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1.
Molecules ; 24(22)2019 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-31726781

RESUMO

Citrus junos Tanaka is a traditional medicine for treating coughs, dyspepsia, diabetes, asthma, neuralgia, and inflammatory disorders, and is distributed in Asia, especially in Korea, Japan, and China. This study aimed to use bioactivity-guided fractionation to find therapeutic phytochemicals from C. junos seeds, which can attenuate inflammatory responses. Nine coumarins (1-9) were isolated from the methanolic extract of C. junos seed shells and the inhibitory effects against inflammatory mediators were investigated using murine macrophages. Among the coumarins, compound 3, isogosferol (ISO), more potently attenuated the production of nitric oxide (NO) in lipopolysaccharide (LPS)-induced RAW 264.7 cells. ISO also inhibited the expression of inducible nitric oxide (iNOS) and cyclooxygenase-2 (COX-2) in LPS-stimulated macrophages. Additionally, the phosphorylation of extracellular-regulated kinases (pERK)1/2 was reduced by ISO. We confirmed that ISO attenuated the release of interleukin-1 beta (IL-1ß), which is a central mediator of the inflammatory response. These results demonstrate that ISO from C. junos seed shells may be a potent therapeutic candidate for the treatment of inflammatory diseases.

2.
Chem Biol Interact ; 311: 108755, 2019 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-31319077

RESUMO

Effective control of white adipose tissue accumulation would provide a therapeutic strategy for obesity, which poses a growing global problem. The plant chemical mangiferin stimulates adenosine monophosphate-activated protein kinase (AMPK), which inhibits adipogenesis and has therefore been considered a therapeutic target for obesity and related diseases. We previously reported the anti-inflammatory properties of 6'-O-acetyl mangiferin (OAM). In this study, we evaluated the potential of OAM as an AMPK activator in vitro in 3T3-L1 preadipocytes. OAM inhibited adipogenesis as indicated by lower intracellular lipid and triglyceride accumulation as well as reduced adipogenic gene and protein expression upon treatment. OAM-treated 3T3-L1 cells excreted more glycerol, indicating increased lipolysis, which was supported by increased expression of lipolysis-related genes, including adipose triglyceride lipase and hormone-sensitive lipase. We determined that OAM upregulates lipolysis via phosphorylation-dependent activation of AMPK. Further, OAM upregulated the ß-oxidation pathway as indicated by enhanced expression of phosphorylated acetyl-CoA-carboxylase and long-chain acyl-CoA synthetase 1. In conclusion, OAM markedly decreased intracellular lipid accumulation by enhancing lipolysis via AMPK activation and by upregulating ß-oxidation. Thus, OAM has potential as a drug for the prevention and/or improvement of obesity and related diseases and deserves further study.


Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Adipogenia/efeitos dos fármacos , Iris (Planta)/química , Lipólise/efeitos dos fármacos , Xantonas/farmacologia , Células 3T3-L1 , Proteínas Quinases Ativadas por AMP/genética , Acetil-CoA Carboxilase/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Iris (Planta)/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Camundongos , PPAR gama/genética , PPAR gama/metabolismo , Fosforilação/efeitos dos fármacos , Triglicerídeos/metabolismo
3.
J Neurol ; 266(9): 2286-2293, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31175434

RESUMO

BACKGROUND AND PURPOSE: This study aimed to evaluate the efficacy of intra-arterial thrombectomy (IAT) and prognosis for acute ischaemic stroke patients with active cancer. METHODS: We retrospectively reviewed 253 patients who underwent IAT within 24 h after stroke onset between January 2012 and August 2017. We classified the patients into active cancer (n = 26) and control groups (n = 227) and compared clinical data. Primary outcome was a modified Rankin scale score at 3 months with ordinal logistic regression (shift analysis). RESULTS: Initial National Institutes of Health Stroke Scale (NIHSS) and rate of successful recanalisation did not differ between groups, but the active cancer group showed poor outcomes at 3 months on shift analysis (P = 0.001). The independent predictors of poor prognosis were age [adjusted common odds ratio (aOR) 1.03, 95% confidence interval (CI) 1.01-1.05], baseline NIHSS (aOR 1.14, 95% CI 1.09-1.19), baseline C-reactive protein level (aOR 1.14, 95% CI 1.03-1.25), any cerebral haemorrhage (aOR 1.92, 95% CI 1.21-3.06), and active cancer (aOR 2.35, 95% CI 1.05-5.25). Mortality at 90 days was 30.8% in the cancer group and 8.8% in the control group (P = 0.003). CONCLUSIONS: Although baseline characteristics and recanalisation rate after IAT up to 24 h after stroke onset were similar between acute ischaemic stroke patients with active cancer and without any cancer, stroke-related death and short-term outcome were significantly poorer in patients with active cancer than the controls. Post-procedural haemorrhage and active cancer itself were independent predictors of a decrease in functional independence at 3 months.

4.
Chem Biodivers ; 16(7): e1900033, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30977279

RESUMO

Achillea alpina is widely distributed in Korea and is often used as a folk medicine for stomach disorders. Although a previous study isolated antioxidant compounds (flavonoid O-glucoside, sesquiterpene) from this plant, no systematic study of its chemical constituents had been reported. The present study aimed to identify the phytochemicals present in a methanol extract of A. alpina, assess their potential antioxidant activities in vitro, and determine their effects on melanogenesis in B16F10 melanoma cells. Column chromatographic separation of aqueous fractions of A. alpina led to the isolation of 17 compounds. The chemical structures of these compounds were determined using spectroscopic data from electrospray ionization-mass spectrometry and nuclear magnetic resonance. To the best of our knowledge, the present study is the first to identify compounds 2-10 and 12-17 in A. alpina. Furthermore, compound 6 possessed powerful antioxidant activity, while compound 15 suppressed intracellular tyrosinase activity and thus reduced melanogenesis in B16F10 cells. Therefore, our research suggested that these naturally occurring compounds have the potential to reduce oxidative stress and promote skin whitening. Further investigations will be required to elucidate the mechanisms underlying the antioxidant and antityrosinase activities of these compounds.


Assuntos
Achillea/química , Antineoplásicos Fitogênicos/farmacologia , Antioxidantes/farmacologia , Compostos de Bifenilo/antagonistas & inibidores , Picratos/antagonistas & inibidores , Componentes Aéreos da Planta/química , Animais , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Antioxidantes/química , Antioxidantes/isolamento & purificação , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Camundongos , Estrutura Molecular , Relação Estrutura-Atividade
5.
J Neurol ; 266(3): 766-772, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30673852

RESUMO

The imaging definition of lacunar infarcts is variable, particularly regarding their size and the presence of cavitation. We investigated the changes of diameter and evolution pattern of acute lacunar infarcts, and the factors associated with the evolution pattern. Patients with acute single subcortical hemispheric or brainstem ischemic lesions of penetrating arterial territories were included. Maximal diameters on initial diffusion-weighted image (DWI) and follow-up fluid-attenuated inversion recovery image (FLAIR), which performed > 12 months after initial DWI, were semi-automatically measured. Clinical characteristics were compared according to evolution patterns on follow-up FLAIR, classified as cavitated, focal lesion without cavitation, and disappeared. Five hundred nine patients were included. Mean time to follow-up was 31.3 ± 13.7 months. Mean diameter of acute lacunar lesions decreased from 12.9 ± 4.4 to 8.5 ± 4.8 mm during follow-up. Lesions of 58.2% patients remained as cavitated, 18.3% as focal lesion without cavitation, and 23.6% disappeared. Initial NIHSS score (p = 0.005), diameter of initial lesion (p < 0.001), number of slices showing acute lesion on DWI (p < 0.001), progression of white matter lesion (p < 0.001), number of acute lesions involving gray matter (p = 0.008) and lesion location (p < 0.001) were different among three groups. After adjustment for covariates, diameter of the acute lesion, initial number of old lacunes, and anterior lesion location were associated with the appearance of cavitation. Initial lesion diameter and posterior lesion location were associated with the disappearance. We observed reduction of the acute lacunar lesion diameter in 86%. There were predictive factors of disappearance and cavitation of acute lacunar infarction.


Assuntos
Progressão da Doença , Fibrinolíticos/farmacologia , Imagem por Ressonância Magnética/métodos , Avaliação de Resultados (Cuidados de Saúde) , Índice de Gravidade de Doença , Acidente Vascular Cerebral Lacunar/tratamento farmacológico , Acidente Vascular Cerebral Lacunar/patologia , Idoso , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Acidente Vascular Cerebral Lacunar/diagnóstico por imagem
6.
Mol Cells ; 41(10): 909-916, 2018 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-30165730

RESUMO

In pancreatic ß cells, glucose stimulates the biosynthesis of insulin at transcriptional and post-transcriptional levels. The RNA-binding protein, polypyrimidine tract-binding protein 1 (PTBP1), also named hnRNP I, acts as a critical mediator of insulin biosynthesis through binding to the pyrimidine-rich region in the 3'-untranslated region (UTR) of insulin mRNA. However, the underlying mechanism that regulates its expression in ß cells is unclear. Here, we report that glucose induces the expression of PTBP1 via the insulin receptor (IR) signaling pathway in ß cells. PTBP1 is present in ß cells of both mouse and monkey, where its levels are increased by glucose and insulin, but not by insulin-like growth factor 1. PTBP1 levels in immortalized ß cells established from wild-type (ßIRWT) mice are higher than levels in ß cells established from IR-null (ßIRKO) mice, and ectopic re-expression of IR-WT in ßIRKO cells restored PTBP1 levels. However, PTBP1 levels were not altered in ßIRKO cells transfected with IR-3YA, in which the Tyr1158/1162/1163 residues are substituted with Ala. Consistently, treatment with glucose or insulin elevated PTBP1 levels in ßIRWT cells, but not in ßIRKO cells. In addition, silencing Akt significantly lowered PTBP1 levels. Thus, our results identify insulin as a pivotal mediator of glucose-induced PTBP1 expression in pancreatic ß cells.

7.
J Pathol ; 246(2): 231-243, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30014466

RESUMO

For the majority of patients diagnosed with pancreatic neuroendocrine tumors (NETs), there is significant malignant potential with a poor prognosis; however, the molecular abnormalities and pathogenesis of pancreatic NETs have not been firmly established. Here, we report that loss of expression of the RNA-binding protein HuD correlates with low p27Kip1 (p27) levels and poor prognosis in pancreatic NETs. HuD expression was frequently lost in many human pancreatic NETs, and these pancreatic NETs showed aggressive clinicopathological phenotypes with low p27 levels, increased tumor size, higher World Health Organization grade and pT stage of the tumor, and the presence of angioinvasion. Furthermore, loss of HuD was an independent, progression-free prognostic factor in multivariate survival analysis. However, the level of HuR, a member of the same Hu protein family as HuD, was not significantly correlated with pancreatic NET size and progression. Mechanistically, HuD enhanced p27 mRNA translation by interacting with both the 5'-untranslated region (UTR) and the 3'-UTR of p27 mRNA, and consequently suppressed cell cycle progression and tumor growth. In addition, HuD competed with miR-30a-3p for binding to the 3'-UTR of p27 mRNA, suggesting an interplay between HuD and miR-30a-3p in controlling p27 translation. Our results identify HuD as a pivotal suppressor of pancreatic NET growth, and suggest that HuD has potential value as a prognostic factor of pancreatic NETs. Copyright © 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

8.
Intest Res ; 16(2): 293-298, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29743843

RESUMO

Background/Aims: Inadequate bowel preparation can result in prolonged procedure time and increased missed lesion and complication rates. This prospective study aimed to evaluate bowel preparation quality and identify the predictive factors for inadequate bowel preparation in actual clinical practice. Methods: We included 399 patients who underwent colonoscopy between June 2015 and July 2016. Using the Aronchick bowel preparation scale, we defined a score ≤2 as adequate preparation and a score >2 as inadequate preparation. Results: Mean patient age was 58.38±12.97 years; 60.6% were male. Indications for colonoscopy included screening (69.7%) and surveillance after polyp removal (21.3%). A split-dose regimen was prescribed to 55.4% of patients. The inadequate bowel preparation rate was 28.1%. Overall, the median time between the last bowel preparation agent dose and start of colonoscopy was 5.0 hours (range, 1.5-16.0 hours); that of the adequate group was 5.0 hours (range, 1.5-16.0 hours); and that of the inadequate group was 5 hours (range, 2-23 hours). The mean bowel preparation scale score of the ascending colon (1.94±0.25) was significantly higher than that of other colon segments. On multivariate analysis, elderly age, history of cerebrovascular disease, history of gastrectomy or appendectomy, and total preparation solution uptake <2 L were the independent predictors of inadequate bowel preparation. Conclusions: The inadequate bowel preparation rate was 28.1%. Risk factors included elderly age and history of cerebrovascular disease or abdominal surgery. Patients with these risk factors require special care and education.

9.
Toxicol Res ; 34(2): 133-141, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29686775

RESUMO

Anti-cancer drugs such as cisplatin and doxorubicin are effectively used more than radiotherapy. Cisplatin is a chemotherapeutic drug, used for treatment of various forms of cancer. However, it has side effects such as ototoxicity and nephrotoxicity. Cisplatin-induced nephrotoxicity increases tubular damage and renal dysfunction. Consequently, we investigated the beneficial effect of cynaroside on cisplatin-induced kidney injury using HK-2 cell (human proximal tubule cell line) and an animal model. Results indicated that 10 µM cynaroside diminished cisplatin-induced apoptosis, mitochondrial dysfunction and caspase-3 activation, cisplatin-induced upregulation of caspase-3/MST-1 pathway decreased by treatment of cynaroside in HK-2 cells. To confirm the effect of cynaroside on cisplatin-induced kidney injury in vivo, we used cisplatin exposure animal model (20 mg/kg, balb/c mice, i.p., once a day for 3 days). Renal dysfunction, tubular damage and neutrophilia induced by cisplatin injection were decreased by cynaroside (10 mg/kg, i.p., once a day for 3 days). Results indicated that cynaroside decreased cisplatin-induced kidney injury in vitro and in vivo, and it could be used for improving cisplatin-induced side effects. However, further experiments are required regarding toxicity by high dose cynaroside and caspase-3/MST-1-linked signal transduction in the animal model.

10.
J Microbiol Biotechnol ; 28(6): 1030-1036, 2018 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-29642284

RESUMO

Bacillus strains produce various types of antibiotics, and random mutagenesis has traditionally been used to overproduce these natural metabolites. However, this method leads to the accumulation of unwanted mutations in the genome. Here, we rationally designed a single nucleotide substitution in the degU gene to generate a B. subtilis strain displaying increased plipastatin production in a foreign DNA-free manner. The mutant strain (BS1028u) showed improved antifungal activity against Pythium ultimum. Notably, pps operon deletion in BS1028u resulted in complete loss of antifungal activity, suggesting that the antifungal activity strongly depends on the expression of the pps operon. Quantitative real-time PCR and lacZ assays showed that the point mutation resulted in 2-fold increased pps operon expression, which caused the increase in antifungal activity. Likewise, commercial Bacillus strains can be improved to display higher antifungal activity by rationally designed simple modifications of their genome, rendering them more efficient biocontrol agents.


Assuntos
Antifúngicos/metabolismo , Bacillus subtilis/genética , Bacillus subtilis/metabolismo , Proteínas de Bactérias/genética , Ácidos Graxos/biossíntese , Engenharia Metabólica/métodos , Oligopeptídeos/biossíntese , Peptídeos Cíclicos/biossíntese , Mutação Puntual , Antifúngicos/farmacologia , Ácidos Graxos/farmacologia , Perfilação da Expressão Gênica , Oligopeptídeos/farmacologia , Óperon , Peptídeos Cíclicos/farmacologia , Pythium/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Deleção de Sequência
11.
J Biotechnol ; 266: 50-58, 2018 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-29229542

RESUMO

Chromosome-integrated recombinant protein expression in bacteria has advantages for the stable maintenance of genes without any use of antibiotics during large-scale fermentation. Even though different levels of gene expression were reported, depending upon their chromosomal position in bacterial species, only a limited number of integration sites have been used in B. subtilis. In this study, we randomly integrated the GFP and AprE expression cassettes into the B. subtilis genome to determine integration sites that can produce a high yield of heterologous protein expression. Our mariner transposon-based expression cassette integration system was able to find integration sites, which can produce up to 2.9-fold and 1.5-fold increased expression of intracellular GFP and extracellular AprE, respectively, compared to the common integration site amyE. By analyzing the location of integration sites, we observed an adjacent promoter effect, gene dosage effect, and gene knock-out effect all complexly contributing to the increased level of integrated gene expression. Besides obtaining a high yield of heterologous protein expression, our system can also provide a wide-range of expression to expand the systematic application for steady-state metabolic protein production.


Assuntos
Bacillus subtilis/genética , Elementos de DNA Transponíveis , Expressão Gênica , Técnicas de Introdução de Genes/métodos , Bacillus subtilis/metabolismo , Proteínas de Fluorescência Verde/biossíntese , Proteínas de Fluorescência Verde/economia , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética
12.
Korean J Gastroenterol ; 72(5): 258-261, 2018 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-30642141

RESUMO

Esophageal basaloid squamous carcinoma (BSC) is a rare, aggressive variant of squamous cell carcinoma. BSC is usually diagnosed in advanced stage and its prognosis is relatively poor. A 59-year-old male with subepithelial lesion of the esophagus that was incidentally discovered during health promotion examination was referred to our hospital. Esophagogastroduodenoscopy showed a 10-mm bulging mucosa with an intact surface at 34 cm from incisor teeth. Endoscopic ultrasonography revealed a smooth margined homogenous hypoechoic lesion, measuring 11.3×3.9 mm with a submucosal layer of origin. The patient underwent endoscopic mucosal resection of the subepithelial lesion. Pathologic examination of the resected specimen revealed BSC with involvement of vertical margin by tumor. The patient then underwent radiotherapy, and is doing well without recurrence for 35 months. We report a case of esophageal BSC confined to submucosal layer successfully treated with endoscopic resection followed by radiation.


Assuntos
Neoplasias Esofágicas/diagnóstico , Carcinoma de Células Escamosas do Esôfago/diagnóstico , Endossonografia , Neoplasias Esofágicas/patologia , Neoplasias Esofágicas/radioterapia , Carcinoma de Células Escamosas do Esôfago/patologia , Carcinoma de Células Escamosas do Esôfago/radioterapia , Esofagoscopia , Esôfago/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias
13.
Stem Cells Int ; 2017: 8085462, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28588623

RESUMO

Previously, the perivascular characteristics of dental pulp stem cells (DPSCs) were reported, which suggested the potential application of DPSCs as perivascular cell source. In this study, we investigated whether DPSCs had angiogenic capacity by coinjection with human umbilical vein endothelial cells (HUVECs) in vivo; in addition, we determined the role of stromal cell-derived factor 1-α (SDF-1α) and C-X-C chemokine receptor type 4 (CXCR4) axis in the mutual interaction between DPSCs and HUVECs. Primarily isolated DPSCs showed mesenchymal stem cell- (MSC-) like characteristics. Moreover, DPSCs expressed perivascular markers such as NG2, α-smooth muscle actin (α-SMA), platelet-derived growth factor receptor ß (PDGFRß), and CD146. In vivo angiogenic capacity of DPSCs was demonstrated by in vivo Matrigel plug assay. We could observe microvessel-like structures in the coinjection of DPSCs and HUVECs at 7 days postinjection. To block SDF-1α and CXCR4 axis between DPSCs and HUVECs, AMD3100, a CXCR4 antagonist, was added into Matrigel plug. No significant microvessel-like structures were observed at 7 days postinjection. In conclusion, DPSCs have perivascular characteristics that contribute to in vivo angiogenesis. The findings of this study have potential applications in neovascularization of engineered tissues and vascular diseases.

14.
Oncol Lett ; 14(6): 8213-8219, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29344264

RESUMO

The promoter region of the telomerase reverse transcriptase gene (TERT) is mutated in a subpopulation of patients with glioblastoma multiforme (GBM). In the present study, preclinical and clinical implications of the mutation were analyzed in 25 GBMs to evaluate its utility as a therapeutic target. Associations between the TERT promoter mutation and a number of preclinical/clinical characteristics were analyzed. Notably, the TERT promoter mutation was identified in 92.3% of GBMs where dissociated cells revealed in vitro sphere formation capacity; while the TERT promoter mutation was identified in 33.3% of GBMs without in vitro sphere formation capacity (P=0.004). In addition, this significantly increased mutation rate was observed in GBMs with in vivo tumorigenic potential (80% vs. 0%; P=0.004). Furthermore, patients with GBM exhibiting the TERT promoter mutation demonstrated significantly decreased overall survival rate compared with patients lacking this mutation (81.7 vs. 152.6 weeks; P=0.026). The results of the present study indicated that the TERT promoter mutation is associated with the self-renewal capacity of GBM cells and clinical aggressiveness of GBMs, which may be translated to a targeting therapy against TERT to inhibit the self-renewal of GBM cells.

15.
Food Sci Biotechnol ; 26(5): 1209-1215, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-30263654

RESUMO

Five proanthocyanidins, two B-type dimers and three A-type trimers, were purified and isolated from the fruit peels of Pyrus pyrifolia Nakai cv. Chuhwangbae. The isolated compounds were identified as (-)-epicatechin gallate-(4ߠ→ 8)-(-)-epicatechin (Hahashi et al. in Ann Biol Res 3:3200-3207, 2012), (-)-epicatechin-(4ߠ→ 8)-(-)-epicatechin (procyanidin B2) (Tanrioven and Eksi in Food Chem 93:89-93, 2005), (-)-epicatechin-(4ߠ→ 8, 2ߠ→ O-7)-(-)-epicatechin-(4ߠ→ 8)-(-)-epicatechin (cinnamtannins B1) (Salta et al. in J. Fun. Food 2: 153-157, 2010), (-)-epicatechin-(4ߠ→ 8)-(-)-epicatechin-(4ߠ→ 8, 2ߠ→ O-7)-(-)-epicatechin (aesculitannin A) (Challice and Westwood in Phytochemistry 11: 37-44, 1972), and (-)-epicatechin-(4ߠ→ 6)-(-)-epicatechin-(4ߠ→ 8, 2ߠ→ O→7)-(-)-epicatechin (Es-Safi et al. in J Agric Food Chem 54: 6969-6977, 2006). Their structures were determined by nuclear magnetic resonance and mass spectrometry. The three A-type proanthocyanidin trimers were identified for the first time from pear.

16.
Oncotarget ; 7(32): 51626-51639, 2016 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-27438149

RESUMO

Muscle-invasive bladder cancer (MIBC) consists of a heterogeneous group of tumors with a high rate of metastasis and mortality. To facilitate the in-depth investigation and validation of tailored strategies for MIBC treatment, we have developed an integrated approach using advanced high-throughput drug screening and a clinically relevant patient-derived preclinical platform. We isolated patient-derived tumor cells (PDCs) from a rare MIBC case (BD-138T) that harbors concomitant epidermal growth factor receptor (EGFR) amplification and phosphatase and tensin homolog (PTEN) deletion. High-throughput in vitro drug screening demonstrated that dasatinib, a SRC inhibitor, and PKI-587, a dual PI3K/mTOR inhibitor, exhibited targeted anti-proliferative and pro-apoptotic effects against BD-138T PDCs. Using established patient-derived xenograft models that successfully retain the genomic and molecular characteristics of the parental tumor, we confirmed that these anti-tumor responses occurred through the inhibition of SRC and PI3K/AKT/mTOR signaling pathways. Taken together, these experimental results demonstrate that dasatinib and PKI-587 might serve as promising anticancer drug candidates for treating MIBC with combined EGFR gene amplification and PTEN deletion.


Assuntos
Antineoplásicos/farmacologia , Carcinoma de Células de Transição/patologia , Dasatinibe/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Receptores ErbB/genética , PTEN Fosfo-Hidrolase/genética , Neoplasias da Bexiga Urinária/patologia , Antineoplásicos/uso terapêutico , Carcinoma de Células de Transição/tratamento farmacológico , Carcinoma de Células de Transição/genética , Linhagem Celular Tumoral , Dasatinibe/uso terapêutico , Amplificação de Genes , Deleção de Genes , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Musculares/tratamento farmacológico , Neoplasias Musculares/genética , Neoplasias Musculares/secundário , Mutação , Invasividade Neoplásica , Cultura Primária de Células/métodos , Neoplasias da Bexiga Urinária/tratamento farmacológico , Neoplasias da Bexiga Urinária/genética
17.
PLoS One ; 11(7): e0158639, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27391353

RESUMO

Stem cells and therapeutic genes are emerging as a new therapeutic approach to treat various neurodegenerative diseases with few effective treatment options. However, potential formation of tumors by stem cells has hampered their clinical application. Moreover, adequate preclinical platforms to precisely test tumorigenic potential of stem cells are controversial. In this study, we compared the sensitivity of various animal models for in vivo stem cell tumorigenicity testing to identify the most sensitive platform. Then, tumorigenic potential of adult human multipotent neural cells (ahMNCs) immortalized by the human telomerase reverse transcriptase (hTERT) gene was examined as a stem cell model with therapeutic genes. When human glioblastoma (GBM) cells were injected into adult (4-6-week-old) Balb/c-nu, adult NOD/SCID, adult NOG, or neonate (1-2-week-old) NOG mice, the neonate NOG mice showed significantly faster tumorigenesis than that of the other groups regardless of intracranial or subcutaneous injection route. Two kinds of ahMNCs (682TL and 779TL) were primary cultured from surgical samples of patients with temporal lobe epilepsy. Although the ahMNCs were immortalized by lentiviral hTERT gene delivery (hTERT-682TL and hTERT-779TL), they did not form any detectable masses, even in the most sensitive neonate NOG mouse platform. Moreover, the hTERT-ahMNCs had no gross chromosomal abnormalities on a karyotype analysis. Taken together, our data suggest that neonate NOG mice could be a sensitive animal platform to test tumorigenic potential of stem cell therapeutics and that ahMNCs could be a genetically stable stem cell source with little tumorigenic activity to develop regenerative treatments for neurodegenerative diseases.


Assuntos
Células-Tronco Multipotentes/citologia , Células-Tronco Multipotentes/metabolismo , Telomerase/metabolismo , Adulto , Animais , Carcinogênese/genética , Diferenciação Celular/genética , Diferenciação Celular/fisiologia , Células Cultivadas , Humanos , Imuno-Histoquímica , Cariótipo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Telomerase/genética , Telômero/genética , Adulto Jovem
18.
Medicine (Baltimore) ; 95(26): e4028, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27368026

RESUMO

To avoid missing events associated with clinical activity, the authors previously developed a novel, web-based, self-reporting Crohn disease (CD) symptom diary. However, although this diary provided a means of self-checking based on responses to set questions based on Harvey-Bradshaw index scores, it was limited in terms of describing other specific symptoms. Thus, the authors added a space to the questionnaire, which allows patients to send clinicians questions or a description of unpredictable events. The aim of the present study was to assess the clinical usefulness of this messaging system by analyzing patients' messages.The messaging system between patients and their doctors was included in a webpage created for recording patients' symptom diaries (www.cdsd.or.kr). Using this system, patients can send messages easily at any time and doctors can read and respond to these messages immediately using a smart phone or computer. In the present study, the authors retrospectively reviewed 686 messages sent by 152 patients from July 2012 to July 2014 and patient medical records.Mean patient age was 29.0 ±â€Š11.6 years and the male-to-female ratio was 99:53. Most messages regarded symptoms (381 messages, 55.5%), which was followed by self-reports about general condition (195 messages, 28.4%) and questions about treatment (71 messages, 10.3%). With respect to symptoms, abdominal pain was most common (145 cases, 21.1%) followed by hematochezia (36 cases, 5.2%). Problems about medication were the most frequently associated with treatment (65, 91.5%). Patients above 40 years showed a greater tendency to focus on symptoms and treatment (P = 0.025). The doctor answer rate was 56.3% (n = 386), and based on these responses, an early visit was needed in 28 cases (7.3%).Using this web-based messaging system, patients were able to obtain proper advice from their physicians without visiting clinics or searching the Internet, and in addition, 7.3% of messages prompted an early visit. Although longer follow-up is required, this study shows that the devised messaging system provides a clinically relevant communication tool for patients and physicians.


Assuntos
Doença de Crohn , Internet , Mensagem de Texto , Adulto , Doença de Crohn/terapia , Feminino , Humanos , Masculino , Relações Médico-Paciente , Estudos Retrospectivos
19.
Genome Biol ; 17: 80, 2016 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-27139883

RESUMO

BACKGROUND: Intratumoral heterogeneity hampers the success of marker-based anticancer treatment because the targeted therapy may eliminate a specific subpopulation of tumor cells while leaving others unharmed. Accordingly, a rational strategy minimizing survival of the drug-resistant subpopulation is essential to achieve long-term therapeutic efficacy. RESULTS: Using single-cell RNA sequencing (RNA-seq), we examine the intratumoral heterogeneity of a pair of primary renal cell carcinoma and its lung metastasis. Activation of drug target pathways demonstrates considerable variability between the primary and metastatic sites, as well as among individual cancer cells within each site. Based on the prediction of multiple drug target pathway activation, we derive a combinatorial regimen co-targeting two mutually exclusive pathways for the metastatic cancer cells. This combinatorial strategy shows significant increase in the treatment efficacy over monotherapy in the experimental validation using patient-derived xenograft platforms in vitro and in vivo. CONCLUSIONS: Our findings demonstrate the investigational application of single-cell RNA-seq in the design of an anticancer regimen. The approach may overcome intratumoral heterogeneity which hampers the success of precision medicine.


Assuntos
Carcinoma de Células Renais/tratamento farmacológico , Neoplasias Renais/tratamento farmacológico , Terapia de Alvo Molecular/métodos , Análise de Sequência de RNA/métodos , Análise de Célula Única/métodos , Adulto , Animais , Carcinoma de Células Renais/genética , Carcinoma de Células Renais/patologia , Células Cultivadas , Perfilação da Expressão Gênica/métodos , Humanos , Neoplasias Renais/genética , Neoplasias Renais/patologia , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/secundário , Masculino , Camundongos
20.
Oncotarget ; 6(32): 33046-64, 2015 10 20.
Artigo em Inglês | MEDLINE | ID: mdl-26426994

RESUMO

Despite advances in the development of molecularly targeted therapies, metastatic renal cell carcinoma (RCC) is still incurable. Artesunate (ART), a well-known anti-malarial drug with low toxicity, exhibits highly selective anti-tumor actions against various tumors through generation of cytotoxic carbon-centered free radical in the presence of free iron. However, the therapeutic efficacy of ART against metastatic RCC has not yet been fully elucidated. In the analysis on a dataset from The Cancer Genome Atlas (TCGA) (n = 469) and a tissue microarray set from Samsung Medical Center (n = 119) from a cohort of patients with clear cell RCC (ccRCC), up-regulation of transferrin receptor 1 (TfR1), which is a well-known predictive marker for ART, was correlated with the presence of distant metastasis and an unfavorable prognosis. Moreover, ART exerted potent selective cytotoxicity against human RCC cell lines (Caki-1, 786-O, and SN12C-GFP-SRLu2) and sensitized these cells to sorafenib in vitro, and the extent of ART cytotoxicity correlated with TfR1 expression. ART-mediated growth inhibition of human RCC cell lines was shown to result from the induction of cell cycle arrest at the G2/M phase and oncosis-like cell death. Furthermore, ART inhibited cell clonogenicity and invasion of human RCC cells and anti-angiogenic effects in vitro in a dose-dependent manner. Consistent with these in vitro data, anti-tumor, anti-metastatic and anti-angiogenic effects of ART were also validated in human 786-O xenografts. Taken together, ART is a promising novel candidate for treating human RCC, either alone or in combination with other therapies.


Assuntos
Antineoplásicos/farmacologia , Artemisininas/farmacologia , Carcinoma de Células Renais/tratamento farmacológico , Neoplasias Renais/tratamento farmacológico , Animais , Antígenos CD/metabolismo , Antimaláricos/farmacologia , Artesunato , Carcinoma de Células Renais/irrigação sanguínea , Carcinoma de Células Renais/metabolismo , Carcinoma de Células Renais/patologia , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Feminino , Células Endoteliais da Veia Umbilical Humana , Humanos , Neoplasias Renais/irrigação sanguínea , Neoplasias Renais/metabolismo , Neoplasias Renais/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Invasividade Neoplásica , Metástase Neoplásica , Neovascularização Patológica/tratamento farmacológico , Prognóstico , Espécies Reativas de Oxigênio/metabolismo , Receptores da Transferrina/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
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