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1.
Surg Today ; 43(2): 155-62, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22398718

RESUMO

PURPOSE: One of the typical modifications on the surface of cancer cells is sialylation of terminal carbohydrates. The expression of several types of sialylation of glycoconjugates was investigated in colorectal cancer. METHODS: The cancer tissue specimens obtained from 65 colorectal cancer patients were stained with sialic acid-binding lectins from Maackia amurensis (MAM), Sambucus sieboldiana (SSA), Maackia amurensis agglutinin (MAA) and monoclonal antibodies, and compared with their clinicopathological features. RESULTS: Cancer tissue specimens from 44.6% of patients had positive staining with MAM, which recognized α2,3sialylated type 2 chain (NeuAcα2,3Galß1,4GlcNAcßR) structures, but normal colorectal mucosa showed only weak staining with MAM was observed. More lymph node metastases and lymphatic invasion were seen in patients with positive staining with MAM (P < 0.01), while not with other lectins or antibodies that recognized sialylated glycoconjugates or sialyl Lewis-related antigens. The five-year survival rate of patients with MAM-positive staining was significantly lower than that with MAM-negative staining when including T0-1 cases, but there was no difference in cases with T2-4. There was no difference in the patients' survival rates when the tissues were stained with MAA, SSA or PNA lectins. CONCLUSION: α2,3Sialylated type 2 chain structures were predominantly expressed in colorectal tissues associated with the malignant transformation, in particular, with lymphatic spread of distal colorectal adenocarcinomas.


Assuntos
Adenocarcinoma/patologia , Biomarcadores Tumorais/metabolismo , Neoplasias Colorretais/patologia , Glicoconjugados/metabolismo , Ácidos Siálicos/metabolismo , Adenocarcinoma/metabolismo , Adenocarcinoma/mortalidade , Adenocarcinoma/cirurgia , Biomarcadores Tumorais/química , Transformação Celular Neoplásica/química , Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/patologia , Colectomia , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/mortalidade , Neoplasias Colorretais/cirurgia , Feminino , Glicoconjugados/química , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Metástase Linfática , Masculino , Lectinas de Plantas , Reto/cirurgia , Ácidos Siálicos/química , Taxa de Sobrevida , Resultado do Tratamento
2.
Clin Chim Acta ; 538: 181-188, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36403666

RESUMO

BACKGROUND: The antigenic determinant of CA19-9 is synthesized by the α1,3/4fucosyltransferase encoded by the Le gene in the Lewis blood group system. Accordingly, a diagnosis with CA19-9 is not appropriate forLe-negative patients who possess the Le gene-mutated le alleles homozygously. METHODS: A Le gene-specific PCR was undertaken to determine c59T>G by using a set of tag-sense and biotin-labeled anti-sense primers and a peptide nucleic acid-le-clamp which bound to G59 in the le alleles. Following mixing with streptavidin-coatedbluelatex beads, the PCR products were developed on a strip on which the complementary tag oligonucleotide to theLe gene-specific amplicon was immobilized. RESULTS: When the PCR products were developed on the strip, a clear line was rapidly observed in Le-positive but not in Le-negative individuals. In contrast, a significant number of cancer patients with Lewis-negative phenotype were found to possess CA19-9, while they were specifically genotyped asLe/-. No contradictory results were observed in cancer patients (n = 315) with respect to their Lewis genotypes and CA19-9 levels. CONCLUSIONS: c59T>G occurred commonly in the le alleles could be specifically and rapidly identified by the present method. This method appeared to be relevant forselecting cancer patientsto bediagnosed with CA19-9.


Assuntos
Antígeno CA-19-9 , Técnicas de Genotipagem , Neoplasias , Humanos , Antígeno CA-19-9/genética , Epitopos , Antígenos do Grupo Sanguíneo de Lewis/genética , Neoplasias/diagnóstico , Neoplasias/genética , Técnicas de Genotipagem/métodos
3.
Sci Rep ; 13(1): 4947, 2023 03 27.
Artigo em Inglês | MEDLINE | ID: mdl-36973299

RESUMO

A- and B-antigens are present on red blood cells (RBCs) as well as other cells and secretions in Hominoidea including humans and apes such as chimpanzees and gibbons, whereas expression of these antigens on RBCs is subtle in monkeys such as Japanese macaques. Previous studies have indicated that H-antigen expression has not completely developed on RBCs in monkeys. Such antigen expression requires the presence of H-antigen and A- or B-transferase expression in cells of erythroid lineage, although whether or not ABO gene regulation is associated with the difference of A- or B-antigen expression between Hominoidea and monkeys has not been examined. Since it has been suggested that ABO expression on human erythrocytes is dependent upon an erythroid cell-specific regulatory region or the + 5.8-kb site in intron 1, we compared the sequences of ABO intron 1 among non-human primates, and demonstrated the presence of sites orthologous to the + 5.8-kb site in chimpanzees and gibbons, and their absence in Japanese macaques. In addition, luciferase assays revealed that the former orthologues enhanced promoter activity, whereas the corresponding site in the latter did not. These results suggested that the A- or B-antigens on RBCs might be ascribed to emergence of the + 5.8-kb site or the corresponding regions in ABO through genetic evolution.


Assuntos
Hylobates , Pan troglodytes , Animais , Íntrons/genética , Pan troglodytes/genética , Hylobates/genética , Macaca fuscata , Sistema ABO de Grupos Sanguíneos/genética , Sistema ABO de Grupos Sanguíneos/metabolismo , Células Eritroides/metabolismo , Sequências Reguladoras de Ácido Nucleico , Eritrócitos/metabolismo , Primatas/genética , Antígenos/metabolismo
4.
J Hepatobiliary Pancreat Sci ; 30(4): 503-513, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35776060

RESUMO

BACKGROUND: Pancreatic intraductal papillary mucinous neoplasm (IPMN) involves multiple histopathological stages from benign to malignant lesions. Further, a biomarker to diagnose the malignant IPMN (IPMC) is clinically relevant. Recently, we found that serum fucosylated α1 -acid glycoprotein (fAGP) level markedly elevated along with disease progression in large cohorts of patients with various cancers. METHODS: The fAGP level was retrospectively analyzed in preoperative sera from 109 patients with IPMN, and the clinical relevance of fAGP was compared with currently available predictors as standard. RESULTS: The fAGP level in IPMC was found to be significantly higher than in benign IPMN (P = .0012). At a cutoff value of 27.04 U/µg, its sensitivity, specificity, and accuracy for IPMC were determined to be 83.61%, 65.96%, and 75.93%, respectively. Multivariate analyses revealed that the fAGP level was the only independent risk factor for predicting IPMC. Additionally, a combination of the fAGP level and 18 F-fluorodeoxyglucose uptake on the PET/CT imaging in the lesions seemed to offer the best diagnosis of IPMN. Accordingly, 27 of the 28 patients who were positive in both tests had IPMC, while patients who are negative had benign IPMN. CONCLUSIONS: The fAGP level appeared to be a relevant biomarker for malignant potential of IPMN.


Assuntos
Adenocarcinoma Mucinoso , Carcinoma Ductal Pancreático , Neoplasias Intraductais Pancreáticas , Neoplasias Pancreáticas , Humanos , Neoplasias Intraductais Pancreáticas/patologia , Carcinoma Ductal Pancreático/cirurgia , Orosomucoide , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Estudos Retrospectivos , Adenocarcinoma Mucinoso/patologia , Neoplasias Pancreáticas/cirurgia
5.
Electrophoresis ; 32(16): 2188-93, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21766474

RESUMO

α(1)-Acid glycoprotein (AGP) was previously shown to be a marker candidate of disease progression and prognosis of patients with malignancies by analysis of its glycoforms via lectins. Herein, affinity capillary electrophoresis of fluorescein-labeled AGP using lectins with the aid of laser-induced fluorescence detection was developed for quantitative evaluation of the fractional ratios of concanavalin A-reactive or Aleuria aurantia lectin-reactive AGP. Labeled AGP was applied at the anodic end of a fused-silica capillary (50 µm id, 360 µm od, 27 cm long) coated with linear polyacryloyl-ß-alanyl-ß-alanine, and electrophoresis was carried out for about 10 min in 60 mM 3-morpholinopropane-1-sulfonic acid-NaOH buffer (pH 7.35). Addition of the lectins to the anode buffer resulted in the separation of lectin-reactive glycoform peaks from lectin-non-reactive glycoform peaks. Quantification of the peak area of each group revealed that the percent of lectin-reactive AGP is independent of a labeling ratio ranging from 0.4 to 1.5 mol fluorescein/mol AGP, i.e. the standard deviation of 0.5% for an average of 59.9% (n=3). In combination with a facile procedure for micro-purification of AGP from serum, the present procedure, marking the reactivity of AGP with lectins, should be useful in determining the prognosis for a large number of patients with malignancies.


Assuntos
Concanavalina A/metabolismo , Eletroforese Capilar/métodos , Lectinas/metabolismo , Orosomucoide/análise , Biomarcadores/análise , Biomarcadores/química , Biomarcadores/metabolismo , Humanos , Orosomucoide/química , Orosomucoide/metabolismo
6.
Hepatogastroenterology ; 58(112): 1958-62, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22024067

RESUMO

BACKGROUND/AIMS: Tumor cells at high density are considered to be resistant to hyperthermia. Our objective in this study was to investigate hyperthermia sensitivity of clusters, cancer cell aggregation, compared with that of monolayer cells. METHODOLOGY: Colon carcinoma cells HT29 were cultured on poly 2-hydroxyethyl methacrylate-coated dishes for 7 days and the clusters were selected by a 40µm pore filter. To detect the cell reproductive potential, a colony formation assay was performed in HT29 cells from a monolayer and from clusters after exposure to cis-diamino-dichloroplatinum, fluorouracil and/or hyperthermia. Western blotting was used to analyze the induction of heat shock protein expression by hyperthermia. RESULTS: Histological findings of the clusters less than 400µm in diameter showed dividing cells and no secondary central necrosis. Cluster cells were more sensitive to hyperthermia than monolayer cells (p<0.0001). However, cluster formation induced cis-diamino-dichloroplatinum resistance (p<0.0001). The enhancement of hyperthermia sensitivity in clusters was not observed when the cells were heated after dispersion to single cells (p<0.0001). No difference of heat-induced HSP70/72 and HSP27 expression between cluster cells and monolayer cells was found. CONCLUSIONS: Cluster formation induced hyperthermia sensitivity, and cell-to-cell interaction in the clusters might enhance hyperthermia sensitivity.


Assuntos
Comunicação Celular , Neoplasias do Colo/terapia , Hipertermia Induzida , Agregação Celular , Neoplasias do Colo/patologia , Células HT29 , Humanos , Antígeno Ki-67/análise
7.
Glycobiology ; 20(10): 1251-8, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20542872

RESUMO

The phenomenon of accommodation in recipients of blood group ABO incompatible kidney transplantation (iKTx), in which grafts survive despite the presence of blood group A or B antigen in the graft and the presence of corresponding antibodies in the recipient's blood, is not uncommon. alpha1,3-N-Acetylgalactosaminyltransferase and alpha1,3galactosyltransferase associated with the synthesis of blood group A and B antigen (A and B enzymes), respectively, were measured by a highly specific enzyme-linked immunosorbent assay in the sera and transplanted tissues of patients who underwent an ABO iKTx. Allogeneic A and B enzymes were present in the sera and tissues as well as A and B antigens in the tissues for a long period, which hitherto have never been seen in recipients prior to an iKTx. However, activities of these enzymes in the sera after an iKTx decreased in patients who experienced a serious acute antibody-mediated rejection and disappeared in patients who had an unrepairable rejection, leading to graft loss without establishment of accommodation. Our observations on the presence of allogeneic A and B enzymes in the recipients' sera should have implications in decision making for a successful iKTx.


Assuntos
Sistema ABO de Grupos Sanguíneos/imunologia , Sistema ABO de Grupos Sanguíneos/metabolismo , Incompatibilidade de Grupos Sanguíneos/imunologia , Rejeição de Enxerto/imunologia , Sobrevivência de Enxerto/imunologia , Transplante de Rim/imunologia , Adulto , Idoso , Reações Antígeno-Anticorpo , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática , Galactosiltransferases/metabolismo , Humanos , Tolerância Imunológica , Técnicas Imunoenzimáticas , Falência Renal Crônica/terapia , Pessoa de Meia-Idade , N-Acetilgalactosaminiltransferases/metabolismo , Transplante Homólogo
8.
Sci Rep ; 9(1): 14503, 2019 10 10.
Artigo em Inglês | MEDLINE | ID: mdl-31601857

RESUMO

Immunotherapy targeting immune checkpoint molecules has provided remarkable clinical benefits in cancer patients but no clinically relevant biomarker for predicting treatment outcomes exists. Recently, we demonstrated that glycan structures of serum α1-acid glycoprotein (AGP) changed dramatically in cancer patients and that α1,3fucosylated AGP (fAGP) levels increased along with disease progression and decreased responding to chemotherapy treatments. Here, the fAGP was analyzed in sera prospectively obtained from 39 patients with advanced lung cancer who underwent immunotherapy with anti-PD-1 antibody, nivolumab. Twenty-three patients had significantly high fAGP levels above the cut-off value (H-fAGP) at one month after starting the treatment and 20 patients in this group, whose tumor sizes did not decrease, maintained high fAGP levels continuously and subsequently died. However, the other 16 patients, whose fAGP levels decreased or maintained below the cut-off value (L-fAGP), survived during a 2-year observation even though 5 patients in this group had no tumor shrinkage. Accordingly, the overall survival rate was found to significantly correlate with the fAGP level. Multivariate analyses revealed that the H-fAGP was an independent risk factor for cancer progression. Therefore, the fAGP level appeared to be a reliable biomarker for predicting clinical efficacy of immunotherapy with nivolumab.


Assuntos
Biomarcadores Tumorais/sangue , Neoplasias Pulmonares/terapia , Orosomucoide/genética , Prognóstico , Idoso , Biomarcadores Tumorais/genética , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glicosilação/efeitos dos fármacos , Humanos , Imunoterapia/métodos , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Nivolumabe/administração & dosagem
9.
Clin Chim Acta ; 478: 120-128, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29274330

RESUMO

BACKGROUND: Upon initiation and progression of cancer, α1-acid glycoprotein (AGP) possessing highly sialylated and fucosylated glycans appears in the serum, and recently has attracted a great deal of attention, as a potential biomarker of tumorigenesis in humans. METHODS: To establish a rapid and precise method for the quantitative assay of fucosylated AGP in serum samples, we developed an enzyme immunoassay (EIA) bearing an anti-AGP antibody and a fucose-binding lectin, Aleuria aurantia (AAL) with additional endeavor to improved sample handling, and antibody preparations. RESULTS: The amounts of fucosylated AGP could be determined by the present method with a good performance feature in all tested samples from both cancer patients and healthy controls. From cancer patients under chemotherapy we show that fucosylated AGP could be a clinically relevant biomarker for cancer progression or prognosis as well as for an early assessment of clinical response and treatment outcomes. Furthermore, in a different setting, fucosylated AGP also showed relevance in patients who received immunotherapy with an anti-programmed cell death-1 (PD-1) antibody. CONCLUSIONS: α1,3fucosylated AGP is a potential biomarker of cancer initiation, progression and response to treatment in cancer patients.


Assuntos
Biomarcadores Tumorais/sangue , Carcinogênese/química , Técnicas Imunoenzimáticas/métodos , Orosomucoide/metabolismo , Estudos de Casos e Controles , Progressão da Doença , Fucose/metabolismo , Glicosilação , Humanos , Ácido N-Acetilneuramínico/metabolismo , Neoplasias/diagnóstico , Neoplasias/patologia , Prognóstico
10.
PLoS One ; 11(6): e0156277, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27295180

RESUMO

One standard treatment option for advanced-stage cancer is surgical resection of malignant tumors following by adjuvant chemotherapy and chemoradiotherapy. Additionally, neoadjuvant chemotherapy may be applied if required. During the time course of treatments, patients are generally followed by computed tomography (CT) surveillance, and by tumor marker diagnosis. However, currently, early evidence of recurrence and/or metastasis of tumors with a clinically relevant biomarker remains a major therapeutic challenge. In particular, there has been no validated biomarker for predicting treatment outcomes in therapeutic settings. Recently, we have looked at glycoforms of serum α1-acid glycoprotein (AGP) by using a crossed affinoimmunoelectrophoresis with two lectins and an anti-AGP antibody. The primary glycan structures of AGP were also analyzed by a mass spectrometer and a novel software in a large number of patients with various cancers. Accordingly, the relative abundance of α1,3fucosylated glycans in AGP (FUCAGP) was found to be significantly high in cancer patients as compared with the healthy controls. Further, strikingly elevated levels of FUCAGP were found in patients with poor prognosis but not in patients with good prognosis. In the current study, levels of FUCAGP in serum samples from various cancer patients were analyzed and 17 patients including 13 who had undergone chemotherapy were followed for several years post operation. FUCAGP level determined diligently by using a mass spectrometer was found to change along with disease prognosis as well as with responses to treatments, in particular, to various chemotherapies. Therefore, FUCAGP levels measured during following-up of the patients after operation appeared to be clinically relevant biomarker of treatment intervention.


Assuntos
Biomarcadores Tumorais/sangue , Neoplasias/diagnóstico , Orosomucoide/análise , Polissacarídeos/sangue , Sequência de Carboidratos , Estudos de Casos e Controles , Fucose/metabolismo , Glicosilação , Humanos , Monitorização Fisiológica/métodos , Neoplasias/sangue , Neoplasias/cirurgia , Polissacarídeos/análise , Polissacarídeos/metabolismo , Período Pós-Operatório , Prognóstico , Análise de Sobrevida , Resultado do Tratamento
11.
Int J Oncol ; 47(6): 2255-63, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26498695

RESUMO

Curcuma zedoaria has been used as a traditional agent against malignant diseases. To elucidate detailed mechanisms producing such an activity, characterization and determination of molecular mechanisms of its antitumor effects was conducted. Inhibiting activities against cell proliferation, invasion and colony formation, and expression levels of corresponding molecules were investigated using human esophageal cancer TE-8 cells treated with the rhizome extract from C. zedoaria. Antitumor effect of the extract administered orally was also examined in tumor-bearing mice. The extract possessed strong anti-proliferation and invasion activities against TE-8 cells. Further, upregulated PTEN and downregulated phosphorylated Akt, mTOR and STAT3 expressions in the cells were induced shortly after treatment with the extract, followed by attenuation of FGFR1 and MMP-2, activation of caspase-9, caspase-3 and PARP, and suppression of Bcl-2 expressions, which led the cells to apoptotic cell death. Furthermore, tumor formation in mice was significantly suppressed through the oral administration of the extract. Taken together, these results suggest that the C. zedoaria extract could be a promising agent against esophageal cancer.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Carcinoma de Células Escamosas/patologia , Neoplasias Esofágicas/patologia , Extratos Vegetais/farmacologia , Animais , Apoptose/efeitos dos fármacos , Western Blotting , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Curcuma , Carcinoma de Células Escamosas do Esôfago , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Rizoma , Ensaios Antitumorais Modelo de Xenoenxerto
12.
Cancer Lett ; 180(2): 203-10, 2002 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-12175553

RESUMO

The thermal responses of mouse colorectal carcinoma cells were investigated in the wild type cells and the transfected cells with human FUT1 gene which encodes alpha 1,2fucosyltransferase. The heat sensitivity was observed to increase in the FUT1 gene transfected cells and the effect of hyperthermia at 44 degrees C on these cells was demonstrated to be significant (P<0.001) to the wild type cells even though no remarkable difference in the expression of the heat shock protein, Hsp70 was found in these cells. Thus the expression of alpha 1,2fucosylated antigens seemed to increase the heat sensitivity in mouse colorectal carcinoma cells.


Assuntos
Neoplasias Colorretais/terapia , Fucosiltransferases/fisiologia , Hipertermia Induzida , Animais , Neoplasias Colorretais/patologia , Citometria de Fluxo , Fucosiltransferases/genética , Proteínas de Choque Térmico HSP70/biossíntese , Humanos , Camundongos , Transfecção , Galactosídeo 2-alfa-L-Fucosiltransferase
13.
J Neurosurg ; 120(6): 1298-308, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24678780

RESUMO

OBJECT: Glioblastoma is the most aggressive malignant brain tumor, and overall patient survival has not been prolonged even by conventional therapies. Previously, the authors found that chemically synthesized glycans could be anticancer agents against growth of a series of cancer cells. In this study, the authors examined the effects of glycans on the growth of glioblastoma cells both in vitro and in vivo. METHODS: The authors investigated not only the occurrence of changes in the cell signaling molecules and expression levels of various proteins related to cell death, but also a mouse model involving the injection of glioblastoma cells following the administration of synthetic glycans. RESULTS: Synthetic glycans inhibited the growth of glioblastoma cells, induced the apoptosis of the cells with cleaved poly (adenosine diphosphate-ribose) polymerase (PARP) expression and DNA fragmentation, and also caused autophagy, as shown by the detection of autophagosome proteins and monodansylcadaverine staining. Furthermore, tumor growth in the in vivo mouse model was significantly inhibited. A dramatic induction of programmed cell death was found in glioblastoma cells after treatment with synthetic glycans. CONCLUSIONS: These results suggest that synthetic glycans could be a promising novel anticancer agent for performing chemotherapy against glioblastoma.


Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Glioblastoma/metabolismo , Glioblastoma/patologia , Polissacarídeos/farmacologia , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Proteínas Reguladoras de Apoptose/genética , Neoplasias Encefálicas/tratamento farmacológico , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Colestanóis/farmacologia , Colestanóis/uso terapêutico , Fragmentação do DNA/efeitos dos fármacos , Modelos Animais de Doenças , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glioblastoma/tratamento farmacológico , Humanos , Técnicas In Vitro , Camundongos , Camundongos Nus , Poli(ADP-Ribose) Polimerases/metabolismo , Polissacarídeos/uso terapêutico , Resultado do Tratamento , Ensaios Antitumorais Modelo de Xenoenxerto
14.
PLoS One ; 9(2): e89071, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24558470

RESUMO

Blood group-related glycans determining ABO and Lewis blood groups are known to function as attachment factors for most of the norovirus (NoV) strains. To identify binding specificity of each NoV, recombinant norovirus-like particles (VLPs) and human saliva samples with different ABO, Lewis phenotypes and secretor status have been commonly applied. When binding specificities of VLPs prepared from 16 different genotypes of NoVs in GI and GII genogroups were characterized in samples of human gastric mucosa compared to human saliva based on blood group phenotypes, considerable differences were observed for several strains. Novel binding specificities determined by an ELISA using preparations from human gastric mucosa were also ascertained by immunohistochemical analyses using human jejunal mucosa, widely believed to be susceptible to NoV infection. Further, A, B and O(H) blood group substances prepared from porcine and squid tissues were found to be effective for preventing ABO blood group-specific binding of VLPs to both saliva and mucosa samples. Therefore, these blood group substances might have potential for the prevention and treatment of NoV infection.


Assuntos
Antígenos de Grupos Sanguíneos/metabolismo , Infecções por Caliciviridae/sangue , Infecções por Caliciviridae/prevenção & controle , Norovirus/metabolismo , Polissacarídeos/metabolismo , Animais , Antígenos de Grupos Sanguíneos/uso terapêutico , Decapodiformes , Ensaio de Imunoadsorção Enzimática , Mucosa Gástrica/metabolismo , Humanos , Imuno-Histoquímica , Polissacarídeos/sangue , Ligação Proteica/fisiologia , Saliva/metabolismo , Suínos , Vírion/metabolismo
15.
Biomed Res Int ; 2013: 834790, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23509786

RESUMO

Human plasma α1-acid glycoprotein (AGP) from cancer patients and healthy volunteers was purified by sequential application of ion-exchange columns, and N-linked glycans enzymatically released from AGP were labeled and applied to a mass spectrometer. Additionally, a novel software system for use in combination with a mass spectrometer to determine N-linked glycans in AGP was developed. A database with 607 glycans including 453 different glycan structures that were theoretically predicted to be present in AGP was prepared for designing the software called AGPAS. This AGPAS was applied to determine relative abundance of each glycan in the AGP molecules based on mass spectra. It was found that the relative abundance of fucosylated glycans in tri- and tetra-antennary structures (FUCAGP) was significantly higher in cancer patients as compared with the healthy group (P < 0.001). Furthermore, extremely elevated levels of FUCAGP were found specifically in patients with a poor prognosis but not in patients with a good prognosis. In conclusion, the present software system allowed rapid determination of the primary structures of AGP glycans. The fucosylated glycans as novel tumor markers have clinical relevance in the diagnosis and assessment of cancer progression as well as patient prognosis.


Assuntos
Fucose/química , Espectrometria de Massas/métodos , Neoplasias/sangue , Neoplasias/diagnóstico , Orosomucoide/análise , Biomarcadores Tumorais/sangue , Estudos de Casos e Controles , Cromatografia por Troca Iônica , Progressão da Doença , Ensaio de Imunoadsorção Enzimática , Glicosilação , Humanos , Plasma/metabolismo , Polissacarídeos/química , Prognóstico , Software
16.
Leg Med (Tokyo) ; 13(1): 22-9, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21144789

RESUMO

Gene expression is driven by promoters, enhancers, silencers, and other cis-regulatory elements upstream and downstream of the gene. Previous studies of the regulation of human ABO gene transcription have focused mainly on the 5' region, including the core promoter and the region proximal to it. However, as the involvement of the 3' flanking region in transcriptional regulation has not yet been examined, we focused on this issue. The 3' region approximately 2.2kb downstream of the ABO gene was PCR-amplified and inserted into a cloning vector, followed by sequence determination and preparation of luciferase reporter vectors. Transient transfections into KATOIII and K562 cells were performed using various reporter plasmids containing the 3' region. The 3' region of the ABO gene, which was characterized by a high degree of sequence repetition, was effectively cloned by a single-copy cloning method. Transfections in KATOIII and K562 cells showed that negative elements were demonstrable within the 3' region. These observations suggest that negative regulatory elements seem to be present in the 3' region of ABO in both epithelial and erythroid lineages. As we had observed a negative region just upstream of the ABO promoter, transcription from ABO could be negatively regulated by repressive regions just upstream of the promoter and downstream of the gene. Further studies of the enhancer will be required for elucidating the molecular basis of ABO gene expression.


Assuntos
Sistema ABO de Grupos Sanguíneos/genética , Regulação da Expressão Gênica/genética , Expressão Gênica/genética , Leucemia Mieloide/sangue , Regiões Promotoras Genéticas/genética , Neoplasias Gástricas/sangue , Sequência de Bases , Humanos , Luciferases/genética , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Fatores de Transcrição/genética
17.
Glycoconj J ; 25(6): 531-44, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18327639

RESUMO

Chemically synthesized sugar-cholestanols with mono-, di-, and tri-saccharides attached to cholestanol showed strong inhibiting activity against the proliferation of colorectal and gastric cancer cells. In contrast, cholestanol without sugar moieties was totally ineffective. Furthermore, when cancer cells were exposed to GlcNAcRbetacholestanol (R=(-) or beta1-3Gal), the compound was rapidly taken up via the lipid rafts/microdomains on the cell surface. The uptake of sugar-cholestanol in mitochondria increased gradually and was followed by the release of cytochrome c from mitochondria and the activation of apoptotic signals through the mitochondrial pathway and the caspase cascade, leading to apoptotic cell death, characterized by DNA ladder formation and nuclear fragmentation. Additionally, the examination of GlcNAcRbetacholestanol in a mouse model of peritoneal dissemination showed a dramatic reduction of tumor growth (P < 0.003) and prolonged mouse survival time (P<0.0001). Based on these observations, we believe that the sugar-cholestanols described here have clinical potential as novel anticancer agents.


Assuntos
Antineoplásicos/farmacologia , Colestanóis/farmacologia , Neoplasias Experimentais/tratamento farmacológico , Oligossacarídeos/farmacologia , Animais , Antineoplásicos/síntese química , Western Blotting , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Colestanóis/síntese química , Cromatografia Líquida , Neoplasias Colorretais/tratamento farmacológico , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Citocromos c/metabolismo , Fragmentação do DNA/efeitos dos fármacos , Feminino , Células HT29 , Humanos , Injeções Intraperitoneais , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias/métodos , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia , Oligossacarídeos/síntese química , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Espectrometria de Massas em Tandem
18.
Biol Pharm Bull ; 30(3): 588-90, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17329863

RESUMO

The adherence to HeLa cells by the yeast-type cells of the Candida albicans NIH A-207 strain cultivated for 2 d at 27 degrees C in the yeast extract-added Sabouraud liquid medium (YSLM) and the 500 mM galactose-added yeast nitrogen base medium (YNB-Gal) was compared. The yeast cells cultured in the YNB-Gal clearly increased the adherence to the HeLa cells compared to the YSLM. The adherence drastically decreased by pronase treatment of the yeast cells. Next, to define the sugar receptors of the yeast cells, lactosylceramide (LC)-, the H type 1 antigen (HA)-, Lewis(a) antigen (Le(a))-, mannan- and BSA-coated polystyrene beads were used for the adherence to the yeast cells. Only the LC- and HA-beads were bound to the yeast cells. The adherence of the two beads to the yeast cells cultured in the YNB-Gal was higher than that to the yeast cells cultured in the YSLM. The yeast cell wall mannan-coated polystyrene beads did not adhere at all to the Hela cells. Based on these results, it is evident that the protein parts involving the LC and HA receptors on the surface of the yeast cells correlate with the adherence to the HeLa cells.


Assuntos
Candida albicans/metabolismo , Microesferas , Poliestirenos/química , Antígenos CD/química , Antígenos CD/metabolismo , Antígenos de Diferenciação/química , Antígenos de Diferenciação/metabolismo , Candida albicans/citologia , Adesão Celular/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Galactose/química , Galactose/metabolismo , Células HeLa , Humanos , Lactosilceramidas/química , Lactosilceramidas/metabolismo , Antígenos do Grupo Sanguíneo de Lewis/química , Antígenos do Grupo Sanguíneo de Lewis/metabolismo , Glicoproteínas de Membrana/química , Poliestirenos/metabolismo , Pronase/química , Pronase/metabolismo , Albumina Sérica/química , Albumina Sérica/metabolismo
19.
Ann Surg Oncol ; 14(2): 960-7, 2007 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17094023

RESUMO

BACKGROUND: Tumor markers whose antigenic determinants have been demonstrated to consist of carbohydrates are probably one of the most extensive tools that have been used in routine cancer diagnosis. In this study, the relevance of carbohydrate antigen expression profile was examined in esophageal squamous cell carcinoma together with prognosis in 130 patients. METHODS: The expression of carbohydrate antigens was estimated immunohistochemically by anti-sialyl Lewis a (sialyl Le(a)) and anti-sialyl Lewis x (sialyl Le(x)) monoclonal antibodies, and correlation between their staining and clinicopathological status was examined. In addition, the correlation of both carbohydrate antigens expression was evaluated with microvessel density (MVD). RESULTS: Expressions of sialyl Lewis antigens and MVD were associated with several clinicopathological features that reflect the tumor aggressiveness in esophageal cancer. The 5-year survival rate of patients was found to be associated with expression of sialyl Le(a) and sialyl Le(x) antigens and with MVD; thus, all of them were revealed to be independent prognostic factors. CONCLUSIONS: Combination of these factors offered a better prediction of prognosis of esophageal squamous cell carcinoma. Further, carbohydrate antigens represent a promising target for therapeutic approaches against the disease.


Assuntos
Carcinoma de Células Escamosas/metabolismo , Neoplasias Esofágicas/metabolismo , Gangliosídeos/biossíntese , Neovascularização Patológica/patologia , Oligossacarídeos/biossíntese , Idoso , Biomarcadores Tumorais , Antígeno CA-19-9 , Carcinoma de Células Escamosas/patologia , Neoplasias Esofágicas/patologia , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Prognóstico , Antígeno Sialil Lewis X , Análise de Sobrevida
20.
Ann Surg Oncol ; 14(6): 1880-9, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17375356

RESUMO

BACKGROUND: We have previously demonstrated tumor-specific alpha1,2fucosylation, which is associated with resistance of tumor cells to anticancer treatment in human colorectal tumor tissues. By using the YB-2 monoclonal antibody, the resulting products have been identified as Y, Le(b), and H type 2 antigens in colorectal tumor tissues. METHODS: Immunohistochemical analyses of colorectal cancer tissues (74 specimens) were performed with a newly established mouse monoclonal antibody, YB-3 specifically recognizing H disaccharide (Fucalpha1,2Galbeta) structures, and anti-A, anti-B, YB-2, and anti-sialyl Lewis X (SLX) antibodies, together with the analyses of glycosyltransferases involved in the synthesis of ABH antigens in the same tissues. RESULTS: The YB-3 antibody enabled us to detect colorectal tumors, particularly tumors in the distal large intestine and the rectum, with high sensitivity (74.3%) and specificity (100%). From immunohistochemical and enzymatic analyses of colorectal tissues, we found that once alpha1,2fucosylation had proceeded in tumor tissues, blood group A or B antigen was also synthesized in approximately half of the tissues of A or B blood type, but not in their normal tissues. A correlation of survival rate with immunostaining of tissues was found only by YB-3 antibody and not by anti-A, anti-B, or anti-SLX antibody. CONCLUSIONS: As a predictor of postoperative prognosis of patients with colorectal cancer, immunodetection of alpha1,2fucosylated antigens with the YB-3 antibody seemed to be superior to blood groups A, B, or SLX antigen in colorectal tumor tissues.


Assuntos
Sistema ABO de Grupos Sanguíneos/análise , Antígenos de Neoplasias/análise , Biomarcadores Tumorais/análise , Neoplasias do Colo/patologia , Antígenos do Grupo Sanguíneo de Lewis/análise , Antígenos CD15/análise , Oligossacarídeos/análise , Neoplasias Retais/patologia , Anticorpos Monoclonais , Neoplasias do Colo/cirurgia , Dissacarídeos/análise , Feminino , Previsões , Fucosil Galactose alfa-N-Acetilgalactosaminiltransferase/análise , Glicosiltransferases/análise , Antígenos H-2 , Humanos , Masculino , Prognóstico , Neoplasias Retais/cirurgia , Antígeno Sialil Lewis X
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