RESUMEN
MACC1 (metastasis associated in colon cancer 1) is a prognostic biomarker for tumor progression, metastasis and survival of a variety of solid cancers including colorectal cancer (CRC). Here we aimed to identify the MACC1-induced transcriptome and key players mediating the MACC1-induced effects in CRC. We performed microarray analyses using CRC cells ectopically overexpressing MACC1. We identified more than 1300 genes at least twofold differentially expressed, including the gene SPON2 (Spondin 2) as 90-fold upregulated transcriptional target of MACC1. MACC1-dependent SPON2 expression regulation was validated on mRNA and protein levels in MACC1 high (endogenously or ectopically) and low (endogenously or by knockdown) expressing cells. Chromatin immunoprecipitation analysis demonstrated the binding of MACC1 to the gene promoter of SPON2. In cell culture, ectopic SPON2 overexpression induced cell viability, migration, invasion and colony formation in endogenously MACC1 and SPON2 low expressing cells, whereas SPON2 knockdown reduced proliferative, migratory and invasive abilities in CRC cells with high endogenous MACC1 and SPON2 expression. In intrasplenically transplanted NOD/SCID mice, metastasis induction was analyzed with control or SPON2-overexpressing CRC cells. Tumors with SPON2 overexpression induced liver metastasis (vs control animals without any metastases, P=0.0036). In CRC patients, SPON2 expression was determined in primary tumors (stages I-III), and survival time was analyzed by Kaplan-Meier method. CRC patients with high SPON2 expressing primary tumors demonstrated 8 months shorter metastasis-free survival (MFS) compared with patients with low SPON2 levels (P=0.053). Combining high levels of SPON2 and MACC1 improved the identification of high-risk patients with a 20-month shorter MFS vs patients with low biomarker expression. In summary, SPON2 is a transcriptional target of the metastasis gene MACC1. SPON2 induces cell motility in vitro and CRC metastasis in mice. In patients, SPON2 serves as prognostic indicator for CRC metastasis and survival, and might represent a promising target for therapeutic approaches.
Asunto(s)
Biomarcadores de Tumor , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/metabolismo , Proteínas de la Matriz Extracelular/genética , Proteínas de Neoplasias/genética , Factores de Transcripción/metabolismo , Animales , Movimiento Celular/genética , Proliferación Celular , Supervivencia Celular/genética , Análisis por Conglomerados , Neoplasias Colorrectales/mortalidad , Neoplasias Colorrectales/patología , Biología Computacional/métodos , Modelos Animales de Enfermedad , Proteínas de la Matriz Extracelular/metabolismo , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Técnicas de Silenciamiento del Gen , Ontología de Genes , Xenoinjertos , Humanos , Ratones , Metástasis de la Neoplasia , Proteínas de Neoplasias/metabolismo , Estadificación de Neoplasias , Pronóstico , Unión Proteica , Curva ROC , TransactivadoresRESUMEN
Encephalitozoon intestinalis infection of sows is reported from a pig farm in Slovakia. Spores were detected by direct microscopic visualisation in the faeces of 25 out of 27 sows (92.6%). This finding was also supported serologically by the presence of specific anti-E. intestinalis antibodies and by a species-specific polymerase chain reaction (PCR). This is the first report on E. intestinalis infection of swine in Europe.
Asunto(s)
Encephalitozoon/aislamiento & purificación , Encefalitozoonosis/veterinaria , Enfermedades de los Porcinos/epidemiología , Animales , Anticuerpos Antifúngicos/sangre , Encephalitozoon/inmunología , Encefalitozoonosis/epidemiología , Ensayo de Inmunoadsorción Enzimática/métodos , Ensayo de Inmunoadsorción Enzimática/veterinaria , Europa (Continente)/epidemiología , Heces/microbiología , Femenino , Reacción en Cadena de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa/veterinaria , Eslovaquia/epidemiología , PorcinosRESUMEN
The effect of fungal and yeast glucan on different immune functions in mice was examined and compared. The simultaneous administration of glucan and a sensitizing dose of DNFB on the different sites significantly stimulated delayed-type hypersensitivity (DTH) response only when using fungal glucan. Both glucans tested, when administered before sensitization, significantly increased DTH response, but with a significantly higher level at the beginning of the investigation (on day 7) when using fungal glucan. The increase in phagocytic activity by the blood leucocytes started in the 1st week after fungal-glucan treatment, and in the 2nd week after yeast-glucan treatment, and took longer after administration of fungal glucan. The values of the phagocytic-activity index were significantly influenced only after fungal-glucan injection. The results of the study indicate that fungal glucan isolated from Pleurotus ostreatus could be a prospective immunomodulating substance.
Asunto(s)
Glucanos/farmacología , Hipersensibilidad Tardía/inmunología , Leucocitos/efectos de los fármacos , Fagocitosis/efectos de los fármacos , Polyporaceae/inmunología , Animales , Dinitrofluorobenceno/toxicidad , Hipersensibilidad a las Drogas/inmunología , Femenino , Glucanos/inmunología , Hipersensibilidad Tardía/inducido químicamente , Leucocitos/inmunología , Masculino , Ratones , Organismos Libres de Patógenos EspecíficosRESUMEN
The level of cell-mediated immune response in vivo was investigated using the test of delayed hypersensitive reaction (DHR) to DNFB, along with the phagocytic activity (PA) of blood leucocytes in mice after subcutaneous implantation of fungal and yeast glucan and levamisol in dependence on the dose and administration schedule. The soluble form of fungal glucan (Pleurotus ostreatus) potentiated the DHR significantly at a dose of 10 mg/kg (but not at a dose of 50 mg/kg) while it was administered during DNFB sensitization (P < 0.05)-Tab. I and when its pre-medication effect was investigated (days -7 and -14; P < 0.05) with regard to the time of sensitization (Tab. II). The identical dose of glucan also had a positive effect (P < 0.05 or 0.01) on the percentual proportion of phagocytic cells (PC) reaching the maximum in the 2nd and 3rd week of investigation, as well as on the phagocytic activity index (P < 0.05; 3rd week) and percentage of neutrophil granulocytes (P < 0.05; 2nd week)-Tab. III. Yeast glucan (Saccharomyces cerevisiae) showed a potentiating effect on the DHR to DNFB only in the case of its pre-medication use; its soluble form was effective at both doses (10 mg and 50 mg/kg) in days -7 and -14 (P < 0.05), and its corpuscular form at a dose of 50 mg/kg on days -7, -14 and -21 (P < 0.05 or 0.01)-Tab. II. PA parameters of blood leucocytes displayed a stimulative effect only on the PC percentage. The most significant effects in this case were observed in the soluble form (both doses) in the 2nd and 3rd week (P < 0.01 and 0.05, resp.) and in the insoluble form (both doses) in the 3rd and 4th week of observation (P < 0.05 and 0.01, resp.). An increase in the number of neutrophil granulocytes was significant in the 2nd (P < 0.05 or 0.01; corpuscular form) and 3rd week of the experiment (P < 0.01; soluble form)-Tab. III. Levamisol affected both investigated parameters (DHR and PA) only at a dose 20 mg/kg (10 mg/kg-no effect). Its potentiating effect on the DHR level was observed both for its administration at the time of sensitization (P < 0.05) and for its administration on days 7 (P < 0.05) and 14 (P < 0.01) before DNFB sensitization (Tabs. I and II). A statistically significant increase in PC was recorded in weeks 2, 3 and 4 (P < 0.05 or 0.01), a statistically significant increase in the number of neutrophil granulocytes in the 3rd week of investigation (P < 0.05). The phagocytic activity index was not affected.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Glucanos/farmacología , Hipersensibilidad Tardía , Leucocitos/inmunología , Levamisol/farmacología , Fagocitosis/efectos de los fármacos , Polyporaceae/metabolismo , Animales , Dinitrofluorobenceno , Inmunidad Celular/efectos de los fármacos , Ratones , Saccharomyces cerevisiae/metabolismoRESUMEN
Applicability of a skin test induced by dinitrofluorobenzene (DNFB) to quantification of the actual level of cellular immunity (CI) in vivo and its level after an experimental immunomodulation intervention were evaluated in two breeds (40 animals in each) of fattening bulls (10-11 months old). At the selected methodical procedure of intensity determination of the delayed type of hypersensitivity (DTH), its average value reached 4.5 +/- 1.5 mm in 80 animals, while in 77.5% of bulls its level ranged from 3.6 to 9.6 mm, in 18.7% from 2.0 to 3.5 mm and in 3.8% remained less than 2.0 mm. Evident expression of the reaction points to the possibility of application of the used methodical procedure of the skin test using DNFB to quantify the level of CI response in vivo in cattle. Percentual representation of animals according to the intensity of skin reaction (Tab. I) and concentration of total serum immunoglobulins (Cs-Ig) and serum IgG (Tab. II) indicates the different cellular and humoral state of animals in investigated breeds. This is also confirmed by the recorded average values of mentioned parameters which were significantly lower (P less than 0.01; or 0.05) in animals of the first breed (4.0 +/- 1.3 mm; 28.3 +/- 4.4 U ZST, 18.4 +/- 3.5 g.l-1) than in breed 2 (4.9 +/- 1.6 mm; 32.5 +/- 3.8 U ZST; 20.3 +/- 3.5 g.l-1). The animals of each breed were divided into four experimental groups with the approximately equal actual levels of DTH (Tab. III).(ABSTRACT TRUNCATED AT 250 WORDS)