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1.
J Oncol ; 2022: 9188920, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36245989

RESUMO

YTH domain-containing 2 (YTHDC2) is known to be an important regulator for RNA metabolism. Here, we show that YTHDC2 is essential for breast cancer tumorigenesis and metastasis. We examined YTHDC2 expression levels by immunohistochemistry in human breast tumor tissues from 99 patients and found a significantly positive correlation between the YTHDC2 expression level and the tumor stage. We established YTHDC2-knocked-down cell lines using four breast cancer cell lines with different subtypes. Knockdown of YTHDC2 attenuated the sphere-forming and the metastatic ability of breast cancer cells. Although stemness and EMT markers, such as SOX2, c-MYC, and NANOG, were downregulated in several YTHDC2-knocked-down breast cancer cells, a common target gene of YTHDC2 in breast cancer cells was not identified. These findings suggest that while YTHDC2 is involved in malignant progression of breast cancers, the mechanism by which YTHDC2 regulates those phenotypes is different between subtypes of breast cancers.

2.
Biosci Biotechnol Biochem ; 85(1): 85-91, 2021 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-33577659

RESUMO

Sulfoglycolipid, SQAP, is a radiosensitizing agent that makes tumor cells more sensitive to radiation therapy. A previous study revealed that SQAP induced the degradation of hypoxia-inducible factor-1α (HIF-1α) and inhibited angiogenesis in a hepatoma model mouse. Herein, we examined the biological activities of SQAP against hepatocarcinoma cells under low oxygen conditions. Cell growth inhibition of SQAP under hypoxic conditions was significantly higher than that under normoxic conditions. In addition, SQAP was found to impair the expression of histone deacetylase (HDAC) under low oxygen conditions. Our present data suggested that SQAP induced the degradation of HIF-1α and then decreased the expression of HDAC1. Unlike known HDAC inhibitors, SQAP increased the acetylation level of histone in cells without inhibition of enzymatic activity of HDACs. Our data demonstrated hypoxia-specific unique properties of SQAP.


Assuntos
Morte Celular/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glicolipídeos/química , Glicolipídeos/farmacologia , Histona Desacetilase 1/metabolismo , Hipóxia Tumoral/efeitos dos fármacos , Acetilação/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Histonas/metabolismo , Humanos
3.
Vet Med Sci ; 7(2): 577-585, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33210459

RESUMO

The cell surface glycoprotein CD44 has various types of splicing variants, which contribute to its multiple distinct cellular functions. Recently, it was reported that the CD44v8-10 isoform interacts with the system Xc(-) transporter-related protein (xCT), and inhibits the accumulation of reactive oxygen species by promoting the synthesis of the antioxidant glutathione in human tumour cells. In this study, we investigated the expression and function of CD44 variants and xCT in canine tumours. From semi-quantitative reverse transcription polymerase chain reaction analysis, the mRNA expression of the CD44v8-10 isoform was observed in canine tumour tissues as well as human cases. The overexpression of CD44v8-10 may promote the synthesis of glutathione and enhance the resistance to radiation of canine breast tumour cells. Furthermore, canine xCT mRNA expression was significantly upregulated in the canine breast tumour tissues as compared to the normal tissues surrounding the tumours. To investigate the function of canine xCT, we treated canine tumour cells with the xCT inhibitor sulfasalazine. Consequently, the sulfasalazine-treated cells were more sensitive to oxidative stress than the non-treated cells. Taken together, these results suggested that CD44v8-10 and xCT play important roles in the therapy resistance of canine tumours as well as human tumours.


Assuntos
Sistemas de Transporte de Aminoácidos Acídicos/genética , Doenças do Cão/genética , Regulação Neoplásica da Expressão Gênica , Receptores de Hialuronatos/genética , Sistemas de Transporte de Aminoácidos Acídicos/antagonistas & inibidores , Sistemas de Transporte de Aminoácidos Acídicos/metabolismo , Animais , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama , Doenças do Cão/metabolismo , Cães , Feminino , Glutationa/metabolismo , Receptores de Hialuronatos/metabolismo , Isoformas de Proteínas , Espécies Reativas de Oxigênio/metabolismo , Sulfassalazina/farmacologia , Regulação para Cima
4.
Cancers (Basel) ; 12(10)2020 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-32998263

RESUMO

Numerous findings have indicated that CSCs, which are present at a low frequency inside primary tumors, are the main cause of therapy resistance and cancer recurrence. Although various therapeutic methods targeting CSCs have been attempted for eliminating cancer cells completely, the complicated characteristics of CSCs have hampered such attempts. In analyzing the biological properties of CSCs, it was revealed that CSCs have a peculiar metabolism that is distinct from non-CSCs to maintain their stemness properties. The CSC metabolism involves not only the catabolic and anabolic pathways, but also intracellular signaling, gene expression, and redox balance. In addition, CSCs can reprogram their metabolism to flexibly respond to environmental changes. In this review, we focus on the flexible metabolic mechanisms of CSCs, and highlight the new therapeutics that target CSC metabolism.

5.
Bioorg Med Chem ; 27(23): 115149, 2019 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-31679979

RESUMO

Pyrenocine A, a phytotoxin, was found to exhibit cytotoxicity against cancer cells with an IC50 value of 2.6-12.9 µM. Live cell imaging analysis revealed that pyrenocine A arrested HeLa cells at the M phase with characteristic ring-shaped chromosomes. Furthermore, as a result of immunofluorescence staining analysis, we found that pyrenocine A resulted in the formation of monopolar spindles in HeLa cells. Monopolar spindles are known to be induced by inhibitors of the kinesin motor protein Eg5 such as monastrol and STLC. Monastrol and STLC induce monopolar spindle formation and M phase arrest via inhibition of the ATPase activity of Eg5. Interestingly, our data revealed that pyrenocine A had no effect on the ATPase activity of Eg5 in vitro, which suggested the compound induces a monopolar spindle by an unknown mechanism. Structure-activity relationship analysis indicates that the enone structure of pyrenocine A is likely to be important for its cytotoxicity. An alkyne-tagged analog of pyrenocine A was synthesized and suppressed proliferation of HeLa cells with an IC50 value of 2.3 µM. We concluded that pyrenocine A induced monopolar spindle formation by a novel mechanism other than direct inhibition of Eg5 motor activity, and the activity of pyrenocine A may suggest a new anticancer mechanism.


Assuntos
Antineoplásicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Fuso Acromático/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Células HeLa , Humanos , Neoplasias/tratamento farmacológico , Pirimidinas/farmacologia , Pironas/farmacologia , Tionas/farmacologia
6.
Xenobiotica ; 49(3): 346-362, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29543539

RESUMO

Sulfoquinovosylacylpropanediol (SQAP) is a novel potent radiosensitizer that inhibits angiogenesis in vivo and results in increased oxigenation and reduced tumor volume. We investigated the distribution, metabolism, and excretion of SQAP in male KSN-nude mice transplanted with a human pulmonary carcinoma, Lu65. For the metabolism analysis, a 2 mg (2.98 MBq)/kg of [glucose-U-14C]-SQAP (CP-3839) was intravenously injected. The injected SQAP was decomposed into a stearic acid and a sulfoquinovosylpropanediol (SQP) in the body. The degradation was relatively slow in the carcinoma tissue.1,3-propanediol[1-14C]-SQAP (CP-3635) was administered through intravenous injection of a 1 mg (3.48 MBq)/kg dose followed by whole body autoradiography of the mice. The autoradiography analysis demonstrated that SQAP rapidly distributed throughout the whole body and then quickly decreased within 4 hours except the tumor and excretion organs such as liver, kidney. Retention of SQAP was longer in tumor parts than in other tissues, as indicated by higher levels of radioactivity at 4 hours. The radioactivity around the tumor had also completely disappeared within 72 hours.


Assuntos
Glicolipídeos/farmacocinética , Radiossensibilizantes/farmacocinética , Administração Intravenosa , Animais , Autorradiografia , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Glicolipídeos/administração & dosagem , Glicolipídeos/uso terapêutico , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Camundongos Nus , Radiossensibilizantes/administração & dosagem , Radiossensibilizantes/uso terapêutico , Espectrometria de Massas em Tandem
7.
Vet Med Sci ; 5(1): 19-29, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30265453

RESUMO

The balance between pro- and anti-angiogenic signalling is tightly regulated in normal tissues to maintain the functions of the vasculature. In contrast, the overproduction of angiogenic factors and enhanced angiogenesis are frequently observed in several types of tumours. Although there have been many reports on the correlation between tumour progression and angiogenesis in humans, little is known about tumour angiogenesis in canines. Hence, we attempted to clarify whether angiogenesis contributes to tumour progression in canines as well as humans. In this study, we investigated the expression of several angiogenesis-related genes, including CD34, VEGF-A, VEGFR-1, VEGFR-2, Ang-1, Ang-2, Tie1, and Tie2, in 66 canine tumour tissues and in the normal tissues surrounding the tumours by quantitative real-time PCR analysis. Our comparative analysis between canine tumour tissues and normal tissues revealed that several angiogenesis-related genes, such as vascular endothelial growth factor (VEGF) and VEGF-receptor genes, were significantly upregulated in canine tumour tissues when compared to the normal tissues. We also found that the angiopoietin (Ang)-1/Ang-2 gene expression ratio was lower in canine tumour tissues than in the normal tissues, suggesting less association between vascular endothelial cells and perivascular cells in the canine tumour tissues. Taken together, our results suggest that several angiogenesis-related genes may contribute to the malignant progression of canine tumours via tumour angiogenesis.


Assuntos
Doenças do Cão/metabolismo , Regulação Neoplásica da Expressão Gênica , Neoplasias/veterinária , Neovascularização Patológica/veterinária , Transcriptoma , Animais , Doenças do Cão/genética , Cães , Neoplasias/genética , Neoplasias/metabolismo , Neovascularização Patológica/genética , Neovascularização Patológica/metabolismo , RNA/genética , RNA/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Regulação para Cima
8.
Cancer Lett ; 376(1): 34-42, 2016 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-26996300

RESUMO

YTH domain containing 2 (YTHDC2) is a member of the DExD/H-box family of ATP-dependent RNA helicases. We previously found that YTHDC2 expression is up-regulated in several human cancer cells. In this study, we demonstrate novel roles for YTHDC2 in metastasis of colon tumor cells via translation-dependent pathway. Knockdown of YTHDC2 attenuated protein expression of metastasis-related genes, such as hypoxia-inducible factor-1alpha (HIF-1α), and inhibited metastasis of colon tumor cells in vitro and in vivo. To confirm that YTHDC2 promotes translation initiation by unwinding the 5'-untranslated region (5'UTR) of mRNA, we constructed a firefly luciferase reporter containing the 5'UTR of the HIF-1α mRNA and showed reduction in luciferase activity in YTHDC2-silenced cells. Furthermore, we examined expression levels of YTHDC2 by immunohistochemical staining in human colon cancer tissues from 72 patients and found a significantly positive correlation between YTHDC2 expression and the tumor stage, including metastasis. In conclusion, these results suggest that the RNA helicase YTHDC2 contributes to colon tumor metastasis by promoting translation of HIF-1α and that YTHDC2 is potentially a diagnostic marker and target gene for treating colon cancer patients.


Assuntos
Adenocarcinoma/enzimologia , Adenosina Trifosfatases/metabolismo , Movimento Celular , Neoplasias do Colo/enzimologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/biossíntese , RNA Mensageiro/metabolismo , Regiões 5' não Traduzidas , Adenocarcinoma/genética , Adenocarcinoma/secundário , Adenosina Trifosfatases/genética , Animais , Células COS , Chlorocebus aethiops , Neoplasias do Colo/genética , Neoplasias do Colo/patologia , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Genes Reporter , Células HCT116 , Células HT29 , Xenoenxertos , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Camundongos Endogâmicos BALB C , Camundongos Nus , Metástase Neoplásica , Estadiamento de Neoplasias , Transplante de Neoplasias , Proteínas Nucleares/metabolismo , RNA Helicases , Interferência de RNA , RNA Mensageiro/genética , Transdução de Sinais , Fatores de Tempo , Transfecção , Hipóxia Tumoral , Proteína 1 Relacionada a Twist/metabolismo , Regulação para Cima
9.
Sci Rep ; 5: 15136, 2015 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-26456697

RESUMO

SQAP is a novel and promising anticancer agent that was obtained by structural modifications from a natural compound. SQAP inhibits angiogenesis in vivo resulting in increased hypoxia and reduced tumor volume. In this study, the mechanism by which SQAP modifies the tumor microenvironment was revealed through the application of a T7 phage display screening. This approach identified five SQAP-binding proteins including sterol carrier protein 2, multifunctional enzyme type 2, proteasomal ubiquitin receptor, UV excision repair protein and focal adhesion kinase (FAK). All the interactions were confirmed by surface plasmon resonance analysis. Since FAK plays an important role in cell turnover and angiogenesis, the influence of SQAP on FAK was the principal goal of this study. SQAP decreased FAK phosphorylation and cell migration in human umbilical vein endothelial cells and A549 cancer cells. These findings suggest that inhibition of FAK phosphorylation works as the mechanism for the anti-angiogenesis activity of SQAP.


Assuntos
Antineoplásicos/farmacologia , Carcinoma de Células Pequenas/tratamento farmacológico , Quinase 1 de Adesão Focal/antagonistas & inibidores , Glicolipídeos/farmacologia , Neoplasias Pulmonares/tratamento farmacológico , Sequência de Aminoácidos , Animais , Antineoplásicos/síntese química , Sítios de Ligação , Carcinoma de Células Pequenas/enzimologia , Carcinoma de Células Pequenas/genética , Carcinoma de Células Pequenas/patologia , Proteínas de Transporte/química , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Enzimas Reparadoras do DNA/química , Enzimas Reparadoras do DNA/genética , Enzimas Reparadoras do DNA/metabolismo , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Quinase 1 de Adesão Focal/química , Quinase 1 de Adesão Focal/genética , Quinase 1 de Adesão Focal/metabolismo , Glicolipídeos/síntese química , Células Endoteliais da Veia Umbilical Humana , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Glicoproteínas de Membrana/química , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Camundongos , Camundongos Nus , Simulação de Acoplamento Molecular , Dados de Sequência Molecular , Biblioteca de Peptídeos , Proteína Multifuncional do Peroxissomo-2/química , Proteína Multifuncional do Peroxissomo-2/genética , Proteína Multifuncional do Peroxissomo-2/metabolismo , Fosforilação/efeitos dos fármacos , Ligação Proteica , Ensaios Antitumorais Modelo de Xenoenxerto
10.
Pharmacology ; 96(5-6): 210-6, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26329263

RESUMO

Cilostazol and L-carnitine have been used as a first-line drug and supplement, respectively, in patients with peripheral arterial disease with intermittent claudication. In this study, the effect of the combination of cilostazol and L-carnitine has been investigated in rats with unilateral hindlimb ischemia. For 28 days, cilostazol and L-carnitine were administrated separately or as a combination. The distance walked before gait disturbance developed was measured using a treadmill for 5 days a week. The capillary density of the ischemic hindlimb was evaluated by immunohistochemical staining at days 7, 14, 21, and 28. Angiogenic gene expression was measured by real-time RT-PCR at days 7 and 28. The greatest increase in the distance was observed in the combination therapy group when compared to the other groups. The capillary density in the adductor muscles of rats treated with cilostazol alone and combination therapy increased at day 28. Angiopoietin-2/Angiopoietin-1 expression ratios were higher, suggesting the promotion of angiogenesis, with cilostazol alone and combination therapy at day 7. This is the first study to show functional improvement of the hind limb following combination therapy with cilostazol and L-carnitine in experimental animals. This study also revealed that cilostazol promotes angiogenesis, and L-carnitine additively contributes to functional improvement via a non-angiogenic mechanism.


Assuntos
Carnitina/uso terapêutico , Doença Arterial Periférica/tratamento farmacológico , Tetrazóis/uso terapêutico , Vasodilatação/efeitos dos fármacos , Caminhada , Proteínas Angiogênicas/genética , Proteínas Angiogênicas/metabolismo , Animais , Carnitina/administração & dosagem , Cilostazol , Modelos Animais de Doenças , Quimioterapia Combinada , Teste de Esforço/efeitos dos fármacos , Membro Posterior/irrigação sanguínea , Isquemia/tratamento farmacológico , Isquemia/fisiopatologia , Masculino , Músculo Esquelético/irrigação sanguínea , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Doença Arterial Periférica/fisiopatologia , Ratos Sprague-Dawley , Tetrazóis/administração & dosagem
11.
Eur J Immunol ; 44(11): 3220-31, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25196058

RESUMO

While the presentation mechanism of antigenic peptides derived from exogenous proteins by MHC class II molecules is well understood, relatively little is known about the presentation mechanism of endogenous MHC class II-restricted antigens. We therefore screened a chemical library of 200 compounds derived from natural products to identify inhibitors of the presentation of endogenous MHC class II-restricted antigens. We found that pyrenocine B, a compound derived from the fungus Pyrenochaeta terrestris, inhibits presentation of endogenous MHC class II-restricted minor histocompatibility antigen IL-4 inducible gene 1 (IL4I1) by primary dendritic cells (DCs). Phage display screening and surface plasmon resonance (SPR) analysis were used to investigate the mechanism of suppressive action by pyrenocine B. EpsinR, a target molecule for pyrenocine B, mediates endosomal trafficking through binding of soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs). Lentiviral-mediated short hairpin (sh) RNA downregulation of EpsinR expression in DCs resulted in a decrease in the responsiveness of CD4+ T cells. Our data thus suggest that EpsinR plays a role in antigen presentation, which provides insight into the mechanism of presentation pathway of endogenous MHC class II-restricted antigen.


Assuntos
Proteínas Adaptadoras de Transporte Vesicular/imunologia , Apresentação de Antígeno/efeitos dos fármacos , Antígenos de Histocompatibilidade Classe II/imunologia , Proteínas Adaptadoras de Transporte Vesicular/antagonistas & inibidores , Proteínas Adaptadoras de Transporte Vesicular/genética , Animais , Apresentação de Antígeno/imunologia , Linfócitos T CD4-Positivos/imunologia , Técnicas de Visualização da Superfície Celular , Células Dendríticas/imunologia , Flavoproteínas/antagonistas & inibidores , Flavoproteínas/biossíntese , Proteínas Fúngicas/farmacologia , L-Aminoácido Oxidase , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Pironas/farmacologia , Interferência de RNA , RNA Interferente Pequeno , Proteínas SNARE/imunologia , Ressonância de Plasmônio de Superfície
12.
Southeast Asian J Trop Med Public Health ; 45(2): 395-401, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24968681

RESUMO

A lethargic household dog was referred to a private hospital in Japan. Diagnosis was carried out by the polymerase chain reaction (PCR) method developed for human Orientia tsutsugamushi infection using the dog's anticoagulated peripheral blood. Karp, Kato and Kuroki-type genomes were detected and the dog was diagnosed with O. tsutsugamushi infection. These findings demonstrate that dogs can act as a host for O. tsutsugamushi and the PCR method developed for human beings can be used for the diagnosis of canine O. tsutsugamushi infection. A concurrent epidemiological study examined 10 asymptomatic dogs that were fed in the same area as the sick dog. Kuroki-type genome in all dogs, Gilliam-type genome in 6 dogs and Kawasaki-type genome in 3 dogs were detected. These results provide further evidence that dogs can be naturally infected with O. tsutsugamushi outdoors and that dogs play a role as a host in the lifecycle of O. tsutsugamushi.


Assuntos
Doenças do Cão/diagnóstico , Doenças do Cão/epidemiologia , Orientia tsutsugamushi/isolamento & purificação , Tifo por Ácaros/diagnóstico , Tifo por Ácaros/epidemiologia , Tifo por Ácaros/veterinária , Animais , Doenças do Cão/genética , Cães , Genoma Bacteriano , Japão/epidemiologia , Orientia tsutsugamushi/genética , Reação em Cadeia da Polimerase , Tifo por Ácaros/genética , Tifo por Ácaros/microbiologia
13.
Gene ; 535(1): 24-32, 2014 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-24269672

RESUMO

We previously demonstrated that a cellular factor, cyclosporin A (CsA) associated helicase-like protein (CAHL) that is identical to YTH domain containing 2 (YTHDC2), forms trimer complex with cyclophilin B and NS5B of hepatitis C virus (HCV) and facilitates HCV genome replication. Gene expression of YTHDC2 was shown in tumor cell lines and tumor necrosis factor (TNF)-α-treated hepatocytes, but not in untreated. However, the function of YTHDC2 in the tumor cells and the mechanism by which the YTHDC2 gene is transcribed in these cells is largely unknown. We first evaluated that the role of YTHDC2 in the proliferation of hepatocellular carcinoma (HCC) cell line Huh7 using RNA interference and found that YTHDC2-downregulated Huh7 were significantly decreased cell growth as compared to control. We next demonstrated that the cAMP response element (CRE) site in the promoter region of the YTHDC2 gene is critical for YTHDC2 transcription. To further investigate the transcription factors bound to the CRE site, we performed chromatin immunoprecipitation assays. Our findings demonstrate that c-Jun and ATF-2 bind to the CRE site in Huh7, and that TNF-α induces the biological activity of these transcription factors in hepatocytes as well as Huh7. Moreover, treatment with the HDAC inhibitor, trichostatin A (TSA), reduces YTHDC2 expression in Huh7 and in TNF-α-stimulated hepatocytes. Collectively, these data show that YTHDC2 plays an important role in tumor cells growth and activation/recruitment of c-Jun and ATF-2 to the YTHDC2 promoter is necessary for the transcription of YTHDC2, and that HDAC activity is required for the efficient expression of YTHDC2 in both of hepatocyte and HCC cells.


Assuntos
Proteínas de Ligação a RNA/genética , Transcrição Gênica , Fator de Necrose Tumoral alfa/metabolismo , Sequência de Bases , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Proliferação de Células , Ilhas de CpG , Metilação de DNA , Primers do DNA , Técnicas de Silenciamento de Genes , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Regiões Promotoras Genéticas , Interferência de RNA , Reação em Cadeia da Polimerase Via Transcriptase Reversa
14.
Microbiol Immunol ; 56(8): 579-82, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22672137

RESUMO

Domesticated adult dogs with antibody titer classified as below 'high' to one or more of canine distemper virus (CDV), canine parvovirus type-2 (CPV-2) and canine adenovirus type-1 (CAdV-1) were then given an additional inoculation, and the effectiveness of this booster evaluated 2 months later. Consequently, CDV and CAdV-1 antibody titer experienced a significant increase, but the same effect was not observed in the antibody titer of CPV-2. These findings suggest that with additional inoculation, a booster effect may be expected in increasing antibody titers for CDV and CAdV-1, but it is unlikely to give an increase in CPV-2 antibody titer.


Assuntos
Adenovirus Caninos/imunologia , Vírus da Cinomose Canina/imunologia , Imunização Secundária , Parvovirus Canino/imunologia , Vacinas Virais/imunologia , Animais , Anticorpos Antivirais/sangue , Cães , Vacinas Combinadas/administração & dosagem , Vacinas Combinadas/imunologia , Vacinas Virais/administração & dosagem
15.
Cancer Sci ; 103(8): 1546-52, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22587436

RESUMO

We previously reported that 3'-sulfoquinovosyl-1'-monoacylglycerol (SQMG) effectively suppresses the growth of solid tumors, likely via its anti-angiogenic activity. To investigate how SQMG affects angiogenesis, we performed DNA microarray analysis and quantitative real-time polymerase chain reaction. Consequently, upregulation of thrombospondin 1 (TSP-1) in SQMG-treated tumors in vitro and in vivo was confirmed. To address the mechanisms of TSP-1 upregulation by SQMG, we established stable TSP-1-knockdown transformants (TSP1-KT) by short hairpin RNA induction and performed reporter assay and in vivo assessment of anti-tumor assay. On the reporter assay, transcriptional upregulation of TSP-1 in TSP1-KT could not be induced by SQMG, thus suggesting that TSP-1 upregulation by SQMG occurred via TSP-1 molecule. In addition, growth of TSP1-KT xenografted tumors in vivo was not inhibited by SQMG, thus suggesting that anti-angiogenesis via TSP-1 upregulation induced by SQMG did not occur, as the SQMG target molecule TSP-1 was knocked down in TSP1-KT transformants. These data provide that SQMG is a promising candidate for the treatment of tumor-induced angiogenesis via TSP-1 upregulation.


Assuntos
Adenocarcinoma/metabolismo , Inibidores da Angiogênese/farmacologia , Neoplasias da Mama/metabolismo , Glicolipídeos/farmacologia , Monoglicerídeos/farmacologia , Trombospondina 1/metabolismo , Adenocarcinoma/genética , Inibidores da Angiogênese/genética , Animais , Neoplasias da Mama/genética , Linhagem Celular Tumoral , Perfilação da Expressão Gênica , Imuno-Histoquímica , Camundongos , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase em Tempo Real , Trombospondina 1/genética , Transcriptoma , Regulação para Cima/efeitos dos fármacos
16.
Can J Vet Res ; 76(4): 317-9, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23543958

RESUMO

The objective of this study was to determine whether post-vaccination antibody titers vary according to body weight in adult dogs. Antibody titers against canine parvovirus type 2 (CPV-2), canine distemper virus (CDV), and canine adenovirus type 1 (CAdV-1) were measured for 978 domestic adult dogs from 2 to 6 y of age. The dogs had been vaccinated approximately 12 mo earlier with a commercial combination vaccine. The dogs were divided into groups according to their weight. It was found that mean antibody titers in all weight groups were sufficient to prevent infection. Intergroup comparison, however, revealed that CPV-2 antibody titers were significantly higher in the Super Light (< 5 kg) group than in the Medium (10 to 19.9 kg) and Heavy (> 20 kg) groups and were also significantly higher in the Light (5 to 9.9 kg) group than in the Heavy group. Antibody titers against CDV were significantly higher in the Super Light, Light, and Medium groups than in the Heavy group. There were no significant differences among the groups for the CAdV-1 antibody titers.


Pour vérifier que les taux d'anticorps chez des chiens vaccinés changeaient en fonction de leur poids après la vaccination par un vaccin commercial combiné, on a mesuré les anticorps antivirus de la parvovirose canine (CPV-2), de la maladie de Carré (CDV) et de l'encéphalite de Rubarth ­ type-1 (CAdV-1) chez 978 chiens de compagnie agés de 2 à 6 ans, un an après leur vaccination. Par nos mesures, nous observons dans tous les groupes un taux satisfaisant d' immunisation moyen des animaux. Mais en comparant les groupes de poids, on s'aperçoit que pour la parvovirose canine CPV-2, le groupe des super-légers (< 5 kg) est significativement plus protégé en anticorps que les groupes de poids moyen (de 10 à 19,9 kg) et de poids le plus lourd (> 20 kg). De même les poids légers (de 5 à 9,9 kg) sont significativement mieux protégés que les poids lourds. Pour la maladie de Carré (CDV), les super-légers, les poids légers ou les groupes de poids moyen ont un taux d'anticorps significativement plus élevé que les plus lourds. Par contre pour l'Encéphalite de Rubarth (CAdV-1) aucune différence des taux d'anticorps dans les groupes de poids n'a été observée.(Traduit par les auteurs).


Assuntos
Adenovirus Caninos/imunologia , Anticorpos Antivirais/sangue , Vírus da Cinomose Canina/imunologia , Doenças do Cão/virologia , Parvovirus Canino/imunologia , Vacinas Virais/imunologia , Animais , Peso Corporal/imunologia , Doenças do Cão/imunologia , Doenças do Cão/prevenção & controle , Cães , Feminino , Masculino , Estatísticas não Paramétricas , Vacinação/veterinária , Vacinas Virais/administração & dosagem
17.
J Vet Diagn Invest ; 23(4): 832-5, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21908334

RESUMO

The current report describes a complex canine mammary adenoma with a rare histological feature characterized by sebaceous differentiation of tumor cells. A 13-year-old, mixed-breed, intact female dog had mammary tumors on the right mammary chain. Histologically, one of the masses was composed of bilayered ductal structures with luminal epithelial cells together with basaloid or myoepithelial cell components. Within the tumor, there were a number of lobules and nests of large foamy cells associated with basaloid reserve-like cells similar to sebaceous gland. Squamous metaplasia was also seen within the tumor. Immunohistochemical staining indicated that the tumor cells with sebaceous differentiation were positive for cytokeratin (CK)14 and that the associated basaloid reserve-like cells were positive for p63. In contrast, other luminal epithelial tumor cells were positive for CK18 and CK19, but not for CK14 and p63. The myoepithelial cells were positive for α-smooth muscle actin and p63. The expression of p63 in both sebaceous basaloid reserve-like cells and myoepithelial cells, and their structural continuity within the tumor tissue, suggested a common origin of these 2 components.


Assuntos
Adenoma/veterinária , Doenças do Cão/patologia , Neoplasias Mamárias Animais/patologia , Adenoma/diagnóstico , Adenoma/patologia , Adenoma/cirurgia , Animais , Doenças do Cão/diagnóstico , Doenças do Cão/cirurgia , Cães , Feminino , Neoplasias Mamárias Animais/diagnóstico , Neoplasias Mamárias Animais/cirurgia , Mastectomia/veterinária
18.
J Vet Diagn Invest ; 23(5): 1051-5, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21908374

RESUMO

A 9-year-old, female German Shepherd Dog presented with a firm bulging lesion at the tip of the tail. Histologically, the lesion was characterized by numerous poorly circumscribed clusters of vascular structures resembling capillaries that were separated by normal or mucinous connective tissues. The capillary-like vascular structures were composed of several layers: a single von Willebrand factor-positive endothelial cell layer with round, oval, or flattened hyperchromatic nuclei; and 1 or 2 surrounding spindle-shaped smooth muscle actin-positive pericyte layers. In the deep portion of the lesion, there were large vessels that showed morphological aberrations. These thin-to-thick-walled blood vessels corresponded to arteries and veins of varying diameters and were surrounded by fibrosis. The present case report describes a rare cutaneous vascular hamartoma, accompanied by aberrant arteriovenous structures.


Assuntos
Malformações Arteriovenosas/veterinária , Doenças do Cão/patologia , Hamartoma/veterinária , Cauda/patologia , Animais , Malformações Arteriovenosas/diagnóstico , Malformações Arteriovenosas/patologia , Doenças do Cão/diagnóstico , Cães , Feminino , Hamartoma/patologia
19.
PLoS One ; 6(4): e18285, 2011 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-21559518

RESUMO

BACKGROUND: Cyclosporin A (CsA) is well known as an immunosuppressive drug useful for allogeneic transplantation. It has been reported that CsA inhibits hepatitis C virus (HCV) genome replication, which indicates that cellular targets of CsA regulate the viral replication. However, the regulation mechanisms of HCV replication governed by CsA target proteins have not been fully understood. PRINCIPAL FINDINGS: Here we show a chemical biology approach that elucidates a novel mechanism of HCV replication. We developed a phage display screening to investigate compound-peptide interaction and identified a novel cellular target molecule of CsA. This protein, named CsA associated helicase-like protein (CAHL), possessed RNA-dependent ATPase activity that was negated by treatment with CsA. The downregulation of CAHL in the cells resulted in a decrease of HCV genome replication. CAHL formed a complex with HCV-derived RNA polymerase NS5B and host-derived cyclophilin B (CyPB), known as a cellular cofactor for HCV replication, to regulate NS5B-CyPB interaction. CONCLUSIONS: We found a cellular factor, CAHL, as CsA associated helicase-like protein, which would form trimer complex with CyPB and NS5B of HCV. The strategy using a chemical compound and identifying its target molecule by our phage display analysis is useful to reveal a novel mechanism underlying cellular and viral physiology.


Assuntos
Adenosina Trifosfatases/biossíntese , Ciclofilinas/metabolismo , Ciclosporina/farmacologia , RNA Polimerases Dirigidas por DNA/metabolismo , Hepacivirus/genética , Imunossupressores/farmacologia , RNA Viral/genética , Adenosina Trifosfatases/química , Adenosina Trifosfatases/fisiologia , Linhagem Celular Tumoral , DNA Complementar/metabolismo , Humanos , Biblioteca de Peptídeos , Plasmídeos/metabolismo , RNA Helicases , RNA Viral/metabolismo , Proteínas Recombinantes/química , Ressonância de Plasmônio de Superfície , Distribuição Tecidual
20.
Int Immunol ; 23(4): 223-37, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21421737

RESUMO

Extracellular heat shock protein can deliver associated antigens into the MHC class I presentation pathway of antigen-presenting cells, a process called cross-presentation, thus inducing antigen-specific CD8(+) T-cell responses; however, the precise mechanism for intracellular antigen translocation and the processing pathway has not been fully elucidated. Here we demonstrate that cross-presentation of extracellular Hsp90-ovalbumin (OVA) protein complexes to specific CD8(+) T cells involves both classical proteasome-transporter-associated antigen processing (TAP)-dependent and TAP-independent-endosomal pathways. Using confocal microscopy, we found that the internalized extracellular Hsp90 and OVA co-localized with cytosolic proteasomes. When anti-Hsp90 mAb was introduced to dendritic cells (DCs), we observed that the co-localization of internalized Hsp90-chaperoned OVA and proteasomes was abolished, resulting in the inhibition of TAP-dependent cross-presentation of OVA. Thus, extracellular Hsp90 may play a pivotal role for the translocation of chaperoned antigens for proteasomal degradation in the cytosol. In contrast, OVA chaperoned by Hsp90 was not presented by MHC class II molecules in vitro or in vivo, although the antigen was exogenously loaded onto DCs. Our data indicate that extracellular Hsp90 might be essential for the translocation of chaperoned antigens from the extracellular milieu into cytosol, resulting in proteasomal degradation for cross-presentation.


Assuntos
Antígenos/metabolismo , Células Dendríticas/metabolismo , Proteínas de Choque Térmico HSP90/metabolismo , Chaperonas Moleculares/metabolismo , Ovalbumina/metabolismo , Fragmentos de Peptídeos/metabolismo , Animais , Antígenos/imunologia , Linfócitos T CD8-Positivos/citologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Apresentação Cruzada , Citosol , Células Dendríticas/citologia , Células Dendríticas/imunologia , Endossomos/metabolismo , Espaço Extracelular , Proteínas de Choque Térmico HSP90/imunologia , Antígenos de Histocompatibilidade/metabolismo , Hibridomas , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Chaperonas Moleculares/imunologia , Ovalbumina/imunologia , Fragmentos de Peptídeos/imunologia , Complexo de Endopeptidases do Proteassoma/metabolismo , Ligação Proteica , Transporte Proteico
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