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BACKGROUND: Resistance to chemotherapeutic drugs limits the control of gastric cancer (GC) development. The study intended to probe into the mechanism of aquaporin 3 (AQP3) on the chemoresistance of GC. METHODS: Cisplatin (CDDP)-resistant cells were constructed. Parental AGS and HGC-27 cells and their respective CDDP-resistant cells were transfected with AQP3 overexpression plasmid, AQP3 short hairpin RNA (sh-AQP3) and sh-Kruppel-like factor 5 (shKLF5). The expressions of AQP3 and factors related to autophagy (LC3 I, LC3 II, Atg5, Beclin-1, p62)/epithelial-mesenchymal transition (EMT; E-cadherin and snail) were assessed by Western blot and qRT-PCR. Cell counting kit-8 assay was adopted to test cell viability and half maximal inhibitory concentration (IC 50) was determined. Transwell assay was used for the examination of cell migration and invasion. The regulatory relationship of AQP3 and KLF5 was tested by chromatin immunoprecipitation (ChIP) and dual luciferase reporter assays. RESULTS: AQP3 was highly-expressed in GC cells and its level was even higher in CDDP-resistant GC cells. AQP3 silencing inhibited viability, autophagy and EMT in CDDP-resistant GC cells, while AQP3 overexpression had the opposite effect. KLF5 positively modulated AQP3 in GC cells resistant to CDDP. KLF5 knockdown reversed AQP3-induced autophagy, viability, migration, invasion and EMT in CDDP-resistant GC cells. CONCLUSION: KLF5-modulated AQP3 activated autophagy to facilitate the resistance of GC to CDDP.
Assuntos
Cisplatino , Neoplasias Gástricas , Humanos , Cisplatino/farmacologia , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Aquaporina 3 , Fatores de Transcrição/metabolismo , Autofagia , Proliferação de Células , Linhagem Celular Tumoral , Fatores de Transcrição Kruppel-Like/genética , Fatores de Transcrição Kruppel-Like/metabolismo , Fatores de Transcrição Kruppel-Like/farmacologiaRESUMO
OBJECTIVE: To profile gut microbiome-associated metabolites in serum and investigate whether these metabolites could distinguish individuals with colorectal cancer (CRC) or adenoma from normal healthy individuals. DESIGN: Integrated analysis of untargeted serum metabolomics by liquid chromatography-mass spectrometry and metagenome sequencing of paired faecal samples was applied to identify gut microbiome-associated metabolites with significantly altered abundance in patients with CRC and adenoma. The ability of these metabolites to discriminate between CRC and colorectal adenoma was tested by targeted metabolomic analysis. A model based on gut microbiome-associated metabolites was established and evaluated in an independent validation cohort. RESULTS: In total, 885 serum metabolites were significantly altered in both CRC and adenoma, including eight gut microbiome-associated serum metabolites (GMSM panel) that were reproducibly detected by both targeted and untargeted metabolomics analysis and accurately discriminated CRC and adenoma from normal samples. A GMSM panel-based model to predict CRC and colorectal adenoma yielded an area under the curve (AUC) of 0.98 (95% CI 0.94 to 1.00) in the modelling cohort and an AUC of 0.92 (83.5% sensitivity, 84.9% specificity) in the validation cohort. The GMSM model was significantly superior to the clinical marker carcinoembryonic antigen among samples within the validation cohort (AUC 0.92 vs 0.72) and also showed promising diagnostic accuracy for adenomas (AUC=0.84) and early-stage CRC (AUC=0.93). CONCLUSION: Gut microbiome reprogramming in patients with CRC is associated with alterations of the serum metabolome, and GMSMs have potential applications for CRC and adenoma detection.
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Adenoma , Neoplasias Colorretais , Microbioma Gastrointestinal , Adenoma/diagnóstico , Biomarcadores Tumorais , Neoplasias Colorretais/genética , Microbioma Gastrointestinal/genética , Humanos , Metaboloma , MetagenomaRESUMO
BACKGROUND: The potential advantages of laparoscopic-assisted total gastrectomy (LATG) compared with open total gastrectomy (OTG) for Siewert Types II and III adenocarcinoma of the esophagogastric junction (AEJ) are not very clear. Thus, the aim of this study was to investigate the surgical outcomes and potential advantages of LATG for Siewert Types II and III AEJ. METHODS: The clinical data of 75 patients (32 for LATG and 43 for OTG) with Siewert II or III AEJ from August 2009 to February 2014 were analyzed retrospectively. Patients were followed up by telephone or out-patient examination till August 2015. RESULTS: Two groups of patients were successfully performed with no perioperative death. The mean operation time was 3.23 ± 0.35 hr in LATG group, longer than the OTG group 2.83 ± 0.51 hr. The mean intraoperative bleeding was 122.7 ± 50.6 ml, less than the OTG group 219.2 ± 85.2 ml. The analgesics use was 3.00 ± 0.67 times in the LATG group, less than the OTG group 3.43 ± 1.03 times. The gastrointestinal function recovery time was 2.69 ± 0.46 days in the LATG group, shorter than the OTG group 3.42 ± 0.86 days. The mean postoperative hospital stay was 12.94 + 2.76 days in the LATG group, less than the OTG group 14.57 + 2.35 days (p < 0.05). CONCLUSIONS: LATG and OTG had no significant difference for Siewert II and III AEJ in terms of radical resection and tumor recurrence, but LATG is worthy to be promoted with less bleeding, less postoperative pain, faster recovery of gastrointestinal function, and shorter hospital stay.
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Adenocarcinoma/cirurgia , Neoplasias Esofágicas/cirurgia , Junção Esofagogástrica/patologia , Junção Esofagogástrica/cirurgia , Gastrectomia/métodos , Laparoscopia/métodos , Adenocarcinoma/patologia , Antígeno Carcinoembrionário/sangue , Neoplasias Esofágicas/patologia , Feminino , Humanos , Excisão de Linfonodo , Linfonodos/patologia , Linfonodos/cirurgia , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/patologia , Estudos Retrospectivos , Neoplasias Gástricas/patologia , Neoplasias Gástricas/cirurgia , Resultado do TratamentoRESUMO
Endoplasmic reticulum resident protein 44 (ERp44) is a member of the protein disulfide isomerase family and functions in oxidative protein folding in the endoplasmic reticulum. A structurally flexible C-terminal tail (C-tail) of ERp44 plays critical roles in dynamically regulating ERp44's function in protein folding quality control. The structure-function dynamics of ERp44's C-tail may be studied further using fluorescence and other techniques, if methods are found to label the C-tail site-specifically with a fluorescent group or segmentally with other desired labels. Here we have developed such methods, employing split inteins capable of protein trans-splicing, and identifying atypical S1 split inteins able to function efficiently at a suitable split site in the ERp44 sequence. One method demonstrated segmental expression of ERp44 for segmental labeling of the C-tail, another method efficiently added a commercially available fluorescent group to the C-terminus of ERp44, and both methods may also be generally useful for studying other proteins.
Assuntos
Escherichia coli/genética , Inteínas , Proteínas de Membrana/genética , Chaperonas Moleculares/genética , Processamento de Proteína , Sequência de Aminoácidos , Fluorescência , Corantes Fluorescentes/análise , Corantes Fluorescentes/metabolismo , Humanos , Proteínas de Membrana/análise , Chaperonas Moleculares/análise , Dados de Sequência Molecular , Proteínas Recombinantes de Fusão/análise , Proteínas Recombinantes de Fusão/genética , Trans-SplicingRESUMO
Site-specific protein labeling are powerful means of protein research and engineering; however, new and improved labeling methods are greatly needed. Split inteins catalyze a protein trans-splicing reaction that can be used for enzymatic and nearly seamless protein labeling. Non-canonical S11 split intein has been used in an earlier method of protein C-terminal labeling; however, its relatively large (~150 aa) N-intein fused to the target protein often hindered protein expression, folding, and solubility. To solve this problem, here, we have designed and demonstrated a new method of protein C-terminal labeling, by first engineering a functional non-canonical S1 split intein that has an extremely small (12 aa) N-intein and a cysteine-free C-intein. An engineered Rma DnaB S1 split intein was modified to have a cysteine-free C-intein, while still retaining its robust trans-splicing function, which permitted the C-extein in a C-precursor to have a single cysteine for easy and specific linkage with desired labeling groups. The resulting new and generally useful method has two unique advantages: (1) The extremely small (12 aa) N-intein, which must be fused to the C terminus of the target protein, is less likely to hinder the protein expression, folding, and solubility; and (2) the single cysteine in the C-extein may be readily linked to a variety of labeling or modification groups using commercially available reagents.
Assuntos
Inteínas , Proteína C/genética , Motivos de Aminoácidos , Sequência de Aminoácidos , Cisteína/química , Cisteína/genética , Cisteína/metabolismo , Cinética , Dados de Sequência Molecular , Proteína C/química , Proteína C/metabolismo , Processamento de Proteína , Trans-SplicingRESUMO
Our study mainly reports the specific mechanisms of microRNA (miR)-874-3p on drug resistance in gastric cancer (GC). Clinical specimen was collected. The upstream long non-coding RNA (lncRNA) and the downstream gene of miR-874-3p were predicted using bioinformatic analysis with the results being ascertained with dual-luciferase reporter assay. The viability, apoptosis, migration and invasion of transfected GC cells with or without cisplatin (DDP) treatment were evaluated by Cell Counting Kit-8 (CCK-8), flow cytometric, Scratch, and Transwell assays. An animal xenograft model was constructed. Expressions of long intergenic non-coding RNA 00922 (LINC00922), miR-874-3p and potential target genes were quantified by quantitative real-time polymerase-chain reaction (qRT-PCR) and Western blot. MiR-874-3p, which was lower-expressed in drug-resistant GC tissues and cells, was upregulated to repress the viability, migration and invasion but enhance the apoptosis and sensitivity in GC cells with or without DDP resistance. Downregulation of miR-874-3p eliminated the effects of silenced LINC00922, a upstream lncRNA of miR-874-3p, on cell viability, apoptosis, migration and invasion, as well as the expressions of Glycerophosphodiester Phosphodiesterase Domain Containing 5 (GDPD5) and the downstream gene of miR-874-3p in DDP-resistant GC cells. GDPD5 silencing diminished the effects of miR-874-3p downregulation on GDPD5 expression, viability, migration and invasion of DDP-resistant GC cells. Additionally, LINC00922 silencing enhanced the inhibitory effect of DDP on tumor growth, whereas reversing the effects of DDP on LINC00922, miR-874-3p and GDPD5 expressions in tumors. MiR-874-3p, an miRNA, which is sponged by LINC00922 and targets GDPD5, inhibits the GC progression yet enhances the DDP sensitivity in GC.
Assuntos
MicroRNAs , RNA Longo não Codificante , Neoplasias Gástricas , Animais , Apoptose , Linhagem Celular Tumoral , Proliferação de Células/genética , Cisplatino/farmacologia , Resistencia a Medicamentos Antineoplásicos/genética , Humanos , MicroRNAs/metabolismo , Diester Fosfórico Hidrolases , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismoRESUMO
During the development of a pharmaceutical formulation, a powerful tool is needed to extract the key points from the complicated process parameters and material attributes. Artificial neural networks (ANNs), a promising and more flexible modeling technique, can address real intricate questions in a high parallelism and distributed pattern in the manner of biological neural networks. The data mined and analyzing based on ANNs have the ability to replace hundreds of trial and error experiments. ANNs have been used for data analysis by pharmaceutics researchers since the 1990s and it has now become a research method in pharmaceutical science. This review focuses on the latest application progress of ANNs in the prediction, characterization and optimization of pharmaceutical formulation to provide a reference for the further interdisciplinary study of pharmaceutics and ANNs.
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Chromatin immunoprecipitation (ChIP) studies were conducted in human hepatocytes treated with rifampicin in order to identify new pregnane-X receptor (PXR) target genes. Genes, both previously known to be involved and not known to be involved in drug disposition, with PXR response elements (PXREs) located upstream, within or downstream from their potentially associated genes, were identified. Validation experiments identified several new drug disposition genes with PXR binding sites. Of these, only CYP4F12 demonstrated increased binding in the presence of rifampicin. The role of PXR in the basal and inductive response of CYP4F12 was confirmed in hepatocytes in which PXR was silenced. We also assessed the association of PXR-coactivators and -corepressors with known and newly identified PXREs. Both PXR and the steroid receptor coactivator (SRC-1) were found to bind to PXREs in the absence of rifampicin, although binding was stronger after rifampicin treatment. We observed promoter-dependent patterns with respect to the binding of various coactivators and corepressors involved in the regulation of CYP4F12, CYP3A4, CYP2B6, UGT1A1 and P-glycoprotein. In conclusion, our findings indicate that PXR is involved in the regulation of CYP4F12 and that PXR along with SRC1 binds to a broad range of promoters but that many of these are not inducible by rifampicin.
Assuntos
Hepatócitos/metabolismo , Regiões Promotoras Genéticas , Receptores de Esteroides/metabolismo , Proteínas Repressoras/metabolismo , Transativadores/metabolismo , Adulto , Sítios de Ligação , Imunoprecipitação da Cromatina , Feminino , Hepatócitos/efeitos dos fármacos , Humanos , Pessoa de Meia-Idade , Receptor de Pregnano X , Interferência de RNA , Receptores de Esteroides/antagonistas & inibidores , Receptores de Esteroides/genética , Elementos de Resposta , Rifampina/farmacologia , Ativação TranscricionalRESUMO
Gastric cancer is one of the most common and deadly neoplasms with limited effective treatments. The emergence of the immunotherapy has brought great expectations for cancer patients. Sophoridine is extracted from the seeds of sophora alopecuroides and has various pharmacological actions including anti-tumor, anti-inflammatory, anti- arrhythmia and anti-virus. However, the effect of Sophoridine on gastric cancer microenvironment immunity and its underling mechanism remains poorly known. This study was aimed to investigate the effect of Sophoridine on the polarization status of gastric tumor-associated macrophages (TAMs) and its underlying mechanism. We isolated primary bone marrow-derived macrophages (BMDMs) and primary CD8+ T cells to perform coculture assay. Sophoridine educated TAMs polarize to M1-TAMs and suppressed M2-TAMs polarization through TLR4/IRF3 axis. Sophoridine-treated TAMs exhibited stronger pro-inflammatory function through upregulation the expression of INOS, IFN-ß and IL-12α, and downregulation the expression of Arg-1, CD206 and IL-10. Sophoridine -primed TAMs increased the proliferation and cytotoxic function of CD8+ T by upregulating the expression of Granzyme-B, TNF-α and Perforin, and downregulated the expression of CD8+ T cells function exhaustion markers PD-1, Tim-3 and Lag-3. Furthermore, Sophoridine inhibited the migration ability of macrophage by decrease the CCR2 expression. Thus, Sophoridine acted on macrophages and CD8+ T cells to reshape gastric cancer immune microenvironment. Our studies provided preclinical basis for clinical application of Sophoridine.
Assuntos
Alcaloides/farmacologia , Antineoplásicos/farmacologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Fator Regulador 3 de Interferon/metabolismo , Macrófagos/efeitos dos fármacos , Quinolizinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Neoplasias Gástricas/tratamento farmacológico , Receptor 4 Toll-Like/metabolismo , Animais , Linfócitos T CD8-Positivos/metabolismo , Linhagem Celular , Linhagem Celular Tumoral , Regulação para Baixo/efeitos dos fármacos , Humanos , Tolerância Imunológica/efeitos dos fármacos , Terapia de Imunossupressão/métodos , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Células RAW 264.7 , Neoplasias Gástricas/metabolismo , Microambiente Tumoral/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismo , Regulação para Cima/efeitos dos fármacos , MatrinasRESUMO
Circular RNAs (circRNAs) have recently emerged as novel and potentially promising therapeutic targets in a serious of cancers. However, the expression pattern and biological function of circRNAs in colon cancer remain largely elusive. This study firstly analyzed circRNA microarray of colon cancer and selected circ-0001313 as the study object. We aim to comprehensively investigate the expression pattern and biological function of circ-0001313 in the progression of colon cancer. Relative levels of circ-0001313 and miRNA-510-5p in colon cancer tissues and cell lines were determined with qRT-PCR. The binding relationship between miRNA-510-5p to circ-0001313 and AKT2 was predicted by bioinformatics analyses and further confirmed by dual-luciferase reporter gene assay. Regulatory effects of circ-0001313/miRNA-510-5p/AKT2 axis on colon cancer cells were evaluated by EdU assay and flow cytometry. Consistent with the microarray analysis, circ-0001313 was highly expressed in colon cancer tissues and cell lines. Knockdown of circ-0001313 attenuated proliferative ability, but induced apoptosis of colon cancer cells. Furthermore, we confirmed that circ-0001313 competitively bound to miRNA-510-5p, thus upregulating its target gene AKT2. Moreover, western blot analyses revealed that circ-0001313 also affects the expression of Bcl-2 family proteins and the activation of PI3K/Akt signaling pathway. In conclusion, our study revealed that circ-0001313 regulates the pathogenesis of colon cancer by sponging miRNA-510-5p to upregulate AKT2 expression.
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Uncovering pathways underlying drug-induced toxicity is a fundamental objective in the field of toxicogenomics. Developing mechanism-based toxicity biomarkers requires the identification of such novel pathways and the order of their sufficiency in causing a phenotypic response. Genome-wide RNA interference (RNAi) phenotypic screening has emerged as an effective tool in unveiling the genes essential for specific cellular functions and biological activities. However, eliciting the relative contribution of and sufficiency relationships among the genes identified remains challenging. In the rodent, the most widely used animal model in preclinical studies, it is unrealistic to exhaustively examine all potential interactions by RNAi screening. Application of existing computational approaches to infer regulatory networks with biological outcomes in the rodent is limited by the requirements for a large number of targeted permutations. Therefore, we developed a two-step relay method that requires only one targeted perturbation for genome-wide de novo pathway discovery. Using expression profiles in response to small interfering RNAs (siRNAs) against the gene for peroxisome proliferator-activated receptor alpha (Ppara), our method unveiled the potential causal sufficiency order network for liver hypertrophy in the rodent. The validity of the inferred 16 causal transcripts or 15 known genes for PPARalpha-induced liver hypertrophy is supported by their ability to predict non-PPARalpha-induced liver hypertrophy with 84% sensitivity and 76% specificity. Simulation shows that the probability of achieving such predictive accuracy without the inferred causal relationship is exceedingly small (p < 0.005). Five of the most sufficient causal genes have been previously disrupted in mouse models; the resulting phenotypic changes in the liver support the inferred causal roles in liver hypertrophy. Our results demonstrate the feasibility of defining pathways mediating drug-induced toxicity from siRNA-treated expression profiles. When combined with phenotypic evaluation, our approach should help to unleash the full potential of siRNAs in systematically unveiling the molecular mechanism of biological events.
Assuntos
Inativação Gênica , Fígado/metabolismo , Fígado/patologia , Modelos Biológicos , PPAR alfa/metabolismo , Proteoma/metabolismo , RNA Interferente Pequeno/genética , Animais , Simulação por Computador , Perfilação da Expressão Gênica/métodos , Hipertrofia/induzido quimicamente , Hipertrofia/metabolismo , Fígado/efeitos dos fármacos , Camundongos , PPAR alfa/genética , RNA Interferente Pequeno/administração & dosagem , Transdução de SinaisRESUMO
RNA interference (RNAi) has great potential as a tool for studying gene function in mammals. However, the specificity and magnitude of the in vivo response to RNAi remains to be fully characterized. A molecular and phenotypic comparison of a genetic knockout mouse and the corresponding knockdown version would help clarify the utility of the RNAi approach. Here, we used hydrodynamic delivery of small interfering RNA (siRNA) to knockdown peroxisome proliferator activated receptor alpha (Ppara), a gene that is central to the regulation of fatty acid metabolism. We found that Ppara knockdown in the liver results in a transcript profile and metabolic phenotype that is comparable to those of Ppara-/- mice. Combining the profiles from mice treated with the PPARalpha agonist fenofibrate, we confirmed the specificity of the RNAi response and identified candidate genes proximal to PPARalpha regulation. Ppara knockdown animals developed hypoglycemia and hypertriglyceridemia, phenotypes observed in Ppara-/- mice. In contrast to Ppara-/- mice, fasting was not required to uncover these phenotypes. Together, these data validate the utility of the RNAi approach and suggest that siRNA can be used as a complement to classical knockout technology in gene function studies.
Assuntos
PPAR alfa/genética , Interferência de RNA , Animais , Perfilação da Expressão Gênica , Injeções , Fígado/metabolismo , Camundongos , Camundongos Knockout , PPAR alfa/metabolismo , Fenótipo , RNA Interferente Pequeno/administração & dosagem , Transcrição GênicaRESUMO
In this paper, efficient phosphorescent white organic light-emitting diodes (WOLEDs) were fabricated based on ultrathin doping-free emissive layers and mixed bipolar interlayers. The energy transfer processes were proved via the research of WOLEDs with different interlayer thicknesses and transient photoluminescence lifetime. WOLEDs with optimized thickness of doping-free emissive layers show maximum current efficiency of 47.8 cd/A and 44.9 cd/A for three-colors and four-colors WOLEDs, respectively. The Commission Internationale de L'Eclairage coordinates shows a very slight variation of ( ± 0.02, ± 0.02) from 5793 cd/m2 to 11370 cd/m2 for three-colors WOLEDs and from 3038 cd/m2 to 13720 cd/m2 for four-colors WOLEDs, respectively. The stability of the spectra is attributed to the stable and sequential energy transfer among the various dyes. The color temperature of four-colors WOLEDs can be obtained from 2659 to 6636 by adjusting the thickness of ultrathin emissive layer.
RESUMO
In rodents, treatment with peroxisome proliferator-activated receptor alpha (PPARalpha) agonists results in peroxisome proliferation, hepatocellular hypertrophy, and hepatomegaly. Drugs in the fibrate class of PPARalpha agonists have also been reported to produce rare skeletal muscle toxicity. Although target-driven hepatic effects of PPARalpha treatment have been extensively studied, a characterization of the transcriptional effects of this nuclear receptor/transcription factor on skeletal muscle responses has not been reported. In this study we investigated the effects of PPARalpha agonists on skeletal muscle gene transcription in rats. Further, since statins have been reported to preferentially effect type II muscle fibers, we compared PPARalpha signaling effects between type I and type II muscles. By comparing the transcriptional responses of agonists that signal through different nuclear receptors and using a selection/deselection analytical strategy based on ANOVA, we identified a PPARalpha activation signature that is evident in type I (soleus), but not type II (quadriceps femoris), skeletal muscle fibers. The fiber-type-selective nature of this response is consistent with increased fatty acid uptake and beta-oxidation, which represent the major clinical benefits of the hypolipidemic compounds used in this study, but does not reveal any obvious off-target pathways that may drive adverse effects.
Assuntos
Fibras Musculares de Contração Lenta/efeitos dos fármacos , Músculo Esquelético/efeitos dos fármacos , PPAR alfa/agonistas , Animais , Bezafibrato/farmacologia , Ácidos Graxos/metabolismo , Feminino , Fenofibrato/farmacologia , Perfilação da Expressão Gênica , Glucose/metabolismo , Hipolipemiantes/farmacologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fibras Musculares de Contração Lenta/metabolismo , Músculo Esquelético/metabolismo , Pirimidinas/farmacologia , Ratos , Ratos Endogâmicos , Rosiglitazona , Tiazolidinedionas/farmacologia , Tretinoína/farmacologiaRESUMO
Toxicity is a major cause of failure in drug development. A toxicogenomic approach may provide a powerful tool for better assessing the potential toxicity of drug candidates. Several approaches have been reported for predicting hepatotoxicity based on reference compounds with well-studied toxicity mechanisms. We developed a new approach for assessing compound-induced liver injury without prior knowledge of a compound's mechanism of toxicity. Using samples from rodents treated with 49 known liver toxins and 10 compounds without known liver toxicity, we derived a hepatotoxicity score as a single quantitative measurement for assessing the degree of induced liver damage. Combining the sensitivity of the hepatotoxicity score and the power of a machine learning algorithm, we then built a model to predict compound-induced liver injury based on 212 expression profiles. As estimated in an independent data set of 54 expression profiles, the built model predicted compound-induced liver damage with 90.9% sensitivity and 88.4% specificity. Our findings illustrate the feasibility of ab initio estimation of liver toxicity based on transcriptional profiles.
Assuntos
Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Perfilação da Expressão Gênica , Fígado/efeitos dos fármacos , Toxicogenética/métodos , Transcrição Gênica , Alanina Transaminase/sangue , Algoritmos , Animais , Inteligência Artificial , Aspartato Aminotransferases/sangue , Bilirrubina/sangue , Química Clínica/métodos , Colesterol/sangue , Análise por Conglomerados , Relação Dose-Resposta a Droga , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/induzido quimicamente , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/classificação , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/metabolismo , Estudos de Viabilidade , Fígado/enzimologia , Fígado/patologia , Modelos Biológicos , Preparações Farmacêuticas/classificação , Ratos , Ratos Sprague-Dawley , Sensibilidade e EspecificidadeRESUMO
To understand the heterogeneity of prostate cancer (PCa) and identify novel underlying drivers, we constructed integrative molecular Bayesian networks (IMBNs) for PCa by integrating gene expression and copy number alteration data from published datasets. After demonstrating such IMBNs with superior network accuracy, we identified multiple sub-networks within IMBNs related to biochemical recurrence (BCR) of PCa and inferred the corresponding key drivers. The key drivers regulated a set of common effectors including genes preferentially expressed in neuronal cells. NLGN4Y-a protein involved in synaptic adhesion in neurons-was ranked as the top gene closely linked to key drivers of myogenesis subnetworks. Lower expression of NLGN4Y was associated with higher grade PCa and an increased risk of BCR. We show that restoration of the protein expression of NLGN4Y in PC-3 cells leads to decreased cell proliferation, migration and inflammatory cytokine expression. Our results suggest that NLGN4Y is an important negative regulator in prostate cancer progression. More importantly, it highlights the value of IMBNs in generating biologically and clinically relevant hypotheses about prostate cancer that can be validated by independent studies.
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Teorema de Bayes , Moléculas de Adesão Celular Neuronais/genética , Variações do Número de Cópias de DNA , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes/genética , Neoplasias da Próstata/genética , Moléculas de Adesão Celular Neuronais/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Bases de Dados Genéticas , Progressão da Doença , Perfilação da Expressão Gênica , Humanos , Masculino , Neurônios/metabolismo , Neoplasias da Próstata/patologia , Neoplasias da Próstata/terapiaRESUMO
Traditionally, the primary function of oligodendrocytes (OLGs) in the CNS has been considered to be myelination. Here, we investigated whether OLGs may play a trophic role, particularly during development. Neurotrophin expression was assessed in postnatal day 7 basal forebrain (BF) OLGs, using in situ hybridization and detection of myelin basic protein. Nerve growth factor, brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) mRNAs were revealed in OLGs in vivo and in culture. To determine whether OLGs support nearby neurons, we examined the influence of OLGs on BF cholinergic neurons. Neuronal function was enhanced by cocultured OLGs and OLG conditioned medium. Moreover, trophic effects of OLG conditioned medium were partially blocked by K252a, a trk tyrosine kinase inhibitor, and by neutralizing anti-BDNF or anti-NT-3 antisera, indicating that neurotrophins may mediate these effects, perhaps in concert with other signals. Our studies support a novel role for OLGs in providing local trophic support for neurons in the CNS.
Assuntos
Oligodendroglia/fisiologia , Prosencéfalo/crescimento & desenvolvimento , Prosencéfalo/fisiologia , Animais , Astrócitos/citologia , Astrócitos/fisiologia , Fator Neurotrófico Derivado do Encéfalo/antagonistas & inibidores , Fator Neurotrófico Derivado do Encéfalo/biossíntese , Fator Neurotrófico Derivado do Encéfalo/genética , Carbazóis/farmacologia , Divisão Celular/efeitos dos fármacos , Divisão Celular/fisiologia , Linhagem da Célula , Células Cultivadas , Fibras Colinérgicas/efeitos dos fármacos , Fibras Colinérgicas/fisiologia , Técnicas de Cocultura , Meios de Cultivo Condicionados/farmacologia , Inibidores Enzimáticos/farmacologia , Soros Imunes/farmacologia , Alcaloides Indólicos , Microglia/citologia , Microglia/fisiologia , Proteína Básica da Mielina/biossíntese , Fator de Crescimento Neural/antagonistas & inibidores , Fator de Crescimento Neural/biossíntese , Fator de Crescimento Neural/genética , Neurônios/citologia , Neurônios/efeitos dos fármacos , Neurônios/fisiologia , Neurotrofina 3/antagonistas & inibidores , Neurotrofina 3/biossíntese , Neurotrofina 3/genética , Oligodendroglia/citologia , Oligodendroglia/metabolismo , Prosencéfalo/citologia , RNA Mensageiro/biossíntese , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND: It is not clear yet about the secular changes of morbidity and mortality trend of lung cancer in residents of Nangang District of Harbin in China. The aim of this study is to estimate the trend of lung cancer morbidity and mortality in residents of Nangang District from 1992 to 2001 and to predict their levels in the future 5 years. METHODS: Data were collected from the annual statistic reports on cancer death cause from Nangang District in Harbin. The classification of death cause was made according to the ICD-9. Predictions about morbidity and mortality were made by the gray system GM(1,1). RESULTS: During the past 10 years, the morbidity and mortality of lung cancer were placed in uptrend slowly. The average morbidity and mortality of lung cancer were 44.75 per 100000 and 41.37 per 100000 respectively, and lung cancer was the first leading cancer for both episode and death of malignant tumors. The proportions of lung cancer were 25.91% and 33.29% for episode and death in all malignant tumors respectively. A half patients with lung cancer was 20-64 years old. Predictive morbidity and mortality of lung cancer would be 47.79/100000 and 44.81/100000 for men and 45.80/100000 and 42.02/100000 for women respectively. CONCLUSIONS: The morbidity and mortality of lung cancer show a slowly increasing trend. Lung cancer is one of main malignant tumors among people of 20-64 years old. The gradually aging population, environmental pollution and individual unhealthy living habits are the important factors of lung cancer increasing.
RESUMO
Epithelial-to-mesenchymal transition (EMT) is a key process associated with tumor progression and metastasis. To define molecular features associated with EMT states, we undertook an integrative approach combining mRNA, miRNA, DNA methylation, and proteomic profiles of 38 cell populations representative of the genomic heterogeneity in lung adenocarcinoma. The resulting data were integrated with functional profiles consisting of cell invasiveness, adhesion, and motility. A subset of cell lines that were readily defined as epithelial or mesenchymal based on their morphology and E-cadherin and vimentin expression elicited distinctive molecular signatures. Other cell populations displayed intermediate/hybrid states of EMT, with mixed epithelial and mesenchymal characteristics. A dominant proteomic feature of aggressive hybrid cell lines was upregulation of cytoskeletal and actin-binding proteins, a signature shared with mesenchymal cell lines. Cytoskeletal reorganization preceded loss of E-cadherin in epithelial cells in which EMT was induced by TGFß. A set of transcripts corresponding to the mesenchymal protein signature enriched in cytoskeletal proteins was found to be predictive of survival in independent datasets of lung adenocarcinomas. Our findings point to an association between cytoskeletal and actin-binding proteins, a mesenchymal or hybrid EMT phenotype and invasive properties of lung adenocarcinomas.
Assuntos
Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Sobrevivência Celular/fisiologia , Células Epiteliais/metabolismo , Transição Epitelial-Mesenquimal/fisiologia , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Adenocarcinoma/genética , Adenocarcinoma de Pulmão , Caderinas/metabolismo , Adesão Celular/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Sobrevivência Celular/genética , Citoesqueleto/metabolismo , Metilação de DNA , Células Epiteliais/patologia , Transição Epitelial-Mesenquimal/genética , Humanos , Neoplasias Pulmonares/genética , MicroRNAs/genética , Proteínas dos Microfilamentos/metabolismo , Proteômica/métodos , Fator de Crescimento Transformador beta/metabolismo , Regulação para Cima , Vimentina/metabolismoRESUMO
OBJECTIVE: To assess the impact of stomach cancer on the Chinese population by epidemiological analysis of its mortality distribution. METHODS: 1990-1992 data on stomach cancer mortality collected by sampling survey involved one tenth of the total Chinese population. RESULTS: The crude mortality rate of stomach cancer in China was 25.2 per 10(5) (32.8 per 10(5) for males and 17.0 per 10(5) for females), which comprised 23.2% of the total cancer deaths from 1990 to 1992, making stomach cancer the leading cause of cancer death. The stomach cancer mortality rate of males was 1.9 times of that of females. The Chinese mortality rates of stomach cancer adjusted by the world population were 40.8 per 10(5) and 18.6 per 10(5) of males and females, which were 4.2-7.9 (of males) and 3.8-8.0 (of females) times of those in the developed countries. Age-adjusted mortality rates of stomach cancer in China have distinct geographical difference: form the lowest 2.5 per 10(5) to the highest 153.0 per 10(5) in the 263 surveyed localities, 15.3 per 10(5) in urban areas and 24.4 per 10(5) in rural areas giving a difference of 1.9 times. CONCLUSION: The prevention and treatment of stomach cancer in China, especially in the countryside and the under-developed areas in the northwest, should be a long-term focus in control of cancers of the digestive system. Urgent measures for prevention and early detection of stomach cancer should be taken.