effect on macrophage RAW264.7 cells in vitro and significantly enhanced the production and mRNA expression of NO, TNF-α, IL-6 and IL-1ß in a dose-dependent manner. Further analyses demonstrated that VGPⅠ-a activated the MAPK signaling pathway by improving the phosphorylated levels of p38, JNK and ERK in RAW264.7 cells to promote the expression and secretion of above cytokines. These findings would provide a better understanding of V. volvacea glucan and its potential immunomodulating mechanisms.

6.
Anim Reprod Sci ; 211: 106231, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31785644

RESUMO

Estrogen related receptors (ERRs) are widely detected in vertebrates and apparently have functions in reproduction. The functions of ERRs in reproduction of invertebrates, especially in mollusk cephalopods, are largely unknown. In the present study, An homologue of vertebrate ERR gene was first cloned from female Sepiella japonica, an important Cephalopod species in coastal water of China. Results indicate the S. japonica ERR (sjERR) gene is comprised of 1513 nucleotides, containing a 1389 bp open reading frame, which encode for 463 amino acid (aa) residues. The deduced sjERR protein possessed six typical nuclear receptors (NR) domains (A-F), with a DNA-binding domain (DBD) and a highly conserved ligand-binding domain (LBD), compared to the other molluscan ERRs. Results from tissue analyses indicated that sjERR mRNA transcript abundance was in largest amounts in tissues of the brain, liver, ovary that are possibly involved in reproduction. The sjERR mRNA transcript abundance was temporally regulated during the different sexual maturation phases of female S. japonica and was affected by in vivo administrations of vertebrate steroid estradiol-17ß (E2). An in vivo knockdown of sjERR gene expression resulted in a marked down-regulation in expression of genes involved in ovarian development, such as Vitellogenin, CDK1, and Cyclin B, indicating there is a possible involvement of sjERR in reproduction. Both fusion protein transient transfections and immunohistochemical analyses indicated a presence of sjERR in the nucleus, implying a possible mechanism of action of the sjERR in the nucleus through activation of specific gene transcriptions.

7.
Infect Drug Resist ; 12: 3653-3661, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31819548

RESUMO

Purpose: This study aimed to investigate the species distributions and drug sensitivities among 19 strains of Nocardia isolated from Yantai, China, from 2017 to 2019. Patients and methods: Definitive species identification was performed by sequencing a fragment of the 16S rRNA gene (1480 bp) and by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). The susceptibilities of the isolates to 15 commonly-used antibiotics were tested using the microbroth dilution method. Results: Among the 19 Nocardia isolates, five species were confirmed. Seventeen of the 19 Nocardia spp. strains were identified consistently by the two methods, while two isolates of N. cyriacigeorgica were misidentified as N. otitidiscaviarum by MALDI-TOF MS. N. farcinica was the most common species (8/19), followed by N. cyriacigeorgica (6/19), N. otitidiscaviarum (2/19), N. brasiliensis (2/19), and N. nova (1/19). All isolates were susceptible to trimethoprim-sulfamethoxazole and amikacin, followed by linezolid and tigecycline (94.7% susceptibility rates). The sensitivity and minimum inhibitory concentration patterns for ciprofloxacin, moxifloxacin, clarithromycin, and tobramycin were significantly correlated with the species. Conclusion: These results regarding the distribution and antibiotic resistance features of Nocardia species further our understanding of the diversity of Nocardia species circulating in Yantai, China, and thus support the use of more accurate empirical treatments.

8.
Molecules ; 24(22)2019 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-31703405

RESUMO

A simple, sensitive and effective magnetic solid-phase extraction (MSPE) technique was developed for the extraction of pyrethroid pesticides from environmental water samples, followed by gas chromatography tandem triple quadrupole mass spectrometry determination. An adsorbent of magnetic zeolitic imidazolate framework-8@deep eutectic solvent (M-ZIF-8@DES) was prepared using deep eutectic solvent coated on the surface of M-ZIF-8. The features of M-ZIF-8@DES were confirmed by material characterizations, and the results indicated that M-ZIF-8@DES has a good magnetism (61.3 emu g-1), a decent surface area (96.83 m2 g-1) and pore volume (0.292 mL g-1). Single factor experiments were carried out to investigate the effect of different conditions on the performance of MSPE. Under the optimal conditions, the developed method performs good linearity (R2 ≥ 0.9916) in the concentration range of 1-500 µg L-1. The limits of detection were in the range of 0.05-0.21 µg L-1 (signal/noise = 3/1). The intraday relative standard deviation (RSD) and interday RSD were less than 9.40%. Finally, the proposed technique was applied for the determination of pyrethroid pesticides in environmental water samples. This work shows the potential of DES-modified metal-organic frameworks for different sample pretreatment techniques.

9.
Medicine (Baltimore) ; 98(48): e18121, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31770241

RESUMO

RATIONALE: Congenital generalized lipodystrophy (CGL) is a rare autosomal recessive hereditary disease. It is associated with metabolic complications and epilepsy is rare. PATIENT CONCERNS AND DIAGNOSES: One child with BSCL2 mutation and CGL accompanied by progressive myoclonic epilepsyDiagnosis: He was diagnosed with epilepsy, CGL, and severe malnutrition. INTERVENTIONS: He was treated with sodium valproate, baclofen, aripiprazole, benzhexol, and lamotrigine for epilepsy. OUTCOMES: After 16 days of medical treatment for epilepsy, the disease was improved and the child was discharged with gastric tube inserted for the management of malnutrition. LESSONS: CGL and progressive myoclonic epilepsy is rare, and the epilepsy is partially refractory to treatments. In this particular case, the nutritional status was compromised as a complication of progressive myoclonic epilepsy and had to be managed.


Assuntos
Lipodistrofia Generalizada Congênita/complicações , Epilepsias Mioclônicas Progressivas/congênito , Criança , Humanos , Masculino
10.
Genomics ; 2019 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-31655177

RESUMO

The complete mitochondrial genomes (mitogenomes) can indicate phylogenetic relationships among organisms, as well as useful information about the process of molecular evolution and gene rearrangement mechanisms. However, knowledge on the complete mitogenome of Coenobitidae (Decapoda: Anomura) is quite scarce. Here, we describe in detail the complete mitogenome of Coenobita brevimanus, which is 16,393 bp in length, and contains 13 protein-coding genes, two ribosomal RNA, 22 transfer RNA genes, as well as a putative control region. The genome composition shows a moderate A + T bias (65.0%), and exhibited a negative AT-skew (-0.148) and a positive GC-skew (0.183). Five gene clusters (or genes) involving eleven tRNAs and two PCGs were found to have rearranged with respect to the pancrustacean ground pattern gene order. Duplication-random loss and recombination models were determined as most likely to explain the observed large-scale gene rearrangements. Phylogenetic analysis placed all Coenobitidae species into one clade. The polyphyly of Paguroidea was well supported, whereas the non-monophyly of Galatheoidea was inconsistence with previous findings on Anomura. Taken together, our results help to better understand gene rearrangement process and the evolutionary status of C. brevimanus and lay a foundation for further phylogenetic studies of Anomura.

11.
Cell Host Microbe ; 26(4): 551-563.e6, 2019 10 09.
Artigo em Inglês | MEDLINE | ID: mdl-31540829

RESUMO

During infection, Legionella pneumophila translocates over 300 effector proteins into the host cytosol, allowing the pathogen to establish an endoplasmic reticulum (ER)-like Legionella-containing vacuole (LCV) that supports bacterial replication. Here, we perform a genome-wide CRISPR-Cas9 screen and secondary targeted screens in U937 human monocyte/macrophage-like cells to systematically identify host factors that regulate killing by L. pneumophila. The screens reveal known host factors hijacked by L. pneumophila, as well as genes spanning diverse trafficking and signaling pathways previously not linked to L. pneumophila pathogenesis. We further characterize C1orf43 and KIAA1109 as regulators of phagocytosis and show that RAB10 and its chaperone RABIF are required for optimal L. pneumophila replication and ER recruitment to the LCV. Finally, we show that Rab10 protein is recruited to the LCV and ubiquitinated by the effectors SidC/SdcA. Collectively, our results provide a wealth of previously undescribed insights into L. pneumophila pathogenesis and mammalian cell function.

12.
Fish Shellfish Immunol ; 94: 634-642, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31533082

RESUMO

Long non-coding RNA refers to an RNA transcript of a non-coding protein with a sequence length greater than 200 bp. More and more reports indicated that lncRNA was involved in the regulation of gene expression as a signalling molecule, an inducing molecule, a leader molecule and a scaffold molecule. Previous studies have sequenced the draft genome and several transcriptome data sets for protein-coding genes of the large yellow croaker (Larimichthys crocea), but little is known about the expression and function of lncRNAs in this species. In order to obtain a catalogue of lncRNAs for this croaker, Vibrio parahaemolyticus infection challenge experiment was conducted and long non-coding RNA sequences were obtained. Using high-throughput sequencing of lncRNA, a total of 73,233 high-confidence transcripts were reconstructed in 32,726 loci, recovering most of the expressed reference transcripts, and 6473 novel expressed loci were identified. The tissue expression profile revealed that most lacunas were specifically enriched in distinct tissues. A set of 163 lncRNAs were identified as being specifically expressed in the spleen and may be involved in the immune response. It is the first time to identify specific lncRNAs in the L. crocea systematically in this croaker, aiming to benefit the future genomic study of this species.

13.
Anim Reprod Sci ; 208: 106107, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31405483

RESUMO

The membrane progestin receptor (mPR) family has been characterized in several species, including fish, frogs, rats, and humans. Results of previous studies indicate mPRs mediate the rapid, nongenomic action of progestins. In this study, the full-length cDNA of Sepiella japonica mPR-beta (mPRß) and mPR-gamma (mPRγ) were characterized. Furthermore, sjmPR mRNA relative abundances were assessed for different tissues. There was also determination of the subcellular localization of mPRs, and investigation of the effect of sjmPRs on ovarian development via proximate actions on the brain and ovary of S. japonica. Results of tissue distribution assays indicated mPRß and mPRγ transcripts were present predominantly in the brain and ovary. As ovaries developed, the abundance of SjmPRs mRNA transcripts increased and peaked during the interstitial growth phase (III), followed by a marked decrease afterward in both the brain and ovary. In addition, confocal microscopy evaluations of HEK293 T cells expressing the mPRs-EGFP gene indicated both SjmPRß and SjmPRγ were localized in the plasma membrane of HEK293 T cells. Taken together, these findings indicate S. japonica protein is a membrane progestin receptor capable of inducing ovary maturation in cephalopods.


Assuntos
Decapodiformes/metabolismo , Regulação da Expressão Gênica/fisiologia , RNA Mensageiro/metabolismo , Receptores de Progesterona/metabolismo , Animais , Feminino , Masculino , RNA Mensageiro/genética
14.
Front Neurosci ; 13: 831, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31440132

RESUMO

ß-Catenin is a key component of the canonical Wnt signaling pathway. It has been shown to have an important role in formation of the neuromuscular junction. Our previous studies showed that in the absence of ß-catenin, the resting membrane potential (RMP) is depolarized in muscle cells and expression of the α2 subunit of sodium/potassium adenosine triphosphatase (α2NKA) is reduced. To understand the underlying mechanisms, we investigated the electrophysiologic properties of a primary cell line derived from mouse myoblasts (C2C12 cells) that were transfected with small-interfering RNAs and over-expressed plasmids targeting ß-catenin. We found that the RMP was depolarized in ß-catenin knocked-down C2C12 cells and was unchanged in ß-catenin over-expressed muscle cells. An action potential (AP) was not released by knockdown or over-expression of ß-catenin. α2NKA expression was reduced by ß-catenin knockdown, and increased by ß-catenin over-expression. We showed that ß-catenin could interact physically with α2NKA (but not with α1NKA) in muscle cells. NKA activity and α2NKA content in the cell membranes of skeletal muscle cells were modulated positively by ß-catenin. These results suggested that ß-catenin (at least in part) regulates the RMP and AP in muscle cells, and does so by regulating α2NKA.

15.
Mol Ther Nucleic Acids ; 17: 405-413, 2019 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-31310946

RESUMO

Lung cancer is the most common cancer all around the world, with high morbidity and mortality. Long noncoding RNA (lncRNA) has been reported to have a critical role in non-small-cell lung cancer (NSCLC) proliferation and migration. In the present study, we analyzed The Cancer Genome Atlas (TCGA) data, and we found that lncRNA Small Nucleolar RNA Host Gene 17 (SNHG17) was upregulated in NSCLC driven by the amplification of copy number, indicating the special role of SNHG17 in NSCLC. The full exact length of SNHG17 was determined by rapid amplification of cDNA ends (RACE). We modulated SNHG17 expression by RNAi and a series of functional assays were performed. Flow cytometry was used to explore the involvement of SNHG17 in NSCLC cell apoptosis. Results showed that the knockdown of SNHG17 inhibited the proliferation and migration and promoted the apoptosis of NSCLC cells. We acquired the global gene expression profile regulated by SNHG17 in A549 through RNA sequencing (RNA-seq) assays. We found 637 genes were upregulated while 581 genes were downregulated. We selected three genes (FOXA1, XAF1, and BIK) that were closely related to proliferation and apoptosis, and we confirmed their altered expression in A549 and PC-9 cells treated with small interfering RNA si-SNHG17. Our findings indicated gene amplification-driven lncRNA SNHG17 promotes cell proliferation and migration in NSCLC, suggesting its potential value as a biomarker in NSCLC.

16.
Cell Cycle ; 18(16): 1893-1907, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31208297

RESUMO

The role of inflammation response has been well documented in the development of acute lung injury (ALI). However, little is known about the functions of miRNAs in the regulation of inflammation in ALI. The aim of this study was to explore the effects of miRNAs in the regulation of inflammation in ALI and to elucidate the biomolecular mechanisms responsible for these effects. The expression profiles of miRNAs in lung tissues from lipopolysaccharide (LPS)-induced ALI mice model were analyzed using a microarray. It was observed that microRNA-221-3p (miR-221) was significantly increased in lung tissues in ALI mice. The inhibition of miR-221 attenuated lung injury including decreased lung W/D weight ratio and lung permeability and survival rates of ALI mice, as well as apoptosis, whereas its agomir-mediated upregulation exacerbated the lung injury. Concomitantly, miR-221 inhibition significantly reduced LPS-induced pulmonary inflammation, while LPS-induced pulmonary inflammation was aggravated by miR-221 upregulation. Of note, suppressor of cytokine signaling-1 (SOCS1), an effective suppressor of the NF-κB signaling pathway, was found to be a direct target of miR-221 in RAW264.7 cells. Overexpression of SOCS1 by pcDNA-SOCS1 plasmids markedly reversed the miR-221 inhibition-mediated inhibitory effects on inflammation and apoptosis in LPS-treated RAW264.7 cells. Finally, it was found that miR-221 inhibition suppressed LPS induced the activation of the NF-κB signaling pathway, as demonstrated by downregulation of phosphorylated-IκBα, p-p65 and upregulation of IκBα, whilst miR-221 overexpression had an opposite result in ALI mice. Our findings demonstrate that inhibition of miR-221 can alleviate LPS-induced inflammation via inactivation of SOCS1/NF-κB signaling pathway in ALI mice.

17.
Int J Biol Macromol ; 135: 609-618, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31132441

RESUMO

Generally, a teleostean group possesses only one type or a set of similar mitochondrial gene arrangement. However, two types of gene arrangement have been identified in the mitochondrial genomes (mitogenomes) of Anguilliformes. Here, a newly sequenced mitogenome of Ophichthus brevicaudatus (Anguilliformes; Ophichthidae) was presented. The total length of the O. brevicaudatus mitogenome was 17,773 bp, and it contained 13 protein-coding genes (PCGs), two ribosomal RNAs (rRNAs), 22 transfer RNA (tRNA) genes, and two identical control regions (CRs). The gene order differed from that of the typical vertebrate mitogenomes. The genes ND6 and the conjoint trnE were translocated to the location between trnT and trnP, and one of the duplicated CR was translocated to the upstream of the ND6. The duplication-random loss model was adopted to explain the gene rearrangement events in this mitogenome. The most comprehensive phylogenetic trees of Anguilliformes based on complete mitogenome was constructed. The non-monophyly of Congridae was well supported, whereas the non-monophyly of Derichthyidae and Chlopsidae was not supported. These results provide insight into gene arrangement features of anguilliform mitogenomes and lay the foundation for further phylogenetic studies on Anguilliformes.


Assuntos
Enguias/genética , Ordem dos Genes/genética , Genoma Mitocondrial/genética , Filogenia , Animais , Códon/genética , Genômica
18.
Int J Mol Med ; 44(1): 346-356, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31115501

RESUMO

Numerous microRNAs (miRs) are dysregulated in non­small cell lung cancer (NSCLC), serving pivotal roles in its formation and progression. miR­625 is dysregulated in several types of human cancer, but its involvement in the formation and development of NSCLC remains poorly understood. In the present study, we aimed to investigate miR­625 expression in NSCLC and its role in regulating NSCLC cell behavior. miR­625 expression in NSCLC tissues and cell lines was detected using reverse transcription­quantitative polymerase chain reaction. The effects of miR­625 overexpression on NSCLC cell proliferation, apoptosis, migration and invasion in vitro were assessed using an MTT assay, flow cytometry, and cell migration and invasion assays, respectively. The effects of miR­625 upregulation on NSCLC growth were evaluated in an in vivo xenograft model. The molecular mechanisms underlying the tumor­suppressing roles of miR­625 in NSCLC were explored in detail. miR­625 expression was determined to be downregulated in NSCLC tissues and cell lines. This decreased expression was associated with advanced clinical features and poor overall survival of patients with NSCLC. Exogenous miR­625 expression suppressed NSCLC cell proliferation, migration and invasion, and induced apoptosis in vitro. miR­625 upregulation hindered NSCLC tumor growth in vivo. Homeobox B5 (HOXB5) was proposed to be the direct target gene of miR­625 in NSCLC cells. The tumor­suppressing effects of HOXB5 silencing were similar to those of miR­625 overexpression in NSCLC cells. In rescue experiments, HOXB5 overexpression partially reversed the inhibitory effects of miR­625 in NSCLC cells. miR­625 upregulation directly targeted HOXB5 to deactivate the Wnt/ß­catenin signaling pathway in NSCLC cells in vitro and in vivo. miR­625 was determined to be associated with HOXB5 suppression and Wnt/ß­catenin pathway deactivation, which in turn inhibited the aggressive behavior of NSCLC cells in vitro and in vivo.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/metabolismo , Proteínas de Homeodomínio/metabolismo , Neoplasias Pulmonares/metabolismo , MicroRNAs/metabolismo , Proteínas de Neoplasias/metabolismo , RNA Neoplásico/metabolismo , Via de Sinalização Wnt , Células A549 , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Movimento Celular/genética , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Proteínas de Homeodomínio/genética , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , MicroRNAs/genética , Invasividade Neoplásica , Proteínas de Neoplasias/genética , RNA Neoplásico/genética , beta Catenina/genética , beta Catenina/metabolismo
19.
Dev Comp Immunol ; 98: 108-118, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31051196

RESUMO

TAK1-binding proteins (TABs) are important immune protein involved in various intracellular signalling pathways. Here, TAB1-3 (lcTAB1-3) were characterized from Larimichthys crocea. The predicted 1524 bp coding sequence of lcTAB1 encoded a 507-residue protein, while lcTAB2 (2271 bp) and lcTAB3 (1836 bp) encoded 756 and 611 residue proteins, respectively. Their sequence shared conserved domain structures and functional sites with their orthologs from other species. The expression of lcTAB1-3 were detected in all tested tissues, which were upregulated in spleen, liver and kidney following Vibrio parahemolyticus infection. Immunofluorescence staining revealed that lcTAB1 were localized in cytoplasm, while lcTAB2 and lcTAB3 were in the endsome. Moreover, the NF-κB protein level was obviously upregulated after the co-overexpression of lcTAK1 and lcTABs, higher than that after the overexpression of lcTAK1 or lcTABs alone. Co-immunoprecipitation proved the direct interaction of lcTAB1/lcTAB2/lcTAB3 and lcTAK1. These findings indicated the roles of lcTABs in immune response of Larimichthys crocea.

20.
Cancer Biomark ; 24(3): 325-334, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30883339

RESUMO

Abnormally expressed microRNAs (miRNAs) contribute widely to human cancer, including oral squamous cell carcinoma (OSCC), by regulating their downstream targets. MiR-223 has been proved to be up-regulated in both gastric cancer and ovarian cancer. However, the effect of miR-223 on OSCC is still unclear. Here, we showed that miR-223 was over-expressed in OSCC tissues using qRT-PCR. Next, we investigated the biological mechanism of miR-223 in OSCC. The results demonstrated that miR-223 facilitated the cell proliferation and migration of OSCC using MTT assay and Transwell assay. Furthermore, we stated that the FBXW7 expression was decreased in OSCC and re-expression of FBXW7 inhibited the proliferation and migration of OSCC. In addition, FBXW7 mimic inversed the promotion effect of miR-223 in regulating of OSCC cells. In short, miR-223 promoted OSCC cell proliferation and migration by downregulating FBXW7, which provided a novel therapeutic strategy for OSCC.


Assuntos
Carcinoma de Células Escamosas/genética , Proteína 7 com Repetições F-Box-WD/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Neoplasias Bucais/genética , Interferência de RNA , Regiões 3' não Traduzidas , Adulto , Idoso , Apoptose/genética , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células , Progressão da Doença , Feminino , Genes Reporter , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/patologia , Estadiamento de Neoplasias , Carga Tumoral
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