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BACKGROUND: Phase angle (PhA), which is measured using bioelectrical impedance analysis, is an indicator of muscle quality and malnutrition. PhA has been shown to be correlated with sarcopenia and malnutrition; however, studies on patients with chronic obstructive pulmonary disease (COPD) are limited. In this study, we investigated the correlation between PhA and sarcopenia and malnutrition and determined the cutoff values of PhA for those in patients with COPD. METHODS: This study included 105 male patients with COPD (mean age 75.7 ± 7.7 years, mean forced expiratory volume in 1s % predicted [%FEV1] 57.0 ± 20.1%) and 12 male controls (mean age 74.1 ± 3.8 years) who were outpatients between December 2019 and March 2024. PhA was measured using the InBody S10, and its correlation with sarcopenia and malnutrition was assessed. The cutoff PhA values for sarcopenia and malnutrition were determined using receiver operating characteristic curves. RESULTS: The prevalence rates of sarcopenia and malnutrition were 31% and 22%, respectively, in patients with COPD. PhA significantly correlated with sarcopenia- and malnutrition-related indicators. Multivariate logistic regression analysis independently correlated PhA with sarcopenia and malnutrition. The cutoff values of the PhA for sarcopenia and malnutrition were 4.75° (AUC = 0.78, 95% CI = 0.68-0.88) and 4.25° (AUC = 0.75, 95% CI = 0.63-0.86), respectively. CONCLUSIONS: PhA was significantly correlated with sarcopenia and malnutrition in Japanese patients with COPD and may be a useful diagnostic indicator.
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Impedância Elétrica , Desnutrição , Doença Pulmonar Obstrutiva Crônica , Sarcopenia , Humanos , Sarcopenia/epidemiologia , Sarcopenia/etiologia , Sarcopenia/diagnóstico , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Doença Pulmonar Obstrutiva Crônica/complicações , Desnutrição/epidemiologia , Desnutrição/diagnóstico , Desnutrição/etiologia , Masculino , Idoso , Prevalência , Idoso de 80 Anos ou mais , Volume Expiratório ForçadoRESUMO
PURPOSE: Air pollutants, such as Asian sand and particulate matter (PM) 2.5, have become a global concern for causing ocular inflammation and allergic symptoms. This study, as part of an international investigation, examined the effects of eyewashes for ocular damage caused by air pollution in Indonesia. METHODS: This was a single-center, patient- and-evaluator-blinded, parallel two-arm, nonrandomized trial. In Jakarta, Indonesia, 30 eyes of 15 car commuters and 30 eyes of 15 motorcycle commuters were recruited from healthy volunteers. After commuting to work, both eyes were washed with a commercial eyewash. Before and after eyewashing, eight items of ocular surface symptoms and four items of rhinitis subjective symptoms were scored using a modified Japanese Allergic Conjunctival Disease Quality-of-Life Questionnaire. RESULTS: Five of the 12 subjective symptom scores before eyewashing were higher in motorcycle commuters than in car commuters (p < 0.05). Motorcycle commuters showed improvement in the five symptom scores of "itchy eyes, foreign body sensation, eye mucus, dryness, and eye strain" after eyewashing compared to before eyewashing (p < 0.05). In all patients, sootlike particles and ocular mucus were found in the solutions collected after eyewashing. CONCLUSION: These findings indicate that eyewashing for ocular symptoms caused by airborne particles may be effective in removing foreign particles from the ocular surface and relieving subjective symptoms.
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Poluentes Atmosféricos , Poluição do Ar , Humanos , Indonésia , Material Particulado/análise , Poluentes Atmosféricos/análise , Túnica Conjuntiva/químicaRESUMO
Skin image analysis using artificial intelligence (AI) has recently attracted significant research interest, particularly for analyzing skin images captured by mobile devices. Acne is one of the most common skin conditions with profound effects in severe cases. In this study, we developed an AI system called AcneDet for automatic acne object detection and acne severity grading using facial images captured by smartphones. AcneDet includes two models for two tasks: (1) a Faster R-CNN-based deep learning model for the detection of acne lesion objects of four types, including blackheads/whiteheads, papules/pustules, nodules/cysts, and acne scars; and (2) a LightGBM machine learning model for grading acne severity using the Investigator's Global Assessment (IGA) scale. The output of the Faster R-CNN model, i.e., the counts of each acne type, were used as input for the LightGBM model for acne severity grading. A dataset consisting of 1572 labeled facial images captured by both iOS and Android smartphones was used for training. The results show that the Faster R-CNN model achieves a mAP of 0.54 for acne object detection. The mean accuracy of acne severity grading by the LightGBM model is 0.85. With this study, we hope to contribute to the development of artificial intelligent systems to help acne patients better understand their conditions and support doctors in acne diagnosis.
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PURPOSE: To assess the effects of tranilast on degranulation and IL-13 production in mast cells activated by IL-33 and cross-linking of FcÉRI. METHODS: Bone marrow-derived mast cells (BMMCs) were sensitized with anti-DNP IgE. Sensitized cells were pretreated with tranilast and further stimulated with DNP-BSA, IL-33, or a combination of DNP-BSA and IL-33. Degranulation and the level of IL-13 release and mRNA expression in BMMCs were measured. RESULTS: Simultaneous stimulation with DNP-BSA and IL-33 resulted in marked increase in ß-hexosaminidase release. Tranilast significantly inhibited degranulation of BMMCs in the condition of combined treatment of DNP-BSA and IL-33. Combination of DNP-BSA and IL-33 induced a pronounced increase in the IL-13 release and mRNA expression. Tranilast significantly inhibited IL-13 release and mRNA expression in BMMCs stimulated by DNP-BSA and IL-33. CONCLUSIONS: Tranilast has efficacy on the inhibition of degranulation and IL-13 production in BMMCs induced by the combination of DNP-BSA and IL-33.
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Degranulação Celular/efeitos dos fármacos , Antagonistas dos Receptores Histamínicos H1/farmacologia , Interleucina-13/metabolismo , Interleucina-33/farmacologia , Mastócitos/metabolismo , Receptores de IgE/imunologia , ortoaminobenzoatos/farmacologia , Animais , Células da Medula Óssea/metabolismo , Reagentes de Ligações Cruzadas , Dinitrofenóis/farmacologia , Sinergismo Farmacológico , Interleucina-13/genética , Camundongos , Camundongos Endogâmicos C57BL , RNA Mensageiro/metabolismo , Soroalbumina Bovina/farmacologia , beta-N-Acetil-Hexosaminidases/metabolismoRESUMO
MicroRNAs (miRNAs) are small non-coding RNAs that function as endogenous silencers of target genes. Some tumor-suppressive miRNAs are known to be epigenetically silenced by promoter DNA methylation in cancer. In the present study, we aimed to identify miRNA genes that are silenced by DNA hypermethylation in hepatocellular carcinoma (HCC). We screened for miRNA genes with promoter DNA hypermethylation using a genome-wide methylation microarray analysis in HCC cells. It was found that miR-335, which is harbored within an intron of its protein-coding host gene, MEST, was downregulated by aberrant promoter hypermethylation via further methylation assays, including methylation-specific PCR, combined bisulfite and restriction analysis, bisulfite sequencing analysis and 5-aza-2'-deoxycytidine treatment. The expression levels of miR-335 significantly correlated with those of MEST, supporting the notion that the intronic miR-335 is co-expressed with its host gene. The levels of miR-335/MEST methylation were significantly higher in 18 (90%) out of 20 primary HCC tumors, compared to their non-tumor tissue counterparts (P<0.001). The expression levels of miR-335 were significantly lower in 25 (78%) out of 32 primary HCC tumors, compared to their non-tumor tissue counterparts (P=0.001). Furthermore, the expression levels of miR-335 were significantly lower in HCC tumors with distant metastasis compared to those without distant metastasis (P=0.02). In conclusion, our results indicate that expression of miR-335 is reduced by aberrant DNA methylation in HCC.
Assuntos
Carcinoma Hepatocelular/genética , Metilação de DNA , MicroRNAs/genética , Proteínas/genética , Carcinoma Hepatocelular/patologia , Epigênese Genética , Regulação Neoplásica da Expressão Gênica , Inativação Gênica , Genoma Humano , Células Hep G2 , HumanosRESUMO
BACKGROUND: The SWI/SNF chromatin remodelling complex, which contains either brahma-related gene-1 (BRG1) or brahma (BRM) as the catalytic ATPase, functions as a master regulator of gene expression. AIMS: To examine alterations of BRG1 and BRM in hepatocellular carcinoma (HCC). METHODS: We investigated DNA copy number aberrations in human HCC cell lines using a high-density oligonucleotide microarray. We determined DNA copy numbers and expression levels of BRG1 and BRM genes in primary HCC tumours, and conducted further searches for mutations in BRG1 and BRM genes. RESULTS: Homozygous deletion of the BRG1 gene was found in HCC cell line SNU398. Copy number losses of BRG1 and BRM genes were observed in 14 (26%) and 7 (13%) of 54 primary HCC tumours respectively. We found four somatic missense mutations in the BRG1 gene in two of 36 primary HCC tumours, but no mutations in BRM gene. Expression of BRM mRNA, but not BRG1 mRNA, was significantly reduced in primary HCC tumours, compared to non-tumour tissue counterparts. Immunohistochemical analyses of non-tumour liver tissues showed that BRM protein was expressed in hepatocytes and bile-duct epithelial cells, whereas BRG1 protein was expressed in bile-duct epithelial cells, but not in hepatocytes. BRM protein expression was lost in nine (22.5%) of 40 HCC tumours. Loss of BRM protein expression was significantly associated with poor overall survival. CONCLUSION: Reduced expression of BRM may contribute to the carcinogenesis of HCC. Although deletions and mutations in BRG1 gene were identified, the role of BRG1 in HCC tumourigenesis remains unclear.
Assuntos
Biomarcadores Tumorais/genética , Carcinoma Hepatocelular/genética , Proteínas Cromossômicas não Histona/genética , DNA Helicases/genética , Neoplasias Hepáticas/genética , Proteínas Nucleares/genética , Fatores de Transcrição/genética , Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/mortalidade , Linhagem Celular Tumoral , Distribuição de Qui-Quadrado , Proteínas Cromossômicas não Histona/metabolismo , Variações do Número de Cópias de DNA , DNA Helicases/metabolismo , Análise Mutacional de DNA , Feminino , Deleção de Genes , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/mortalidade , Masculino , Pessoa de Meia-Idade , Mutação de Sentido Incorreto , Proteínas Nucleares/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Polimorfismo de Nucleotídeo Único , Prognóstico , Fatores de Transcrição/metabolismoRESUMO
An increasing level of prostaglandin (PG) E(2) is involved in the progression of neuroinflammation induced by ischemia and bacterial infection. Although an imbalance in the rates of production and clearance of PGE(2) under these pathological conditions appears to affect the concentration of PGE(2) in the cerebrospinal fluid (CSF), the regulatory system remains incompletely understood. The purpose of this study was to investigate the cellular system of PGE(2) production via microsomal PGE synthetase-1 (mPGES-1), the inducible PGE(2) -generating enzyme, and PGE(2) elimination from the CSF via the blood-CSF barrier (BCSFB). Immunohistochemical analysis revealed that mPGES-1 was expressed in the soma and perivascular sheets of astrocytes, pia mater, and brain blood vessel endothelial cells, suggesting that these cells are local production sites of PGE(2) in the CSF. The in vivo PGE(2) elimination clearance from the CSF was eightfold greater than that of d-mannitol, which is considered to reflect CSF bulk flow. This process was inhibited by the simultaneous injection of unlabeled PGE(2) and ß-lactam antibiotics, such as benzylpenicillin, cefazolin, and ceftriaxone, which are substrates and/or inhibitors of organic anion transporter 3 (OAT3). The characteristics of PGE(2) uptake by the isolated choroid plexus were at least partially consistent with those of OAT3. OAT3 was able to mediate PGE(2) transport with a Michaelis-Menten constant of 4.24 µM. These findings indicate that a system regulating the PGE(2) level in the CSF involves OAT3-mediated PGE(2) uptake by choroid plexus epithelial cells, acting as a cerebral clearance pathway via the BCSFB of locally produced PGE(2) .
Assuntos
Barreira Hematoencefálica/metabolismo , Plexo Corióideo/metabolismo , Dinoprostona/metabolismo , Animais , Encéfalo/metabolismo , Immunoblotting , Imuno-Histoquímica , Oxirredutases Intramoleculares/biossíntese , Masculino , Camundongos , Transportadores de Ânions Orgânicos Sódio-Independentes/metabolismo , Prostaglandina-E Sintases , Ratos , Xenopus laevisRESUMO
Although the level of prostaglandin (PG) D(2) in cerebrospinal fluid (CSF) affects the action of D-type prostanoid receptors that promote physiological sleep, the regulatory system of PGD(2) clearance from the CSF is not fully understood. The purpose of this study was to investigate PGD(2) elimination from the CSF via the blood-CSF barrier (BCSFB). The in vivo PGD(2) elimination clearance from the CSF was 16-fold greater than that of inulin, which is considered to reflect CSF bulk flow. This process was inhibited by the simultaneous injection of unlabeled PGD(2). The characteristics of PGD(2) uptake by isolated choroid plexus were, at least partially, consistent with those of PG transporter (PGT) and organic anion transporter 3 (OAT3). Studies using an oocyte expression system showed that PGT and OAT3 were able to mediate PGD(2) transport with a Michaelis-Menten constant of 1.07 and 7.32 µM, respectively. Reverse transcription-polymerase chain reaction and immunohistochemical analyses revealed that PGT was localized on the brush-border membrane of the choroid plexus epithelial cells. These findings indicate that the system regulating the PGD(2) level in the CSF involves PGT- and OAT3-mediated PGD(2) uptake by the choroid plexus epithelial cells, acting as a pathway for PGD(2) clearance from the CSF via the BCSFB.
Assuntos
Barreira Hematoencefálica/metabolismo , Transportadores de Ânions Orgânicos Sódio-Independentes/líquido cefalorraquidiano , Transportadores de Ânions Orgânicos/líquido cefalorraquidiano , Prostaglandina D2/líquido cefalorraquidiano , Sono/fisiologia , Animais , Transporte Biológico , Plexo Corióideo/metabolismo , Cromatografia Líquida de Alta Pressão , Células Epiteliais/metabolismo , Feminino , Imuno-Histoquímica , Injeções Intraventriculares , Masculino , Taxa de Depuração Metabólica , Camundongos , Camundongos Endogâmicos , Oócitos/metabolismo , Transportadores de Ânions Orgânicos/sangue , Transportadores de Ânions Orgânicos/genética , Transportadores de Ânions Orgânicos Sódio-Independentes/sangue , Transportadores de Ânions Orgânicos Sódio-Independentes/genética , Prostaglandina D2/administração & dosagem , Prostaglandina D2/genética , Prostaglandina D2/farmacocinética , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Distribuição Tecidual , Xenopus laevisRESUMO
SOX2 is a transcription factor with a high-mobility group DNA-binding domain that functions as a master regulator during embryogenesis and organogenesis. We investigated DNA copy number aberrations in esophageal squamous cell carcinoma (ESCC) cell lines using a high-density oligonucleotide microarray and found frequent amplification at the chromosomal region 3q26. The estimated extent of the minimal overlapping region of amplification was 1.3 Mb. This chromosomal region includes a single gene, SOX2. The SOX2 protein was overexpressed in cell lines in which the gene was amplified. Knockdown experiments showed that SOX2 promotes proliferation of ESCC cells. Genes potentially modulated by SOX2 were determined by expression array analyses combined with small interfering RNA cell-transfection studies. A copy number gain of SOX2 (>2-fold) was observed in 6 of the 40 primary ESCCs (15%). Immunohistochemical study revealed that expression of the SOX2 protein was significantly elevated in 62 of the 89 ESCC tumors (70%), compared with their nontumorous counterparts, and that upregulated expression of SOX2 was associated with poor differentiation of ESCC. Our results suggest that SOX2 is likely to be a target of the 3q26 amplification and may therefore be involved in the development or progression of ESCC.
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Carcinoma de Células Escamosas/genética , Cromossomos Humanos Par 3/genética , Neoplasias Esofágicas/genética , Amplificação de Genes , Fatores de Transcrição SOXB1/genética , Carcinoma de Células Escamosas/patologia , Diferenciação Celular/genética , Linhagem Celular Tumoral , Proliferação de Células , Variações do Número de Cópias de DNA/genética , Regulação para Baixo/genética , Neoplasias Esofágicas/patologia , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Genoma Humano/genética , Humanos , Polimorfismo de Nucleotídeo Único/genética , RNA Interferente Pequeno/metabolismo , Fatores de Transcrição SOXB1/metabolismo , Transativadores/genética , Transativadores/metabolismo , Fatores de Transcrição , Transfecção , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismoRESUMO
DNA copy number aberrations in human hepatocellular carcinoma (HCC) cell lines were investigated using a high-density oligonucleotide microarray, and a novel amplification at the chromosomal region 7q21 was detected. Molecular definition of the amplicon indicated that PEG10 (paternally expressed gene 10), a paternally expressed imprinted gene, was amplified together with CDK14 (cyclin-dependent kinase 14; previously PFTAIRE protein kinase 1, PFTK1) and CDK6 (cyclin-dependent kinase 6). An increase in PEG10 copy number was detected in 14 of 34 primary HCC tumors (41%). PEG10, but not CDK14 or CDK6, was significantly overexpressed in 30 of 41 tumors (73%) from HCC patients, compared with their nontumorous counterparts. These results suggest that PEG10 is a probable target, acting as a driving force for amplification of the 7q21 region, and may therefore be involved in the development or progression of HCCs.
Assuntos
Carcinoma Hepatocelular/genética , Cromossomos Humanos Par 7 , Amplificação de Genes , Neoplasias Hepáticas/genética , Proteínas/genética , Adulto , Idoso , Proteínas Reguladoras de Apoptose , Proteína Quinase CDC2/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Cromossomos Humanos Par 7/genética , Quinase 6 Dependente de Ciclina/genética , Quinases Ciclina-Dependentes/genética , Proteínas de Ligação a DNA , Progressão da Doença , Feminino , Amplificação de Genes/fisiologia , Dosagem de Genes , Regulação Neoplásica da Expressão Gênica , Humanos , Hibridização in Situ Fluorescente , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Proteínas de Ligação a RNA , Regulação para CimaRESUMO
In clinical practice, photophobia resulting from persistent mydriasis may be associated with dysfunction of ocular parasympathetic nerves or primary iris lesions. We encountered a 5-year-old Miniature Dachshund and a 7-year-old Shih Tzu with mydriasis, abnormal pupillary light reflexes, and photophobia. Except for sustained mydriasis and photophobia, no abnormalities were detected on general physical examination or ocular examination of either dog. We performed pharmacological examinations using 0.1% and 2% pilocarpine to evaluate and diagnose parasympathetic denervation of the affected pupillary sphincter muscles. On the basis of the results, we diagnosed a pupillary abnormality due to parasympathetic dysfunction and not to overt primary iris lesions. The test revealed that neuroanatomic localization of the lesion was postciliary ganglionic in the first dog.
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Doenças do Sistema Nervoso Autônomo/veterinária , Doenças do Cão/fisiopatologia , Midríase/veterinária , Animais , Anti-Inflamatórios/uso terapêutico , Doenças do Sistema Nervoso Autônomo/complicações , Doenças do Sistema Nervoso Autônomo/tratamento farmacológico , Doenças do Sistema Nervoso Autônomo/fisiopatologia , Doenças do Cão/tratamento farmacológico , Cães , Feminino , Inflamação/tratamento farmacológico , Inflamação/prevenção & controle , Inflamação/veterinária , Masculino , Meiose , Mióticos/uso terapêutico , Midríase/tratamento farmacológico , Midríase/etiologia , Midríase/fisiopatologia , Fotofobia/etiologia , Fotofobia/veterinária , Pilocarpina/uso terapêutico , Prednisolona/uso terapêuticoRESUMO
PURPOSE: Steatosis is a histological finding associated with the progression of chronic hepatitis C. The aims of this study were to elucidate risk factors associated with steatosis and to evaluate the association between steatosis and hepatic expression of genes regulating lipid metabolism. METHODS: We analyzed 297 Japanese patients infected with hepatitis C virus and a subgroup of 100 patients who lack metabolic factors for steatosis. We determined intrahepatic mRNA levels of 18 genes regulating lipid metabolism in these 100 patients using real-time reverse transcription-polymerase chain reaction. Levels of peroxisome proliferator-activated receptor alpha and sterol regulatory element-binding protein 1 proteins were assessed by immunohistochemistry. RESULTS: Steatosis was present in 171 (57%) of 297 patients. The presence of steatosis was independently associated with a higher body mass index, higher levels of gamma-glutamyl transpeptidase and triglyceride, and a higher fibrosis stage. Steatosis was present in 43 (43%) of 100 patients lacking metabolic factors. Levels of mRNA and protein of peroxisome proliferator-activated receptor alpha, which regulates beta-oxidation of fatty acid, were lower in patients with steatosis than in patients without steatosis. CONCLUSIONS: These findings indicate that impaired degradation of lipid may contribute to the development of hepatitis C virus-related steatosis.
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Fígado Gorduroso/etiologia , Regulação da Expressão Gênica , Hepatite C/complicações , Metabolismo dos Lipídeos/genética , Adulto , Idoso , Fígado Gorduroso/epidemiologia , Fígado Gorduroso/genética , Feminino , Hepatite C/genética , Humanos , Japão/epidemiologia , Fígado/metabolismo , Fígado/fisiopatologia , Fígado/virologia , Masculino , Pessoa de Meia-Idade , PPAR alfa/metabolismo , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Risco , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismo , Adulto JovemRESUMO
Sphingosine 1-phosphate (S1P) and lysophosphatidic acid (LPA) are simple bioactive lysophospholipids which exhibit an effect on blood vessels via their G protein-coupled receptors. The purpose of this study was (i) to clarify the impact of S1P and LPA on the release of taurine as an organic osmolyte from rat retinal vascular endothelial cells (RVECs) under hypoosmotic stress and (ii) to quantify the gene expression levels of S1P and LPA receptors in RVECs. When cultured RVECs (TR-iBRB2 cells) that had been preloaded with [(3)H]taurine were exposed to hypotonic buffer (230 mOsm) for 1 to 10 min, [(3)H]taurine release from the cells was several times greater than that using an isotonic buffer (300 mOsm). S1P and LPA significantly enhanced the [(3)H]taurine release under hypotonic conditions in a time- and concentration-dependent manner, whereas S1P and LPA had no significant effect under isotonic conditions. Quantitative real-time PCR revealed that freshly isolated RVECs predominantly express mRNAs for S1P(1), S1P(4) and LPA(4). The S1P-enhanced [(3)H]taurine release under hypoosmotic conditions was significantly inhibited by an S1P(1) receptor antagonist. Inhibitor of the small GTPase Rho, C3 exotoxin, attenuated S1P- and LPA-enhanced [(3)H]taurine release. These results suggest that S1P and LPA play a novel role in the regulation of osmolyte efflux from RVECs in response to hypoosmotic stress via the activation of their specific receptors.
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Células Endoteliais/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Lisofosfolipídeos/farmacologia , Vasos Retinianos/efeitos dos fármacos , Esfingosina/análogos & derivados , Taurina/metabolismo , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Células Endoteliais/metabolismo , Endotélio Vascular/metabolismo , Expressão Gênica , Lisofosfolipídeos/genética , Lisofosfolipídeos/metabolismo , Masculino , Pressão Osmótica , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptores de Ácidos Lisofosfatídicos/genética , Receptores de Ácidos Lisofosfatídicos/metabolismo , Receptores de Lisoesfingolipídeo/genética , Receptores de Lisoesfingolipídeo/metabolismo , Vasos Retinianos/metabolismo , Esfingosina/genética , Esfingosina/metabolismo , Esfingosina/farmacologiaRESUMO
AIM: To determine whether JUN (the oncogene encoding c-Jun protein) is amplified and overexpressed in hepatocellular carcinoma (HCC). MATERIALS AND METHODS: DNA copy number aberrations were investigated using a high-density oligonucleotide microarray. DNA copy numbers were determined by fluorescence in situ hybridization. Genomic DNA and mRNA were quantified using real-time quantitative PCR. RESULTS: A novel amplification was found at the chromosomal region 1p32-31 in a JHH-2 HCC cell line within which JUN is amplified and overexpressed. However, no copy number gain of JUN (>2-fold) was observed in 34 primary HCC tumors. Rather, a loss of JUN (<0.5-fold) was seen in 13 (38%) out of the 34 tumors and expression of JUN was significantly lower in 26 (70%) out of the 37 HCC tumors compared with their nontumorous counterparts. CONCLUSION: Although JUN was amplified and overexpressed in JHH-2 HCC cells, amplification and overexpression of JUN may be rare in primary HCCs.
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Carcinoma Hepatocelular/genética , Aberrações Cromossômicas , Amplificação de Genes , Genes jun , Neoplasias Hepáticas/genética , Polimorfismo de Nucleotídeo Único/genética , Carcinoma Hepatocelular/patologia , Cromossomos Humanos Par 1/genética , Dosagem de Genes , Humanos , Hibridização in Situ Fluorescente , Neoplasias Hepáticas/patologia , Hibridização de Ácido Nucleico , Análise de Sequência com Séries de Oligonucleotídeos , Células Tumorais CultivadasRESUMO
AIMS: Hepatic steatosis and iron cause oxidative stress, thereby progressing steatosis to steatohepatitis. We quantified the expression of genes involved in the metabolism of fatty acids and iron in patients with nonalcoholic fatty liver disease (NAFLD). METHODS: The levels of transcripts for the following genes were quantified from biopsy specimens of 74 patients with NAFLD: thioredoxin (Trx), fatty acid transport protein 5 (FATP5), sterol regulatory element-binding protein 1c (SREBP1c), fatty acid synthase (FASN), acetyl-coenzyme A carboxylase (ACAC), peroxisome proliferative activated receptor alpha (PPARalpha), cytochrome P-450 2E1 (CYP2E1), acyl-coenzyme A dehydrogenase (ACADM), acyl-coenzyme A oxidase (ACOX), microsomal triglyceride transfer protein (MTP), transferrin receptor 1 (TfR1), transferrin receptor 2 (TfR2) and hepcidin. Twelve samples of human liver RNA were used as controls. Histological evaluation followed the methods of Brunt. RESULTS: The levels of all genes were significantly higher in the NAFLD patients than in controls. The Trx level increased as the stage progressed. The levels of FATP5, SREBP1c, ACAC, PPARalpha, CYP2E1, ACADM and MTP significantly decreased as the stage and grade progressed (P < 0.05). Hepatic iron score (HIS) increased as the stage progressed. The TfR1 level significantly increased as the stage progressed (P < 0.05), whereas TfR2 level significantly decreased (P < 0.05). The ratio of hepcidin mRNA/ferritin (P < 0.001) or hepcidin mRNA/HIS (P < 0.01) was significantly lower in NASH patients than simple steatosis patients. CONCLUSIONS: Steatosis-related metabolism is attenuated as NAFLD progresses, whereas iron-related metabolism is exacerbated. Appropriate therapies should be considered on the basis of metabolic changes.
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RhoA, a member of the Rho family of small GTPases, directs the organization of the actin cytoskeleton and is involved in regulating cell shape and movement. Its activity is negatively regulated by p190-B RhoGAP (GTPase-activating protein). We investigated DNA copy number aberrations in human hepatocellular carcinoma and esophageal squamous cell carcinoma cell lines using a high-density oligonucleotide microarray and found a novel amplification at chromosomal region 14q12. We identified ARHGAP5 (the gene encoding p190-B RhoGAP) as a probable target for the amplification at 14q12, and our results showed that p190-B RhoGAP promotes cells spreading and migration by negatively regulating RhoA activity in Huh-7 hepatocellular carcinoma cells.
Assuntos
Carcinoma Hepatocelular/metabolismo , Proteínas Ativadoras de GTPase/metabolismo , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Cromossomos Humanos Par 14/ultraestrutura , Citoesqueleto/metabolismo , DNA/metabolismo , Progressão da Doença , Humanos , Hibridização in Situ Fluorescente , Interferência de RNA , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
High-density single nucleotide polymorphism (SNP) array analysis revealed novel amplification at 1q21 in cell lines derived from hepatocellular carcinomas (HCCs). Fluorescence in situ hybridization and real-time quantitative polymerase chain reaction studies verified amplification at 1q21. An increase in copy number at the region was detected in 32 of the 36 primary HCC tumors (89%). To identify the targets for amplification, we examined 19 HCC cell lines for expression levels of all 26 genes located within the 700-kb amplified region. Five genes were overexpressed in cell lines with amplification at 1q21. Among these, CREB3L4 (cAMP responsive element binding protein 3-like 4), INTS3 (integrator complex subunit 3), and SNAPAP (SNAP-associated protein) were significantly overexpressed in tumors from 18 HCC patients, compared with counterpart nontumorous tissues. The findings suggest that CREB3L4, INTS3, and SNAPAP are probable targets for the amplification mechanism and may therefore be involved, together or separately, in the development or progression of HCCs.