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2.
Oncogene ; 40(30): 4847-4858, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34155349

RESUMO

Small cell lung cancer (SCLC) continues to cause poor clinical outcomes due to limited advances in sustained treatments for rapid cancer cell proliferation and progression. The transcriptional factor Forkhead Box M1 (FOXM1) regulates cell proliferation, tumor initiation, and progression in multiple cancer types. However, its biological function and clinical significance in SCLC remain unestablished. Analysis of the Cancer Cell Line Encyclopedia and SCLC datasets in the present study disclosed significant upregulation of FOXM1 mRNA in SCLC cell lines and tissues. Gene set enrichment analysis (GSEA) revealed that FOXM1 is positively correlated with pathways regulating cell proliferation and DNA damage repair, as evident from sensitization of FOXM1-depleted SCLC cells to chemotherapy. Furthermore, Foxm1 knockout inhibited SCLC formation in the Rb1fl/flTrp53fl/flMycLSL/LSL (RPM) mouse model associated with increased levels of neuroendocrine markers, Ascl1 and Cgrp, and decrease in Yap1. Consistently, FOXM1 depletion in NCI-H1688 SCLC cells reduced migration and enhanced apoptosis and sensitivity to cisplatin and etoposide. SCLC with high FOXM1 expression (N = 30, 57.7%) was significantly correlated with advanced clinical stage, extrathoracic metastases, and decrease in overall survival (OS), compared with the low-FOXM1 group (7.90 vs. 12.46 months). Moreover, the high-FOXM1 group showed shorter progression-free survival after standard chemotherapy, compared with the low-FOXM1 group (3.90 vs. 8.69 months). Our collective findings support the utility of FOXM1 as a prognostic biomarker and potential molecular target for SCLC.


Assuntos
Biomarcadores Tumorais , Proteína Forkhead Box M1/genética , Neoplasias Pulmonares/etiologia , Neoplasias Pulmonares/mortalidade , Carcinoma de Pequenas Células do Pulmão/etiologia , Carcinoma de Pequenas Células do Pulmão/mortalidade , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Linhagem Celular Tumoral , Proliferação de Células , Modelos Animais de Doenças , Feminino , Proteína Forkhead Box M1/metabolismo , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Neoplasias Pulmonares/diagnóstico , Masculino , Camundongos , Camundongos Transgênicos , Pessoa de Meia-Idade , Gradação de Tumores , Estadiamento de Neoplasias , Prognóstico , Carcinoma de Pequenas Células do Pulmão/diagnóstico , Microtomografia por Raio-X , Ensaios Antitumorais Modelo de Xenoenxerto
3.
Int J Nanomedicine ; 15: 8411-8426, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33149584

RESUMO

BACKGROUND: Gold nanoparticles with high biocompatibility and immunomodulatory properties have potential applications in the development of new diagnostic and therapeutic strategies for nanomedicine. Nanoparticles targeting macrophages can manipulate or control immunological diseases. This study assessed the activity of dendrimer-encapsulated gold nanodots (AuNDs) with three surface modifications [ie, outfacing groups with primary amine (AuNDs-NH2), hydroxyl (AuNDs-OH), and quaternary ammonium ions (AuNDs-CH3)] regulated macrophage function and antioxidant response through Nrf2-dependent pathway. METHODS: AuNDs were prepared and characterized. Intracellular distribution of AuNDs in human macrophages was observed through confocal microscopy. The activity of AuNDs was evaluated using macrophage functions and antioxidant response in the human macrophage cell line THP-1. RESULTS: AuNDs-NH2 and AuNDs-CH3, but not AuNDs-OH, drove the obvious Nrf2-antioxidant response element pathway in THP-1 cells. Of the three, AuNDs-NH2 considerably increased mRNA levels and antioxidant activities of heme oxygenase 1 and NAD(P)H quinone dehydrogenase 1 in THP-1 cells. IL-6 mRNA and protein expression was mediated through Nrf2 activation in AuNDs-NH2-treated macrophages. Furthermore, Nrf2 activation by AuNDs-NH2 increased the phagocytic ability of THP-1 macrophages. CONCLUSION: AuNDs-NH2 had immunomodulatory activities in macrophages. The findings of the present work suggested that AuNDs have potential effects against chronic inflammatory diseases via the Nrf2 pathway.


Assuntos
Aminas/química , Antioxidantes/metabolismo , Ouro/química , Macrófagos/metabolismo , Nanopartículas Metálicas/química , Terapia de Alvo Molecular , Fator 2 Relacionado a NF-E2/metabolismo , Citocinas/metabolismo , Endotoxinas/metabolismo , Heme Oxigenase-1/metabolismo , Humanos , Mediadores da Inflamação/metabolismo , Estresse Oxidativo , Fagocitose , Fosforilação , Espécies Reativas de Oxigênio/metabolismo , Células THP-1
4.
Environ Pollut ; 267: 115548, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32892025

RESUMO

The level of di-(2-ethylhexyl) phthalate (DEHP) is elevated in chronic kidney disease patients undergoing dialysis. However, statins are unable to reduce the cardiovascular events in chronic dialysis patients. In this study, we investigated the effects of DEHP on statin-conferred pleiotropic effects and the underlying molecular mechanism in peritoneal dialysis (PD) patients and endothelial cells (ECs). In PD patients with serum DEHP level ≥0.0687 µg/mL, statin treatment was not associated with lower risk of cardiovascular disease. In ECs, exposure to DEHP abrogated the simvastatin-induced NO bioavailability and EC-related functions. Additionally, DEHP abolished the anti-inflammatory effect of simvastatin on the tumor necrosis factor α-induced upregulation of adhesion molecules and monocyte adhesion to ECs. Mechanistically, DEHP blunted the activation of transient receptor potential vanilloid type 1 (TRPV1), which is required for NO production by simvastatin in ECs. Notably, DEHP increased the activity and expression of protein phosphatase 2B (PP2B), a negative regulator of TRPV1 activity. The effect of DEHP on PP2B activation was mediated by the activation of the NADPH oxidase/reactive oxygen species (NOX-ROS) pathway. Inhibition of PP2B activity by pharmacological antagonists prevented the inhibitory effects of DEHP on simvastatin-induced Ca2+ influx, NO bioavailability, and EC migration, proliferation, tube formation, and anti-inflammatory action. Collectively, DEHP activates the NOX-ROS-PP2B pathway, which in turns inhibits TRPV1/Ca2+-dependent signaling and abrogates the statin-conferred pleiotropic protection in ECs.


Assuntos
Dietilexilftalato , Inibidores de Hidroximetilglutaril-CoA Redutases , Insuficiência Renal Crônica , Dietilexilftalato/toxicidade , Células Endoteliais , Humanos , Ácidos Ftálicos , Diálise Renal , Insuficiência Renal Crônica/terapia
5.
Toxicol In Vitro ; 59: 246-254, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31009676

RESUMO

Previous studies revealed that cellular accumulation of mono(2-ethylhexyl)phthalate (MEHP) disturbed energy metabolism in adipocytes, where glucose uptake was significantly increased. The present study aimed to determine the mechanisms underlying the increased glucose uptake. MEHP-treated 3T3-L1 adipocytes exhibited a significantly increased glucose uptake activity. Immunoblot analysis suggested that the insulin-induced signals were not responsible for the increased glucose uptake. qPCR analysis revealed that both Glut1 and Glut4 genes were highly expressed during adipogenesis; Glut1 mRNA levels in MEHP-treated adipocytes were significantly increased. Moreover, MEHP-treated adipocytes exhibited significantly increased levels of fibroblast growth factor 21 (FGF21) in both mRNA and secreted protein. FGF21 is a peptide hormone with pleiotropic effects on regulation of insulin sensitivity and glucose/lipid homeostasis. We found that MEHP, FGF21, and lactate in culture medium together enhanced Fgf21 gene expression in MEHP-treated adipocytes. FGF21 signaling requires fibroblast growth factor receptor (FGFR) and ßKlotho. Fgfr family and ßKlotho genes were actively expressed during adipogenesis; mRNA levels of Fgfr3 and Fgfr4 genes in MEHP-treated adipocytes were significantly increased. Roles of FGF21/FGFR and phosphoinositide 3-kinase (PI3K)/AKT signal axes in regulation of glucose uptake were determined. We demonstrated that FGF21/FGFR signals played the major roles in up-regulation of the basal glucose uptake in MEHP-treated adipocytes. The in vitro evidence suggests that cellular FGF21 secretion enhances the basal glucose uptake in MEHP-treated adipocytes.


Assuntos
Células 3T3-L1/efeitos dos fármacos , Adipócitos/efeitos dos fármacos , Dietilexilftalato/análogos & derivados , Fatores de Crescimento de Fibroblastos/metabolismo , Glucose/metabolismo , Células 3T3-L1/metabolismo , Adipócitos/metabolismo , Animais , Dietilexilftalato/farmacologia , Insulina/metabolismo , Camundongos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo
6.
Environ Sci Technol ; 51(24): 14262-14272, 2017 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-29192765

RESUMO

Polycyclic aromatic hydrocarbons (PAHs) are widely distributed throughout the atmosphere as mixtures attached to ambient particulate matter (PM). PAHs usually elicit similar toxicological pathways but do so with varying levels of efficacy. In this study, we utilized high-throughput screening (HTS) in vitro data of PAHs to predict health risks associated with coarse and fine PM. PM samples with 22 PAH compounds obtained from residential areas close to industrial parks in central Taiwan were analyzed. On the basis of the PM-bound PAH concentrations and their activities reported in HTS assays, we developed a probabilistic model for estimating cumulative exposure of humans to PAHs. Activity-to-exposure ratio (AER) values were calculated to compare relative risks of activating the aryl hydrocarbon receptor (AhR), nuclear factor erythroid 2-related factor 2 (Nrf2), and tumor suppressor gene (p53) when children or adults were exposed to fine or coarse PM in different seasons. On the basis of AER values, the risk of fine PM exposure was relatively higher than the risk of exposure to coarse PM in pathway activation. Children as a susceptible population had a risk of the activating AhR pathway greater than that of adults. Particularly higher risks were observed in winter than in summer. Among three pathways, AhR was the most sensitive one activated by exposure to PAHs. In addition, the activation of the AhR, Nrf2, and p53 pathways was compared by in vitro reporter assays with and without the pre-extraction of PAHs from PM. Our proposed novel approach accounts for mixture toxicities in characterizing in vitro pathway-based risks via inhalation exposure to ambient PAHs.


Assuntos
Material Particulado , Hidrocarbonetos Policíclicos Aromáticos , Medição de Risco , Poluentes Atmosféricos , Humanos , Estações do Ano , Taiwan
7.
Sci Rep ; 7(1): 15320, 2017 11 10.
Artigo em Inglês | MEDLINE | ID: mdl-29127306

RESUMO

Nickel compounds have been classified as carcinogens and shown to be associated with induction of epithelial-mesenchymal transition (EMT) in fibrogenesis and tumorigenesis, as well as the crucial role of microRNAs (miRNAs) and their related genes in controlling EMT and cancer metastasis. Thus, the mechanisms involved in the regulation of EMT in nickel-treated cells are of potential interest in understanding lung fibrosis and tumor progression. We investigated the miRNA-dependent mechanisms involved in nickel-induced EMT in lung epithelial cells. Nickel increased miR-4417 expression and decreased its target gene TAB2 expression. Treatment of cells with TGF-ß inhibitor SB525334 significantly blocked NiCl2 and TGF-ß-induced EMT. The expression of miR-4417 was abolished by SB525334 in TGF-ß-treated cells, but not in nickel-treated cells. Both overexpression of miR-4417 and silencing of TAB2 induced fibronectin expression, but did not reduce E-cadherin expression. Moreover, oral administration of nickel promoted lung tumor growth in nude mice that had received BEAS-2B transformed cells by intravenous injection. The induction of EMT by nickel is mediated through multiple pathways. Induction of abundant miR-4417 and reduction of TAB2 expression following nickel exposure and may be involved in nickel-induced fibronectin. These findings provide novel insight into the roles of nickel in fibrogenesis and tumor progression.


Assuntos
Transformação Celular Neoplásica/metabolismo , Células Epiteliais/metabolismo , Neoplasias Pulmonares/metabolismo , Pulmão/metabolismo , MicroRNAs/metabolismo , Níquel/toxicidade , Fibrose Pulmonar/metabolismo , RNA Neoplásico/metabolismo , Mucosa Respiratória/metabolismo , Linhagem Celular Transformada , Transformação Celular Neoplásica/induzido quimicamente , Transformação Celular Neoplásica/patologia , Células Epiteliais/patologia , Humanos , Pulmão/patologia , Neoplasias Pulmonares/induzido quimicamente , Neoplasias Pulmonares/patologia , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/patologia , Mucosa Respiratória/patologia
8.
J Appl Toxicol ; 37(12): 1537-1546, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28849599

RESUMO

The study aimed to examine effects of environmental estrogens at body burden levels on energy metabolism in fat cells. Acclimation of T47D-KBluc cells in estrogen-deprived medium was established for high performance of estrogen-responsive luciferase reporter assay. With the assay, relative estrogenic potency of four selected estrogen receptor (ER) agonists, i.e. diethylstilbestrol, ß-estradiol, 4-nonylphenol and bisphenol A, were determined. Immunoblot analysis revealed that the ER agonists at both EC80 and EC100 caused rapid and transient phosphorylation of extracellular signal-regulated kinases (ERK) in an ER-dependent manner. 3T3-L1 adipocytes treated with the ER agonists at EC80 for 24 hours exhibited significant downregulation in mitochondrial respiration and glycolytic function. Importantly, EC80 values of 4-nonylphenol (6.0 × 10-10  m) and bisphenol A (1.0 × 10-8  m) are in the range of human body burdens. The finding that estrogenic chemicals at body burden levels cause significant impact on fat cell energy metabolism raises an important public health issue that deserves more attention.


Assuntos
Adipócitos/efeitos dos fármacos , Adipogenia/efeitos dos fármacos , Metabolismo Energético/efeitos dos fármacos , Estrogênios não Esteroides/toxicidade , Estrogênios/toxicidade , Receptores de Estrogênio/metabolismo , Células 3T3-L1 , Adipócitos/metabolismo , Animais , Carga Corporal (Radioterapia) , Relação Dose-Resposta a Droga , Camundongos
9.
Oncotarget ; 7(31): 49310-49321, 2016 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-27384479

RESUMO

The development of resistance to platinum drugs in cancer cells severely reduces the efficacy of these drugs. Thus, the discovery of novel drugs or combined strategies to overcome drug resistance is imperative. In addition to our previous finding that combined D-penicillamine with platinum drugs exerts synergistic cytotoxicity, we recently identified a novel therapeutic strategy by combining an iron chelating agent desferal with platinum drugs to overcome platinum resistance in an oxaliplatin-resistant human cervical cancer cell line, S3. Further study demonstrated that the level of platinum-DNA adduct formation positively correlated with cell death in combination of desferal with platinums than that of each drug alone in S3 cells. Decrement of human copper transporter 1 (hCtr1) and transferrin receptor 1 (TfR1) expression involved in the development of platinum resistance in S3 cells. Moreover, desferal promoted the expression of hCtr1 through the upregulation of Sp1. The overexpression of Sp1 increased the expression of NF-κB and translocated it into the nucleus to bind to the TfR1 promoter region, which subsequently increased the expression of TfR1. Importantly, the cotreatment of oxaliplatin with desferal significantly potentiated the oxaliplatin-elicited antitumoral effect in the oxaliplatin-resistant xenograft animal model without any toxic effect observed. Taken together, these results demonstrated that the combination of desferal with oxaliplatin can overcome oxaliplatin resistance through the regulation of hCtr1 and TfR1, and may have beneficial effect for treatment of patient with oxaliplatin-refractory tumors.


Assuntos
Antígenos CD/metabolismo , Proteínas de Transporte de Cátions/metabolismo , Desferroxamina/administração & dosagem , Compostos Organoplatínicos/administração & dosagem , Receptores da Transferrina/metabolismo , Fator de Transcrição Sp1/metabolismo , Neoplasias do Colo do Útero/tratamento farmacológico , Transporte Ativo do Núcleo Celular , Animais , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Núcleo Celular/metabolismo , Quelantes/química , Transportador de Cobre 1 , Adutos de DNA , Resistencia a Medicamentos Antineoplásicos , Feminino , Humanos , Ferro/química , Masculino , Camundongos , Camundongos Nus , Transplante de Neoplasias , Oxaliplatina , Regiões Promotoras Genéticas , Regulação para Cima
10.
Environ Sci Pollut Res Int ; 23(9): 8518-28, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26791027

RESUMO

Estrogen-like endocrine disrupting compounds (EEDC) such as bisphenol A, nonylphenol, and phthalic acid esters are toxic compounds that may occur in both raw- and drinking water. The aim of this study was to combine chemical- and bioassay to evaluate the risk of EEDCs in the drinking water treatment plants (DWTPs). Fifty-six samples were collected from seven DWTPs located in northern-, central-, and southern Taiwan from 2011 to 2012 and subjected to chemical analyses and two bioassay methods for total estrogenic activity (E-Screen and T47D-KBluc assay). Among of the considered EEDCs, only dibutyl phthalate (DBP) and di (2-ethylhexyl) phthalate (DEHP) were detected in both drinking and raw water samples. DBP levels in drinking water ranged from

Assuntos
Água Potável/química , Exposição Ambiental/estatística & dados numéricos , Estrogênios/análise , Águas Residuárias/química , Poluentes Químicos da Água/análise , Purificação da Água , Compostos Benzidrílicos/análise , Bioensaio , Dibutilftalato/análise , Disruptores Endócrinos/análise , Estradiol/análise , Feminino , Humanos , Masculino , Fenóis/análise , Ácidos Ftálicos/análise , Taiwan
11.
Biochem Pharmacol ; 95(1): 28-37, 2015 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-25801007

RESUMO

The platinum-based regimen is the front-line treatment of chemotherapy. However, development of platinum resistance often causes therapeutic failure in this disease. We previously have generated an oxaliplatin-resistant subline, named S3, from human cervical carcinoma SiHa cells, and its resistant phenotype was well-characterized. In the present study, we aimed to identify the novel therapeutic strategy by combining copper chelator D-penicillamine with oxaliplatin, and to elucidate the underlying mechanisms for overcoming oxaliplatin resistance. As the result, D-penicillamine exerted synergistic killing effects only in S3 cells when combined with oxaliplatin and cisplatin by using Chou-Talalay method. Further study showed that the amounts of platinum DNA adduct formed were positively correlated to the percentage of cell death in S3 cells when co-treated D-penicillamine with oxaliplatin and cisplatin. D-penicillamine promoted copper influx transporter hCtr1 expression through upregulation of Sp1. Sp1 overexpression induced p53 translocation from nucleus to cytosol and caused p53 degradation through ubiquitination, which subsequently suppressed the expression of the copper efflux transporter ATP7A. Importantly, co-treatment of cisplatin with D-penicillamine enhanced oxaliplatin-elicited antitumor effect in the oxalipatin-resistant S3 xenograft tumors, but not found in SiHa xenograft model. Notably, Mice received D-penicillamine alone or in combination of D-penicillamine ad oxalipatin, increased hCtrl protein level in S3 xenograft tumor, however, the protein level of ATP7A was decreased. Taken together, this study provides insight into that the co-manipulation of hCtrl and ATP7A by D-penicillamine could increase the therapeutic efficacy of platinum drugs in oxaliplatin resistant tumors, especially in resistant phenotype with downexpression of hCtrl and overexpression of ATP7A.


Assuntos
Antineoplásicos/administração & dosagem , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Compostos Organoplatínicos/administração & dosagem , Penicilamina/administração & dosagem , Neoplasias do Colo do Útero/tratamento farmacológico , Animais , Quelantes/administração & dosagem , Cisplatino/administração & dosagem , Sinergismo Farmacológico , Quimioterapia Combinada , Feminino , Humanos , Masculino , Camundongos , Camundongos Nus , Oxaliplatina , Neoplasias do Colo do Útero/patologia
12.
Int J Cancer ; 137(6): 1291-305, 2015 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-25693518

RESUMO

Cisplatin (CDDP) is an important anti-cancer drug commonly used in various human cancers, including nasopharyngeal carcinoma (NPC). How to overcome the drug resistance of CDDP provides opportunities to improve clinical outcomes of NPC. O(6) -methylguanine-DNA methyltransferase (MGMT) has been well-characterized to be a therapeutic determinant of O(6) -alkylguanine alkylating drugs. However, the underlying mechanism and clinical relevance between MGMT and CDDP remain poorly defined in NPC. In this study, we showed that MGMT-proficient cells were highly resistant to the cytotoxic effects of CDDP as compared to MGMT-deficient cells. Further studies showed that the platinum level of DNA after CDDP exposure was significantly lower in MGMT-proficient cells than in MGMT-deficient cells. Host cell reactivation assay revealed that MGMT protected NPC cells from CDDP-induced DNA damage by enhancing DNA repair capacity. Importantly, we demonstrated for the first time that MGMT protein directly bound to CDDP-induced DNA damages. Subsequently, CDDP-bound MGMT protein became ubiquitinated and was degraded through ubiquitin-mediated proteasome system. We further analyzed the relationship between MGMT expression and clinical survivals in a cohort of 83 NPC patients. NPC patients who received CDDP-based concurrent chemoradiotherapy (CCRT), with high MGMT expression level, exhibited shorter progression-free survival (PFS; p = 0.022) and overall survival (OS; p = 0.015), than patients with low MGMT expression level. Furthermore, high MGMT expression level remained to be an independent prognostic factor for worse PFS (p = 0.01, hazard ratio 2.23) and OS (p = 0.018, hazard ratio 2.14). Our findings suggest that MGMT protein is important to determine the efficacy of CDDP in NPC.


Assuntos
Cisplatino/farmacologia , Adutos de DNA/efeitos dos fármacos , Reparo do DNA/efeitos dos fármacos , Neoplasias Nasofaríngeas/tratamento farmacológico , Neoplasias Nasofaríngeas/genética , O(6)-Metilguanina-DNA Metiltransferase/metabolismo , Compostos Organoplatínicos/farmacologia , Antineoplásicos/farmacologia , Carcinoma , Linhagem Celular Tumoral , Adutos de DNA/genética , Reparo do DNA/genética , Intervalo Livre de Doença , Humanos , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/metabolismo , Neoplasias Nasofaríngeas/patologia , Prognóstico
13.
Toxicol Appl Pharmacol ; 273(3): 492-9, 2013 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-24080332

RESUMO

Our previous studies indicated that zinc induced inflammatory response in both vascular endothelial cells and promonocytes. Here, we asked if other metals could cause the similar effect on vascular endothelial cells and tried to determine its underlying mechanism. Following screening of fifteen metals, zinc and nickel were identified with a marked proinflammatory effect, as determined by ICAM-1 and IL-8 induction, on human umbilical vein endothelial cells (HUVECs). Inhibiting protein expression of myeloid differentiation primary response protein-88 (MyD88), a Toll-like receptor (TLR) adaptor acting as a TLR-signaling transducer, significantly attenuated the zinc/nickel-induced inflammatory response, suggesting the critical roles of TLRs in the inflammatory response. Blockage of TLR-4 signaling by CLI-095, a TLR-4 inhibitor, completely inhibited the nickel-induced ICAM-1 and IL-8 expression and NFκB activation. The same CLI-095 treatment significantly blocked the zinc-induced IL-8 expression, however with no significant effect on the ICAM-1 expression and a minor inhibitory effect on the NFκB activation. The finding demonstrated the differential role of TLR-4 in regulation of the zinc/nickel-induced inflammatory response, where TLR-4 played a dominant role in NFκB activation by nickel, but not by zinc. Moreover, inhibition of NFκB by adenovirus-mediated IκBα expression and Bay 11-7025, an inhibitor of cytokine-induced IκB-α phosphorylation, significantly attenuated the zinc/nickel-induced inflammatory responses, indicating the critical of NFκB in the process. The study demonstrates the crucial role of TLRs in the zinc/nickel-induced inflammatory response in vascular endothelial cells and herein deciphers a potential important difference in NFκB activation via TLRs. The study provides a molecular basis for linkage between zinc/nickel exposure and pathogenesis of the metal-related inflammatory vascular disease.


Assuntos
Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Níquel/toxicidade , Receptor 4 Toll-Like/metabolismo , Zinco/toxicidade , Adenoviridae/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Proteínas I-kappa B/metabolismo , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , Interleucina-8/genética , Interleucina-8/metabolismo , Fator 88 de Diferenciação Mieloide/antagonistas & inibidores , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/metabolismo , Inibidor de NF-kappaB alfa , NF-kappa B/antagonistas & inibidores , NF-kappa B/genética , NF-kappa B/metabolismo , Transdução de Sinais , Sulfonamidas/farmacologia , Receptor 4 Toll-Like/antagonistas & inibidores , Receptor 4 Toll-Like/genética , Doenças Vasculares/induzido quimicamente , Doenças Vasculares/patologia
14.
Cancer Epidemiol ; 37(5): 653-9, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23752126

RESUMO

CYP2A13, a human pulmonary specific cytochrome P450 enzyme, plays an important role in susceptibility to tobacco-specific nitrosamines (TSNAs)-induced lung cancer in humans. The pattern of CYP2A13 distribution in respiratory tract affects the susceptibility of the lung to carcinogens. CYP2A13 is expressed in the epithelium of trachea and bronchi; however its pattern of expression in human lung cancer remains largely unknown. This study aimed to determine the CYP2A13 expression in specimens from human non-small cell lung carcinomas (NSCLCs), i.e., adenocarcinoma and squamous carcinoma, by immunohistochemical (IHC) analysis and to identify the potential linkage between tumor CYP2A13 levels and some clinicopathological characteristics of NSCLC patients in Taiwan. The tumor CYP2A13 IHC staining signal was strong in 76% of the 112 study subjects. Study subjects (especially non-smoking or lung adenocarcinoma patients) with higher tumor CYP2A13 levels were younger. Multiple logistic regression analysis revealed that in younger subjects (age ≤ 66) and heavy smokers (pack-years ≥ 40), the odds ratio (OR) for positive tumor CYP2A13 staining was significantly higher than that for negative tumor CYP2A13 staining. Moreover, the association of EGFR gene mutations and positive tumor CYP2A13 staining was also revealed. In conclusion, these findings suggest the potential involvement of pulmonary CYP2A13 in the early occurrence of NSCLC as well as in the development of EGFR gene mutations.


Assuntos
Hidrocarboneto de Aril Hidroxilases/biossíntese , Carcinoma Pulmonar de Células não Pequenas/enzimologia , Carcinoma Pulmonar de Células não Pequenas/genética , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/genética , Pulmão/enzimologia , Fatores Etários , Idoso , Receptores ErbB/genética , Feminino , Humanos , Imuno-Histoquímica , Masculino , Mutagênese , Mutação , Fumar/genética , Fumar/metabolismo
15.
Environ Sci Process Impacts ; 15(6): 1264-70, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23673379

RESUMO

A new and easy fast-screening test (the Ad-DR (adenoviral vector-dioxin response) bioassay) for dioxins in biological samples from highly dioxin-contaminated areas was developed. The aryl-hydrocarbon-receptor (AhR) reporter system was utilized to transport a dioxin-responsive-element (DRE) via an adenovirus vector into rat hepatoma (H4IIE) cells before each experiment; these DRE-H4IIE cells were utilized in the Ad-DR bioassay. Biological extracts were simultaneously analyzed by the Ad-DR bioassay and high resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS). A good correlation was found between the results of the HRGC/HRMS assay and those of the Ad-DR bioassay (R(2) = 0.920, p < 0.001). The bio-analytical equivalent (BEQ) value found in fish or crab caught in the abandoned pentachlorophenol plant (AP) was extremely high compared with the BEQ in fish or crab caught in two rivers nearby this abandoned plant. Dioxins were more heavily bioaccumulated in fish viscera than in fish muscles or in the whole fish. Two-way analysis of variance tests identified the significant effects of fish collection site, fish or crab tissue sample and the interaction between them on dioxin levels in the tissues of these aquatic animals. In conclusion, the Ad-DR bioassay is a useful tool to determine dioxin levels in samples of fish and crab. Compared with fish tissues, where a sample is taken (in the PCP plant or nearby rivers) is the most important factor to determine bioaccumulation of dioxins in fish.


Assuntos
Técnicas Biossensoriais/métodos , Braquiúros/metabolismo , Dioxinas/análise , Monitoramento Ambiental/métodos , Peixes/metabolismo , Poluentes Químicos da Água/análise , Animais , Engenharia Celular/métodos , Linhagem Celular Tumoral , Dioxinas/metabolismo , Vetores Genéticos/genética , Ratos , Poluentes Químicos da Água/metabolismo
16.
Respiration ; 84(4): 319-26, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22890016

RESUMO

BACKGROUND: Human CYP2A6 and CYP2A13 play important roles in metabolic activation of many pulmonary carcinogens and thus their expression and distribution may determine the pulmonary susceptibility to metabolically activated carcinogens and the following lung cancer development. Because of the 93.5% of amino acid identity between CYP2A6 and CYP2A13, generation of antibodies specific to CYP2A6 or CYP2A13 has limited immunohistochemical (IHC) analysis of CYP2A6 and CYP2A13 levels in the respiratory tract. OBJECTIVES: This study aimed to determine the differential distribution of CYP2A6 and CYP2A13 in human respiratory tissue with IHC analysis. METHODS: With computer-aided protein sequence analyses, candidate epitopes of 15 amino acids in the C-terminal domains of CYP2A6 and CYP2A13 were selected for antibody generation. Specificity of these two antibodies was confirmed with immunoblot and immunofluorescence analyses. With these two selective antibodies, the differential distribution of CYP2A6 and CYP2A13 in human respiratory tissues, including tracheae, bronchi, bronchioles and alveoli, was determined. RESULTS: IHC results showed that both CYP2A6 and CYP2A13 were markedly expressed in epithelial cells of tracheae and bronchi and that only CYP2A6 was detected in bronchiolar epithelial cells of peripheral lungs. A limitation of the present study is the cross-reactivity of our CYP2A6 antibody to the functional inactive CYP2A7. CONCLUSIONS: The differential distribution patterns of CYP2A6 and CYP2A13 in the respiratory tract are of importance in considering the pulmonary susceptibility to carcinogens and the following lung cancer development.


Assuntos
Hidrocarboneto de Aril Hidroxilases/metabolismo , Sistema Respiratório/metabolismo , Adulto , Biotransformação , Brônquios/metabolismo , Bronquíolos/metabolismo , Carcinógenos/metabolismo , Citocromo P-450 CYP2A6 , Feminino , Humanos , Imuno-Histoquímica , Neoplasias Pulmonares/etiologia , Masculino , Pessoa de Meia-Idade , Alvéolos Pulmonares/metabolismo , Distribuição Tecidual , Traqueia/metabolismo
17.
J Hazard Mater ; 196: 335-41, 2011 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-21974847

RESUMO

Our previous studies found that zinc oxide (ZnO) particles induced expression of intercellular adhesion molecule-1 (ICAM-1) protein in vascular endothelial cells via NF-κB and that zinc ions dissolved from ZnO particles might play the major role in the process. This study aimed to determine if zinc ions could cause inflammatory responses in a human promonocytic leukemia cell line HL-CZ. Conditioned media from the zinc-treated HL-CZ cells induced ICAM-1 protein expression in human umbilical vein endothelial cells (HUVEC). Zinc treatment induced chemokine and inflammatory cytokine release from HL-CZ cells. Inhibition of NFκB activity by over-expression of IκBα in HL-CZ cells did not block the conditioned medium-induced ICAM-1 protein expression in HUVEC cells. Zinc treatment induced activation of multiple immune response-related transcription factors in HL-CZ cells. These results clearly show that zinc ions induce chemokine and inflammatory cytokine release from human promonocytes, accompanied with activation of multiple immune response-related transcription factors. Our in vitro evidence in the zinc-induced inflammatory responses of vascular cells provides a critical linkage between zinc exposure and pathogenesis of those inflammatory vascular diseases.


Assuntos
Quimiocinas/metabolismo , Molécula 1 de Adesão Intercelular/biossíntese , Células Precursoras de Monócitos e Macrófagos/efeitos dos fármacos , Células Precursoras de Monócitos e Macrófagos/imunologia , Zinco/toxicidade , Adenoviridae/genética , Sequência de Bases , Técnicas de Cultura de Células , Linhagem Celular Tumoral , Quimiocinas/genética , Meios de Cultivo Condicionados , Citocinas/genética , Citocinas/metabolismo , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/imunologia , Genes Reporter , Células Endoteliais da Veia Umbilical Humana , Humanos , Luciferases de Vaga-Lume/genética , Dados de Sequência Molecular , Células Precursoras de Monócitos e Macrófagos/metabolismo , Fatores de Transcrição/genética , Acetato de Zinco/toxicidade
18.
Cancer Lett ; 309(1): 110-8, 2011 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-21684680

RESUMO

D-501036 is a promising anti-cancer compound that exhibits potent anti-proliferative activity against various types of human cancers through the induction of double strand DNA breaks. To determine drug resistance mechanism related to this class of DNA-damaging agents, a KB-derived D-501036-resistant cell line (S4) was established. Results showed that S4 cells exhibit enhanced DNA rejoining ability as compare to KB cells, through up-regulation of the non-homologous end joining activity. In conclusion, enhancement of NHEJ activity plays important role in the development of D-501036-resistance and targeting NHEJ-related molecules maybe able to overcome drug resistance to DNA damaging agents.


Assuntos
Reparo do DNA/genética , Resistencia a Medicamentos Antineoplásicos , Compostos Organosselênicos/farmacologia , Pirróis/farmacologia , Neoplasias do Colo do Útero/tratamento farmacológico , Neoplasias do Colo do Útero/genética , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Quebras de DNA de Cadeia Dupla , Resistencia a Medicamentos Antineoplásicos/genética , Feminino , Humanos , Compostos Organosselênicos/uso terapêutico , Pirróis/uso terapêutico , Regulação para Cima , Neoplasias do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/patologia
19.
Toxicol Appl Pharmacol ; 253(2): 145-52, 2011 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-21473878

RESUMO

Both cytochrome P450 2A6 (CYP2A6) and cytochrome P450 2A13 (CYP2A13) are involved in metabolic activation of tobacco-specific nitrosamines and may play important roles in cigarette smoking-induced lung cancer. Unlike CYP2A6, effects of CYP2A13 on the tobacco-specific nitrosamine-induced mutagenesis in lung cells remain unclear. This study uses a supF mutagenesis assay to examine the relative effects of CYP2A6 and CYP2A13 on metabolic activation of a tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), and its resulting mutagenesis in human lung cells. A recombinant adenovirus-mediated CYP2A6/CYP2A13 expression system was established to specifically address the relative effects of these two CYPs. Mutagenesis results revealed that both CYP2A6 and CYP2A13 significantly enhanced the NNK-induced supF mutation and that the mutagenic effect of CYP2A13 was markedly higher than that of CYP2A6. Analysis of NNK metabolism indicated that ≥70% of NNK was detoxified to 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), either with or without CYP2A6/CYP2A13 expression. Both CYP2A6 and CYP2A13 significantly enhanced the α-hydroxylation of NNK; and the α-hydroxylation activity of CYP2A13 was significantly higher than that of CYP2A6. Analysis of the NNK-related DNA adduct formation indicated that, in the presence of CYP2A13, NNK treatments caused marked increases in O(6)-methylguanine (O(6)-MeG). The present results provide the first direct in vitro evidence demonstrating the predominant roles of CYP2A13 in NNK-induced mutagenesis, possibly via metabolic activation of NNK α-hydroxylation.


Assuntos
Hidrocarboneto de Aril Hidroxilases/fisiologia , Carcinógenos/metabolismo , Nitrosaminas/metabolismo , Adenoviridae/genética , Hidrocarboneto de Aril Hidroxilases/genética , Biotransformação , Citocromo P-450 CYP2A6 , Adutos de DNA/metabolismo , Humanos , Mutagênese , Mutação
20.
J Hazard Mater ; 183(1-3): 182-8, 2010 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-20674161

RESUMO

This study investigated inflammatory effects of zinc oxide (ZnO) particles on vascular endothelial cells. The effects of 50 and 100-nm ZnO particles on human umbilical vein endothelial cells (HUVECs) were characterized by assaying cytotoxicity, cell proliferation, and glutathione levels. A marked drop in survival rate was observed when ZnO concentration was increased to 45 µg/ml. ZnO concentrations of ≤3 µg/ml resulted in increased cell proliferation, while those of ≤45 µg/ml caused dose-dependent increases in oxidized glutathione levels. Treatments with ZnO concentrations ≤45 µg/ml were performed to determine the expression of intercellular adhesion molecule-1 (ICAM-1) protein, an indicator of vascular endothelium inflammation, revealing that ZnO particles induced a dose-dependent increase in ICAM-1 expression and marked increases in NF-κB reporter activity. Overexpression of IκBα completely inhibited ZnO-induced ICAM-1 expression, suggesting NF-κB plays a pivotal role in regulation of ZnO-induced inflammation in HUVECs. Additionally, TNF-α, a typical inflammatory cytokine, induced ICAM-1 expression in an NF-κB-dependent manner, and ZnO synergistically enhanced TNF-α-induced ICAM-1 expression. Both 50 and 100-nm ZnO particles agglomerated to similar size distributions. This study reveals an important role for ZnO in modulating inflammatory responses of vascular endothelial cells via NF-κB signaling, which could have important implications for treatments of vascular disease.


Assuntos
Células Endoteliais/patologia , Inflamação/induzido quimicamente , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Óxido de Zinco/farmacologia , Morte Celular , Proliferação de Células , Células Cultivadas , Relação Dose-Resposta a Droga , Células Endoteliais/efeitos dos fármacos , Glutationa/análise , Humanos , Molécula 1 de Adesão Intercelular/análise , Nanopartículas , Transdução de Sinais/imunologia
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