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BACKGROUND: Ultrasound is a vital tool for the diagnosis and management of colorectal cancer (CRC). Contrast-enhanced ultrasound (CEUS) is a non-invasive, safe, and cost-effective method for evaluating tumour blood vessels, that play a crucial role in tumour growth and progression. AIM: To explore CEUS's role in the quantitative evaluation of CRC blood vessels and their correlation with angiogenesis markers and prognosis. METHODS: This study prospectively enrolled 100 patients with CRC confirmed by histopathology. All patients received preoperative CEUS examinations. Quantitative parameters, such as peak intensity (PI), time to peak (TTP), and area under the curve (AUC), were derived from time-intensity curve (TIC) analysis. Tumour tissue samples were obtained during surgery and examined immunohistochemically to assess the expression of angiogenesis markers, including vascular endothelial growth factor (VEGF) and microvessel density (MVD). The correlation between CEUS parameters, angiogenesis markers, and clinicopathological features was evaluated using appropriate statistical tests. RESULTS: Quantitative CEUS parameters (PI, TTP, and AUC) showed significant correlations with VEGF expression (P < 0.001) and MVD (P < 0.001), indicating a strong link between tumour blood vessels and angiogenesis. Increased PI, reduced TTP, and expanded AUC values were significantly related to higher tumour stage (P < 0.001), lymph node metastasis (P < 0.001), and distant metastasis (P < 0.001). Furthermore, these parameters were recognized as independent predictors of overall survival and disease-free survival in multivariate analysis (P < 0.001). CONCLUSION: CEUS has a high potential in guiding treatment planning and predicting patient outcomes. However, more comprehensive, multicentre studies are required to validate the clinical utility of CEUS in CRC management.
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BACKGROUND: Metazoan guts are in permanent contact with microbial communities. However, the host mechanisms that have developed to manage the dynamic changes of these microorganisms and maintain homeostasis remain largely unknown. RESULTS: Serotonin (5-hydroxytryptamine [5-HT]) was found to modulate gut microbiome homeostasis via regulation of a dual oxidase (Duox) gene expression in both Bactrocera dorsalis and Aedes aegypti. The knockdown of the peripheral 5-HT biosynthetic gene phenylalanine hydroxylase (TPH) increased the expression of Duox and the activity of reactive oxygen species, leading to a decrease in the gut microbiome load. Moreover, the TPH knockdown reduced the relative abundance of the bacterial genera Serratia and Providencia, including the opportunistic pathogens, S. marcescens and P. alcalifaciens in B. dorsalis. Treatment with 5-hydroxytryptophan, a precursor of 5-HT synthesis, fully rescued the TPH knockdown-induced phenotype. CONCLUSIONS: The findings reveal the important contribution of 5-HT in regulating gut homeostasis, providing new insights into gut-microbe interactions in metazoans.
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Microbioma Gastrointestinal , Animais , Microbioma Gastrointestinal/fisiologia , Homeostase , Insetos , Serotonina , SerratiaRESUMO
OBJECTIVE: To analyze the incidence and associated factors of hypothyroidism after radioiodine treatment for hyperthyroidism during a 13-year follow-up period. SUBJECTS AND METHODS: This was a retrospective study of consecutive patients with hyperthyroidism who were treated using a single dose of radioactive iodine (RAI) with a calculated dose regimen from 07/2005 to 12/2012. Univariate and multivariate Cox regression models were used to examine the factors that are associated with the occurrence of hypothyroidism after RAI therapy. Kaplan-Meier analysis was used for confirming associations between these models. RESULTS: A total of 182 patients were included during a 7.5-year median follow-up (range: 6-13 years). They were 36.4±11.1 years. The mean radioactive iodine dosage was 308.2±104.3 (range: 129.5-740.0) MBq. The rates of euthyroidism, early hypothyroidism, improvement, and ineffective treatment at 6 months were 48.4%, 37.9%, 8.8%, and 4.9%, respectively. The cumulative incidence of hypothyroidism in all patients with hyperthyroidism was 45.6% at 1 year, 48.9% at 5 years, and 52.3% at 10 years. Thyroid weight >46g (HR=0.643, 95%CI: 0.422-0.981, P=0.040) and a course of disease of 0.5-3 years (HR=0.592, 95%CI: 0.358-0.981, P=0.042) were identified as independent factors associated with an increased risk of hypothyroidism after radioactive iodine therapy. CONCLUSION: Radioactive iodine treatment with a calculated dose has a high cure rate for hyperthyroidism and has a low annual increase of hypothyroidism. Hypothyroidism after radioactive iodine treatment is more likely to occur in patients with small thyroid and a short disease course.
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Hipertireoidismo , Hipotireoidismo , Neoplasias da Glândula Tireoide , Seguimentos , Humanos , Hipertireoidismo/radioterapia , Hipotireoidismo/etiologia , Radioisótopos do Iodo/efeitos adversos , Estudos Retrospectivos , Neoplasias da Glândula Tireoide/complicações , Resultado do TratamentoRESUMO
OBJECTIVE: To investigate the value of 68 Ga-PSMA-11 positron emission tomography/computerized tomography (PET/CT) in evaluating lacrimal and salivary glands function. METHODS: Ten patients with pSS and 18 healthy volunteers were recruited in this study. All participants underwent 68 Ga-PSMA-11 PET/CT, and the patients with pSS performed salivary gland scintigraphy the next day. The maximum standardized uptake value (SUVmax), average of the standard uptake value (SUVavg), the average CT value (CTavg), and volume (V) in the region of interest (ROI) of each lacrimal and salivary gland were analyzed in68Ga-PSMA-11 PET/CT. The uptake ratio (UR) of the bilateral parotid gland and submandibular gland was calculated in salivary gland scintigraphy (SGS). Statistical analysis was processed by the SPSS software and the MedCalc software. A p-value of < 0.05 was considered as statistically significant. RESULTS: Almost all the parameters of pSS were significantly lower than those of the control group (p < 0.05). The left parotid gland (PG) UR was positive correlation with left PG SUVmax (r = 0.758, p = 0.011) and left PG SUVavg (r = 0.770, p = 0.009); the right PGUR was positive correlation with right PG SUVmax (r = 0.721, p = 0.019) and right PG SUVavg (r = 0.721, p = 0.019). The SUVmax and SUVavg of both sides of acrimal and salivary glands had area under the receiver operating curve values greater than 0.5. CONCLUSIONS: 68 Ga-PSMA-11 PET/CT can simultaneously enable the visualization of lacrimal glands and salivary glands and be used to evaluate the lacrimal and salivary glands function. Key Points ⢠We have firstly investigated the value of 68 Ga-PSMA-11 PET/CT in evaluating lacrimal and salivary glands function in patients with pSS 68 Ga-PSMA-11 PET/CT can simultaneously allow the visualization of lacrimal glands and salivary glands. ⢠The results of the present study imply that 68 Ga-PSMA-11 PET/CT can be used to evaluate the lacrimal and salivary glands function in patients with pSS meanwhile.
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Aparelho Lacrimal , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Humanos , Aparelho Lacrimal/diagnóstico por imagem , Glândula Parótida/diagnóstico por imagem , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Cintilografia , Glândulas Salivares/diagnóstico por imagemRESUMO
Muscarinic acetylcholine receptors (mAChRs) play important roles in the insect nervous system. These receptors are G protein-coupled receptors, which are potential targets for insecticide development. While the investigation of pharmacological properties of insect mAChRs is growing, the physiological roles of the receptor subtype remain largely indeterminate. Here, we identiï¬ed three mAChR genes in an important agricultural pest Bactrocera dorsalis. Phylogenetic analysis deï¬ned these genes as mAChR-A, -B, and -C. Transcripts of the three mAChRs are most prevalent in 1-d-old larvae and are more abundant in the brain than other body parts in adults. Functional assay of Bdor-mAChR-B transiently expressed in Chinese hamster ovary cells showed that it was activated by acetylcholine (EC50, 205.11 nM) and the mAChR agonist oxotremorine M (EC50, 2.39 µM) in a dose-dependent manner. Using the CRISPR/Cas9 technique, we successfully obtained a Bdor-mAChR-B knockout strain based on wild-type (WT) strain. When compared with WT, the hatching and eclosion rate of Bdor-mAChR-B mutants are significantly lower. Moreover, the crawl speed of Bdor-mAChR-B knockout larvae was lower than that of WT, while climbing performance was enhanced in the mutant adults. Adults with loss of function of Bdor-mAChR-B showed declined copulation rates and egg numbers (by mated females). Our results indicate that Bdor-mAChR-B plays a key role in the development, locomotion, and mating behavior of B. dorsalis.
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Acetilcolina/farmacologia , Proteínas de Insetos/genética , Agonistas Muscarínicos/farmacologia , Oxotremorina/análogos & derivados , Receptores Muscarínicos/genética , Tephritidae/genética , Animais , Sequência de Bases , Proteínas de Insetos/metabolismo , Masculino , Oxotremorina/farmacologia , Filogenia , Receptores Muscarínicos/metabolismo , Alinhamento de Sequência , Tephritidae/metabolismoRESUMO
BACKGROUND: This study aimed to assess the value of biphasic GA 68-labeled prostate-specific membrane antigen-11 (68Ga-PSMA-11) positron emission tomography/computed tomography (PET/CT) scan in the differential diagnosis and risk stratification of initial primary prostate cancer (PCa). METHODS: A total of 51 patients with PCa (8 low- and intermediate-risk PCa patients and 43 high-risk PCa patients) and 36 patients with benign prostate lesions, who underwent standard whole-body imaging and delayed pelvic imaging of 68Ga-PSMA-11 PET/CT, were enrolled in this prospective study. The PET parameters, such as maximum and mean standard uptake value (SUVmax and SUVmean), and maximum and mean standard retention index of PET images were calculated and compared in different prostate lesions. The diagnostic performances of the PET parameters were evaluated by receiver operating characteristic (ROC) curves. RESULTS: All the PET parameters of PCa participants were significantly higher than those of participants with benign prostate lesions (P<0.001). The SUVmean of delayed imaging had the best performance in the diagnosis of PCa with an area under the curve (AUC) of 0.918 (95% CI: 0.858 to 0.977), the sensitivity of 90.0%, and specificity of 83.3%. The SUVmax and SUVmean of high-risk PCa participants were significantly higher than those of low- and intermediate-risk PCa participants (P<0.005). The SUVmax of standard imaging had the best performance in predicting high-risk PCa with an AUC of 0.890 (95% CI: 0.799 to 0.980), a sensitivity of 76.7%, and a specificity of 100.0%. CONCLUSIONS: The biphasic 68Ga-PSMA-11 PET/CT scan had good performance in discriminating prostate cancer from benign prostate diseases. The SUVmean of the prostate lesion at delayed imaging of 68Ga-PSMA-11 PET/CT had the best value in the differential diagnosis of PCa, and the SUVmax at standard imaging was most valuable in predicting the risk stratification of PCa.
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PURPOSE: To compare the diagnostic accuracy of dual-phase 99mTc-MIBI single photon emission computed tomography/computed tomography (SPECT/CT) and 4D CT for the localization of hyperfunctioning parathyroid glands, a systematic review and meta-analysis was performed. Whether 4D CT combined to SPECT/CT [contrast-enhanced (CE)-SPECT/CT] had a better diagnostic performance than SPECT/CT alone in this scenario was also evaluated. MATERIAL AND METHODS: PubMed and Embase databases were searched for eligible studies. To reduce interstudy heterogeneity, only studies with clear head-to-head comparison were included. Publication bias was assessed by the Deeks funnel plot. The pooled sensitivity, specificity and the area under the curve (AUC) for 4D CT, SPECT/CT and CE-SPECT/CT were determined by random-effect analysis, respectively. RESULTS: Nine studies met the inclusion criteria, with a total of 911 participants. The sensitivity and specificity of 4D CT were 0.85 [95% confidence interval (CI), 0.69-0.94] and 0.93 (95% CI, 0.88-0.96), whereas the sensitivity and specificity for SPECT/CT were 0.68 (95% CI, 0.51-0.82; P = 0.048 compared with 4D CT) and 0.98 (95% CI, 0.95-0.99; P = 0.014 compared with 4D CT), respectively. CE-SPECT/CT is comparable to SPECT/CT in specificity and AUC, but it may improve the sensitivity (although there was a lack of statistical difference, 0.87 vs. 0.78; P = 0.125). CONCLUSION: Although 4D CT shows comparable AUC and borderline better sensitivity than SPECT/CT, its clinical application is confined by relatively low specificity and high radiation exposure. CE-SPECT/CT may improve the sensitivity without compromising the specificity and AUC of SPECT/CT.
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Tomografia Computadorizada Quadridimensional , Glândulas Paratireoides/diagnóstico por imagem , Glândulas Paratireoides/fisiopatologia , Tomografia Computadorizada com Tomografia Computadorizada de Emissão de Fóton Único , Tecnécio Tc 99m Sestamibi , Humanos , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Bactrocera dorsalis (Hendel) is the main fruit fly pest of tropical and subtropical countries. The application of insecticides to manage this pest has led to serious resistance problems; therefore, new ways to control B. dorsalis are required. Pathogenic bacteria are sources of biocontrol agents for pest management. RESULTS: We determined that a pathogenic bacterial strain, Serratia marcescens PS-1, isolated from a moribund striped flea beetle (Phyllotreta striolata), was lethal to B. dorsalis adults following ingestion. Histological analyses revealed that PS-1 damaged the intestinal epithelium, resulting in cell death within 24 h. We then generated a gut transcriptomic data set using RNA-Seq at two time points (6 and 24 h) after PS-1 infection. We found that genes encoding the peritrophic matrix constituent were down-regulated, whereas genes involved in lipid and glycan metabolism, and renewal of the gut epithelium, along with genes encoding digestive enzymes and stress response factors, were up-regulated. In addition, 14 cecropin genes were identified and cloned from B. dorsalis. To our knowledge, the number of cecropins identified in the present study is greater than that reported in the insects of earlier studies. Moreover, some of the cecropins identified were significantly down-regulated after PS-1 treatment. CONCLUSION: Our findings provide new insights into the insect gut response to pathogenic bacterial invasion and may aid the development of new strategies for the biological control of B. dorsalis. © 2019 Society of Chemical Industry.
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Tephritidae , Animais , Bactérias , Inseticidas , TranscriptomaRESUMO
Three new coordination polymers, [Cu2(CN)(L)(OCH3)]·CH3OH·3H2O (1), [Cu3(Br)3(L)] (2) and [Cu(I)(L)]·1.5H2O (3), have been solvothermally prepared by reacting L (5,11,17,23-tetra-tert-butyl-25,26,27,28-tetra[(3-pyridylmethyl)oxy]-2,8,14,20-tetrathiacalix[4]arene) with Cu(i) halide. 1 and 2 exhibit layers. 3 displays a chain, a supramolecular layer constructed by hydrogen bonds. The performance of 1-3 was examined as heterogeneous catalysts for azide-alkyne cycloaddition reactions. Most strikingly, 1 and 2 show predominant efficiency with high regioselectivity and excellent recyclability. Remarkably, solids 1-3 all have luminescence characteristics under irradiation with a UV lamp.
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Background & objectives: The nature of adaptable change of B-cell lymphoma-2 (BCL-2) and/or Bcl2-associated X protein (BAX) gene expression in the human peripheral blood mononuclear cells (PBMCs) irradiated by radioiodine in thyroid diseases therapy is not fully understood. In this study, the alternation of apoptotic gene expression was evaluated while the PBMCs collected from healthy volunteers were irradiated by the radioiodine-131 (131I). Methods: Fasting blood samples were obtained from healthy volunteers. PBMCs from group 0 to 6 were incubated and exposed to different doses of 131I in cell suspension for 6, 12, 24 and 48 h. The apoptosis rates and expression of BCL-2 and BAX genes of PBMCs were examined. Results: The apoptosis rate in the human PBMCs was gradually enhanced after six hour irradiation. The values of BCL-2 and BAX gene expression in groups 1-6 were higher than in group 0 within 6 h of irradiation, and then, these were decreased gradually from 6 to 12 h. BCL: -2 gene expression increased in groups 1-3 after 12 h irradiation, but there was no difference in groups 4-6. The ratio of BCL-2/BAX gene expression among groups 4-6 gradually decreased during the period from 6 to 12 h, and it was significantly lower than in the group 0 at 12, 24 and 48 h. Interpretation & conclusions: The expression of BCL-2 and BAX genes was initially upregulated following irradiation. Later, the balance of BCL-2/BAX genes expression was adjusted, and then, PBMCs underwent apoptosis at higher doses of radiation.
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Apoptose/efeitos da radiação , Leucócitos Mononucleares/efeitos da radiação , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteína X Associada a bcl-2/genética , Apoptose/genética , Regulação da Expressão Gênica/efeitos da radiação , Humanos , Radioisótopos do Iodo/efeitos adversos , Leucócitos Mononucleares/metabolismo , Radiação , Doses de RadiaçãoAssuntos
Adenoma/diagnóstico , Coristoma , Neoplasias Hepáticas/diagnóstico , Tumores Neuroendócrinos/diagnóstico , Pancreatopatias/patologia , Neoplasias Pancreáticas/diagnóstico , Baço , Adenoma/patologia , Adulto , Diagnóstico Diferencial , Feminino , Humanos , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/secundário , Tumores Neuroendócrinos/patologia , Pancreatectomia , Neoplasias Pancreáticas/patologiaRESUMO
To improve the specific activity and catalytic efficiency of l-LcLDH1, an NADH-dependent allosteric l-lactate dehydrogenase from L. casei, towards phenylpyruvic acid (PPA), its directed modification was conducted based on the semi-rational design. The three variant genes, Lcldh1Q88R, Lcldh1I229A and Lcldh1T235G, were constructed by whole-plasmid PCR as designed theoretically, and expressed in E. coli BL21(DE3), respectively. The purified mutant, l-LcLDH1Q88R or l-LcLDH1I229A, displayed the specific activity of 451.5 or 512.4 U/mg towards PPA, by which the asymmetric reduction of PPA afforded l-phenyllactic acid (PLA) with an enantiomeric excess (eep) more than 99%. Their catalytic efficiencies (kcat/Km) without d-fructose-1,6-diphosphate (d-FDP) were 4.8- and 5.2-fold that of l-LcLDH1. Additionally, the kcat/Km values of l-LcLDH1Q88R and l-LcLDH1I229A with d-FDP were 168.4- and 8.5-fold higher than those of the same enzymes without d-FDP, respectively. The analysis of catalytic mechanisms by molecular docking (MD) simulation indicated that substituting I229 in l-LcLDH1 with Ala enlarges the space of substrate-binding pocket, and that the replacement of Q88 with Arg makes the inlet of pocket larger than that of l-LcLDH1.
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Proteínas de Bactérias , L-Lactato Desidrogenase , Lacticaseibacillus casei/enzimologia , Ácidos Fenilpirúvicos/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Catálise , Escherichia coli/genética , Escherichia coli/metabolismo , L-Lactato Desidrogenase/química , L-Lactato Desidrogenase/genética , L-Lactato Desidrogenase/metabolismo , Simulação de Acoplamento Molecular , Mutagênese Sítio-Dirigida , Mutação , Ácidos Fenilpirúvicos/química , Engenharia de ProteínasRESUMO
Tumor metastasis is the leading cause of death in patients with advanced gastric cancer (GC). Limited therapeutic regimens are available for this condition, which is associated with a poor prognosis, and the mechanisms underlying tumor metastasis remain unclear. In the present study, increased histone methyltransferase G9A expression in GC tissues correlated with advanced stage and shorter overall survival, and in vitro and in vivo experiments revealed that G9A promoted tumor invasion and metastasis. Moreover, we observed that Reg IV induced G9A via the p-ERK/p-SP1 pathway. SP1 directly binds the G9A promoter and enhances G9A expression, and upregulated G9A then forms a transcriptional activator complex with P300 and GR, thereby promoting ITGB3 expression induced by dexamethasone (DEX) and contributing to GC metastasis. However, the G9A-mediated increase in ITGB3 expression was not dependent on the SET domain and methyltransferase activity of G9A. This study demonstrates that G9A is an independent prognostic marker and promotes metastasis in GC, thus suggesting that it may be a tumor biomarker and potential therapeutic target in GC.
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Biomarcadores Tumorais/metabolismo , Movimento Celular , Antígenos de Histocompatibilidade/metabolismo , Histona-Lisina N-Metiltransferase/metabolismo , Integrina beta3/metabolismo , Neoplasias Peritoneais/enzimologia , Neoplasias Gástricas/enzimologia , Animais , Sítios de Ligação , Biomarcadores Tumorais/genética , Linhagem Celular Tumoral , Proteína p300 Associada a E1A/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Feminino , Regulação Enzimológica da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Antígenos de Histocompatibilidade/genética , Histona-Lisina N-Metiltransferase/genética , Humanos , Integrina beta3/genética , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Invasividade Neoplásica , Domínios PR-SET , Proteínas Associadas a Pancreatite/metabolismo , Neoplasias Peritoneais/genética , Neoplasias Peritoneais/mortalidade , Neoplasias Peritoneais/secundário , Fosforilação , Regiões Promotoras Genéticas , Receptores de Glucocorticoides/metabolismo , Transdução de Sinais , Fator de Transcrição Sp1/metabolismo , Neoplasias Gástricas/genética , Neoplasias Gástricas/mortalidade , Neoplasias Gástricas/patologia , Regulação para CimaRESUMO
BACKGROUND: The ALEX is a novel member of the armadillo family and ALEX1 was reported to be reduced or even lost in multiple solid tumors. However, its expression profile and oncogenic role in gastric cancer (GC) remains largely unknown. METHODS: ALEX1 expression was detected in 161 GC samples by immunohistochemistry staining. NCI-N87 cells transfected by ALEX1 lentivirus vectors and MKN28 cells transfected by ALEX1 shRNA were used for biological function investigation. Western blot was applied to explore the molecular mechanism and pull-down assays were applied to measure the activity of Rho GTPases. In vivo tumorigenicity, peritoneal and lung metastasis experiments were performed by tumor cell engraftment into nude mice. Bisulfite genomic sequencing and methylation-specific PCR were applied to check the methylation status of the ALEX1 gene. RESULTS: The expression rate of ALEX1 was significantly reduced in gastric tumor samples compared to non-tumor samples (43.5 vs. 90.2%), and its expression was closely related to the tumor differentiation, TNM staging, and lymph nodes metastasis. ALEX1 overexpression in NCI-N87 cells significantly inhibited cell proliferation, migration, and invasion in vitro, and disrupted the structure of the cytoskeleton. ALEX1 overexpression attenuated xenografts growth, peritoneal, and lung metastasis in nude mice. Mechanistically, the overexpression of ALEX1 inhibits thrombin-induced metastasis and Rho GTPases activation. Bisulfite genomic sequencing and methylation-specific PCR revealed that the promoter of ALEX1 is highly methylated in GC cells and tissues. CONCLUSIONS: ALEX1 expression is reduced in GC and is involved in diverse cellular functions. ALEX1 inhibits metastasis through the PAR-1/Rho GTPase signaling pathway.
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Proteínas do Domínio Armadillo/genética , Proteínas Oncogênicas/genética , Receptor PAR-1/metabolismo , Neoplasias Gástricas/patologia , Proteínas rho de Ligação ao GTP/metabolismo , Animais , Linhagem Celular Tumoral , Proliferação de Células/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Metástase Linfática , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Metástase Neoplásica , Regiões Promotoras Genéticas , Transdução de Sinais , Neoplasias Gástricas/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Increasing evidence demonstrates that long noncoding RNAs (lncRNAs) regulate gene and protein expression by exerting an influence on transcriptional and post-transcriptional processes. Here, we report that the lncRNA UCA1 increases the metastatic ability of gastric cancer (GC) cells by regulating GRK2 protein stability by promoting Cbl-c-mediated GRK2 ubiquitination and degradation. This process then activates the ERK-MMP9 signalling pathway. Furthermore, we demonstrate that GRK2 is downregulated in GC cells and that silencing of GRK2 might cause similar phenotypic changes and signalling pathway activation as those induced by elevated UCA1 in GC cells. Our results suggest that UCA1 might function as a mediator of protein ubiquitination and may be a promising molecular target for GC therapy.
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Movimento Celular , Quinase 2 de Receptor Acoplado a Proteína G/metabolismo , Regulação Neoplásica da Expressão Gênica , Neoplasias Pulmonares/secundário , RNA Longo não Codificante/genética , Neoplasias Gástricas/patologia , Animais , Apoptose , Proliferação de Células , Quinase 2 de Receptor Acoplado a Proteína G/genética , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Masculino , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/genética , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Proteólise , Transdução de Sinais , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Long noncoding RNA UCA1 has emerged as a novel regulator in cancer initiation and progression of various cancers. However, function and underlying mechanism of UCA1 in the progression of gastric cancer (GC) remain unclear. In the present study, we report that UCA1 expressed highly in GC tissues and GC cells, which was partly induced by SP1. UCA1 promoted GC cell proliferation and G1/S transition in vitro and in vivo. Moreover, UCA1 exerted its function through interacting with EZH2, promoting direct interaction with cyclin D1 promoter to activate the translation of cyclin D1. Furthermore, AKT/GSK-3B/cyclin D1 axis was activated to upregulate cyclin D1 due to overexpression of UCA1. In addition, EZH2 and phosphorylated AKT induced by UCA1 could impact each other to form a positive feedback to promote cyclin D1 expression. This study demonstrated that UCA1 as a critical regulator involved in GC proliferation and cell cycle progression by promoting cyclin D1 expression, which indicates that it may be clinically a potential therapeutic target in GC.
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Ciclina D1/genética , Proteína Potenciadora do Homólogo 2 de Zeste/genética , Regulação Neoplásica da Expressão Gênica , Proteínas Proto-Oncogênicas c-akt/genética , RNA Longo não Codificante/genética , Fator de Transcrição Sp1/genética , Neoplasias Gástricas/genética , Idoso , Animais , Linhagem Celular Tumoral , Proliferação de Células , Ciclina D1/metabolismo , Progressão da Doença , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Retroalimentação Fisiológica , Feminino , Glicogênio Sintase Quinase 3 beta/genética , Glicogênio Sintase Quinase 3 beta/metabolismo , Humanos , Masculino , Camundongos , Camundongos Nus , Pessoa de Meia-Idade , Transplante de Neoplasias , Prognóstico , Regiões Promotoras Genéticas , Biossíntese de Proteínas , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Longo não Codificante/metabolismo , Transdução de Sinais , Fator de Transcrição Sp1/metabolismo , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/mortalidade , Neoplasias Gástricas/patologiaRESUMO
To improve the temperature characteristics and catalytic efficiency of a glycoside hydrolase family (GHF) 11 xylanase from Aspergillus oryzae (AoXyn11A), its variants were predicted based on in silico design. Firstly, Gly21 with the maximum B-factor value, which was confirmed by molecular dynamics (MD) simulation on the three-dimensional structure of AoXyn11A, was subjected to site-saturation mutagenesis. Thus, one variant with the highest thermostability, AoXyn11AG21I, was selected from the mutagenesis library, E. coli/Aoxyn11A G21X (X: any one of 20 amino acids). Secondly, based on the primary structure multiple alignment of AoXyn11A with seven thermophilic GHF11 xylanases, AoXyn11AY13F or AoXyn11AG21I-Y13F, was designed by replacing Tyr13 in AoXyn11A or AoXyn11AG21I with Phe. Finally, three variant-encoding genes, Aoxyn11A G21I, Aoxyn11A Y13F and Aoxyn11A G21I-Y13F, were constructed by two-stage whole-plasmid PCR method, and expressed in Pichia pastoris GS115, respectively. The temperature optimum (T opt) of recombinant (re) AoXyn11AG21I-Y13F was 60 °C, being 5 °C higher than that of reAoXyn11AG21I or reAoXyn11AY13F, and 10 °C higher than that of reAoXyn11A. The thermal inactivation half-life (t 1/2) of reAoXyn11AG21I-Y13F at 50 °C was 240 min, being 40-, 3.4- and 2.5-fold longer than those of reAoXyn11A, reAoXyn11AG21I and reAoXyn11AY13F. The melting temperature (T m) values of reAoXyn11A, reAoXyn11AG21I, reAoXyn11AY13F and reAoXyn11AG21I-Y13F were 52.3, 56.5, 58.6 and 61.3 °C, respectively. These findings indicated that the iterative mutagenesis of both Gly21Ile and Tyr13Phe improved the temperature characteristics of AoXyn11A in a synergistic mode. Besides those, the catalytic efficiency (k cat/K m) of reAoXyn11AG21I-Y13F was 473.1 mL mg-1 s-1, which was 1.65-fold higher than that of reAoXyn11A.
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BACKGROUND: Gastric cancer (GC) is one of the most malignant tumors and the second leading cause of cancer-related deaths in the world. Luteolin, a flavonoid present in many fruits and green plants, suppresses cancer progression. The effects of luteolin on GC cells and their underlying mechanisms remain unclear. METHODS: Effects of luteolin on cell proliferation, migration, invasion, and apoptosis were examined in vitro and in vivo by cell counting kit-8 (CCK-8), transwell assays, and flow cytometry, respectively. Real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blots were performed to evaluate Notch1 signaling and activation of epithelial-mesenchymal transition (EMT) in GC cells treated with or without luteolin. Immunohistochemistry was performed to examine proliferation and Notch1 expression in xenograft tumors. RESULTS: Luteolin significantly inhibited cell proliferation, invasion, and migration in a dose-dependent and time-dependent manner and promoted cell apoptosis. Luteolin reversed EMT by shrinking the cytoskeleton and by inducing the expression of epithelial biomarker E-cadherin and downregulating the mesenchymal biomarkers N-cadherin, vimentin and Snail. Furthermore, Notch1 signaling was inhibited by luteolin, and downregulation of Notch1 had similar effects as luteolin treatment on cell proliferation, migration, and apoptosis. In addition, luteolin suppressed tumor growth in vivo. A higher expression of Notch1 correlated with a poor overall survival and a poor time to first progression. Furthermore, co-immunoprecipitation analysis revealed that activated Notch1 and ß-catenin formed a complex and regulated cell proliferation, migration, and invasion. CONCLUSIONS: In this study, GC progression was inhibited by luteolin through suppressing Notch1 signaling and reversing EMT, suggesting that luteolin may serve as an effective anti-tumor drug in GC treatment.
Assuntos
Progressão da Doença , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Luteolina/uso terapêutico , Receptores Notch/metabolismo , Transdução de Sinais/efeitos dos fármacos , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/patologia , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/metabolismo , Luteolina/química , Luteolina/farmacologia , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , Modelos Biológicos , Invasividade Neoplásica , Prognóstico , Ensaio Tumoral de Célula-TroncoRESUMO
Immunoglobulin G4-related lung disease (IgG4-RLD) is a disease in which abundant activated IgG4-positive plasma cells and lymphocytes infiltrate lung tissues with high 18F-fluorodeoxyglucose uptake. Although various forms of radiologic features of IgG4-RLD have been reported, cavitating mass is a rare imaging feature and should be differentiated from cancer. Therefore, in this article, we report two cases both with unprovoked cough, bloody sputum and presenting quite similar cavitating lesions with high 18F-fluorodeoxyglucose uptake on positron emission tomography/ computed tomography, one of which diagnosed as IgG4-RLD and the other as lung cancer based on biopsy eventually. The awareness of the imaging features of IgG4-RLD and lung cancer described in the present study may help physicians to distinguish one from the other. IgG4-RLD should be considered in the differential diagnosis of cavitary lung lesions.
RESUMO
Biglycan (BGN) is an important component of the extracellular matrix (ECM) that is implicated in a variety of human cancers. In our previous study, we reported that BGN was overexpressed in gastric cancer (GC) tissues and promoted cancer metastasis. Moreover, the tubular formation capacity in HUVECs was promoted by stimulation with culture media from BGN-overexpressing GC cells, but the exact underlying mechanism is still unknown. The purpose of this study was to determine the role and molecular mechanism of BGN in VEGF expression in endothelial cells. We found that BGN stimulation of endothelial cells increased the interaction between NF-kB and the HIF-1α promoter, leading to enhanced promoter activity and increased HIF-1α mRNA levels, as well as augmented HIF-1 activity that resulted in VEGF expression. All of this was dependent on the interaction of BGN with its receptors, TLR2 and TLR4. Moreover, we found that BGN enhanced endothelial cell migration and proliferation, as well as tube formation, in a TLR signaling pathway-dependent manner. In addition, endothelial cell-derived VEGF in turn was found to act on GC cells and promotes their migration. The combined findings of our current and previous studies suggest that BGN secreted from GC cells into the tumor stroma promotes GC development, as well as its progression, potentially through the chronic activation of tumor angiogenesis.