RESUMEN
BACKGROUND: Apiole was isolated from the leaves of various plants and vegetables and has been demonstrated to inhibit human colon cancer cell (COLO 205 cells) growth through induction of G0/G1 cell cycle arrest and apoptotic cell death. This study further explored the antitumor effects of apiole derivatives AP-02, 04, and 05 in COLO 205 cancer cells. METHODS: Human breast (MDA-MB-231, ZR75), lung (A549, PE089), colon (COLO 205, HT 29), and hepatocellular (Hep G2, Hep 3B) cancer cells were treated with apiole and its derivatives in a dose-dependent manner. Flow cytometry analysis was subsequently performed to determine the mechanism of AP-02-induced G0/G1 cell cycle arrest. The in vivo antitumor effect of AP-02 (1 and 5 mg/kg, administered twice per week) was examined by treating athymic nude mice bearing COLO 205 tumor xenografts. The molecular mechanisms of AP-02-induced antitumor effects were determined using western blot analysis. RESULTS: AP-02 was the most effective compound, especially for inhibition of COLO 205 colon cancer cell growth. The cytotoxicity of AP-02 in normal colon epithelial (FHC) cells was significantly lower than that in other normal cells derived from the breast, lung or liver. Flow cytometry analysis indicated that AP-02-induced G0/G1 cell cycle arrest in COLO 205 cells but not in HT 29 cells (< 5 µM for 24 h, **p < 0.01). Tumor growth volume was also significantly inhibited in AP-02 (> 1 mg/kg)-treated athymic nude mice bearing COLO 205 tumor xenografts compared to control mice (*p < 0.05). Furthermore, G0/G1 phase regulatory proteins (p53 and p21/Cip1) and an invasion suppressor protein (E-cadherin) were significantly upregulated, while cyclin D1 was significantly downregulated, in AP-02-treated tumor tissues compared to the control group (> 1 mg/kg, *p < 0.05). CONCLUSIONS: Our results provide in vitro and in vivo molecular evidence of AP-02-induced anti-proliferative effects on colon cancer, indicating that this compound might have potential clinical applications.
Asunto(s)
Antineoplásicos/administración & dosificación , Neoplasias del Colon/tratamiento farmacológico , Dioxoles/administración & dosificación , Puntos de Control de la Fase G1 del Ciclo Celular/efectos de los fármacos , Petroselinum/química , Animales , Antineoplásicos/efectos adversos , Antineoplásicos/química , Apoptosis/efectos de los fármacos , Neoplasias del Colon/fisiopatología , Ciclina D1/genética , Ciclina D1/metabolismo , Dioxoles/efectos adversos , Dioxoles/química , Femenino , Humanos , Ratones , Ratones Desnudos , Fase de Descanso del Ciclo Celular/efectos de los fármacos , Proteína p53 Supresora de Tumor/genética , Proteína p53 Supresora de Tumor/metabolismo , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Human triple-negative breast cancer (TNBC) is the most aggressive and poorly understood subclass of breast cancer. Glucose transporters (GLUTs) are required for glucose uptake in malignant cancer cells and are ideal targets for cancer therapy. To determine whether the inhibition of GLUTs could be used in TNBC cell therapy, the apple polyphenol phloretin (Ph) was used as a specific antagonist of GLUT2 protein function in human TNBC cells. Interestingly, we found that Ph (10-150 µM, for 24 h) inhibited cell growth and arrested the cell cycle in MDA-MB-231 cells in a p53 mutant-dependent manner, which was confirmed by pre-treatment of the cells with a p53-specific dominant-negative expression vector. We also found that Ph treatment (10-150 µM, for 24 h) significantly decreased the migratory activity of the MDA-MB-231 cells through the inhibition of paxillin/FAK, Src, and alpha smooth muscle actin (α-sMA) and through the activation of E-cadherin. Furthermore, the anti-tumorigenic effect of Ph (10, 50 mg/kg or DMSO twice a week for six weeks) was demonstrated in vivo using BALB/c nude mice bearing MDA-MB-231 tumor xenografts. A decrease in N-cadherin, vimentin and an increase in p53, p21 and E-cadherin were detected in the tumor tissues. In conclusion, inhibition of GLUT2 by the apple polyphenol Ph could potentially suppress TNBC tumor cell growth and metastasis.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Movimiento Celular/efectos de los fármacos , Transportador de Glucosa de Tipo 2/metabolismo , Malus/química , Floretina/farmacología , Extractos Vegetales/farmacología , Transducción de Señal/efectos de los fármacos , Animales , Antineoplásicos Fitogénicos/química , Neoplasias de la Mama/metabolismo , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Femenino , Humanos , Ratones , Floretina/química , Extractos Vegetales/química , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Static broth culture favors Salmonella enterica subsp. enterica serovar Typhimurium to produce type 1 fimbriae, while solid agar inhibits its expression. A transposon inserted in stbC, which would encode an usher for Stb fimbriae of a non-flagellar Salmonella enterica subsp. enterica serovar Typhimurium LB5010 strain, conferred it to agglutinate yeast cells on both cultures. RT-PCR revealed that the expression of the fimbrial subunit gene fimA, and fimZ, a regulatory gene of fimA, were both increased in the stbC mutant when grown on LB agar; fimW, a repressor gene of fimA, exhibited lower expression. Flagella were observed in the stbC mutant and this phenotype was correlated with the motile phenotype. Microarray data and RT-PCR indicated that the expression of three genes, motA, motB, and cheM, was enhanced in the stbC mutant. The stbC mutant was resistant to several antibiotics, consistent with the finding that expression of yhcQ and ramA was enhanced. A complementation test revealed that transforming a recombinant plasmid possessing the stbC restored the mannose-sensitive agglutination phenotype to the stbC mutant much as that in the parental Salmonella enterica subsp. enterica serovar Typhimurium LB5010 strain, indicating the possibility of an interplay of different fimbrial systems in coordinating their expression.
Asunto(s)
Proteínas Bacterianas/metabolismo , Elementos Transponibles de ADN/genética , Fimbrias Bacterianas/genética , Fimbrias Bacterianas/metabolismo , Manosa/farmacología , Salmonella typhimurium/genética , Salmonella typhimurium/metabolismo , Antígenos Bacterianos/genética , Antígenos Bacterianos/metabolismo , Proteínas Bacterianas/genética , Elementos Transponibles de ADN/fisiología , Proteínas Fimbrias/genética , Proteínas Fimbrias/metabolismo , Fimbrias Bacterianas/efectos de los fármacos , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Regulación Bacteriana de la Expresión Génica/genética , Fenotipo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Salmonella typhimurium/efectos de los fármacosRESUMEN
BACKGROUND: Reactive thrombocytosis (RT) in pediatric patients is common, but usually without symptoms. The incidence of RT is different depending on age. Mostly, we reason that RT is a phenomenon, nevertheless the diagnostic value of RT is little known. Therefore, the aim of this study was to determine the association of RT and clinical or laboratory characteristics in pediatric diseases. METHODS: We retrospectively analyzed the medical records of pediatric patients hospitalized at Wan Fang hospital from January 2002 to July 2009. Thrombocytosis was defined as a platelet count more than 500 × 10(9)/L. There were 822 patients enrolled to this study. The clinical parameters, including age, gender, disease type, and hospitalization days, were investigated. The association between RT and clinical manifestations and the relationship of leukocytes, hemoglobin, C-reactive protein, and platelet counts were analyzed. RESULTS: The overall incidence of RT in hospitalized pediatric patients was 6.3%. Infants had a significantly higher incidence (11.3%, p<0.001). Mild RT was found in most patients (83.6%). Infections (75.4%) were the most common cause, followed by perinatal diseases (11.1%). The relationship of RT and age revealed a positive correlation (p=0.045, r=0.70 after adjustment). The degree of RT was an independent factor for hospitalization days (p<0.001, r=0.126 after adjustment). There was a positive correlation between white blood cell count and platelets (p=0.002, r=0.017); on the contrary, the relationship between hemoglobin level and platelets was an inverse correlation (p<0.001, r=-0.193). CONCLUSIONS: In children, the degree of RT was associated with age, and patients had significantly longer hospitalization days in proportion to the increase in platelet count. Laboratory association revealed that the degree of RT was positively correlated to white cell count and negatively correlated to hemoglobin level. Therefore, the degree of RT might be a predictive factor with regard to hospitalization days in pediatric diseases.
Asunto(s)
Trombocitosis/fisiopatología , Adolescente , Factores de Edad , Niño , Preescolar , Femenino , Hemoglobinas/análisis , Humanos , Lactante , Recién Nacido , Infecciones/complicaciones , Tiempo de Internación , Recuento de Leucocitos , Masculino , Recuento de Plaquetas , Estudios RetrospectivosRESUMEN
Lead (Pb) may alter T-lymphocyte reactivity in situ by preferentially enhancing the development of T-helper 2 (T(H)2)- and inhibiting T(H)1-lymphocyte development. These effects could result in dysregulation of the presence/availability of T(H)1- and T(H)2-associated cytokines. The aim of this study was two-fold, that is, to assess whole blood Pb levels in schoolchildren from Taiwanese communities that varied in degree of potential for Pb exposure and then ascertain if there were relationships between Pb exposure and changes in levels of key T(H)1 and T(H)2 cytokines. Grades 5 and 6 students were selected from four different community schools, i.e., one from: urban area with new homes; urban area with old homes; rural site with old homes; and area located near an oil refinery. Students at each site were further divided into healthy and respiratory allergy subgroups. Blood was collected and whole blood Pb levels and serum interferon (IFN)-γ, interleukin (IL)-12, -4, and -5 levels were determined. The results indicate no differences in whole blood Pb levels (<4 µg/dl) among students from urban and rural sites; these values were similar in the healthy and allergic subjects. Serum T(H)1 and T(H)2 cytokine levels also did not differ among/within the groups. In contrast, refinery children had significantly increased Pb levels (5.2-8.8 µg/dl) relative to any of the other sets' levels. Of these, children with allergies had serum T(H)2 cytokine levels significantly higher and T(H)1 cytokine levels significantly lower than their healthy counterparts. Oddly, though having elevated Pb levels, healthy refinery students did not display altered T(H)1 or T(H)2 cytokine levels relative to control student values. From this, we conclude that substantively increased whole blood Pb levels may promote T(H) cell dysregulation and alter the availability of key T(H)1 and T(H)2 cytokines, effects that could ultimately contribute to development of pulmonary allergic diseases.
Asunto(s)
Exposición a Riesgos Ambientales/efectos adversos , Interleucinas/sangre , Intoxicación por Plomo/sangre , Plomo/efectos adversos , Células TH1/efectos de los fármacos , Células Th2/efectos de los fármacos , Asma/sangre , Industria Química , Niño , Industria Procesadora y de Extracción , Humanos , Interferón gamma/sangre , Plomo/sangre , Intoxicación por Plomo/epidemiología , Petróleo , Células TH1/metabolismo , Células Th2/inmunologíaRESUMEN
Cerebellar disorder was frequently reported to have relation with structural brain volume alteration and/or morphology change. In dealing with such clinical situations, we need a convenient and noninvasive imaging tool to provide clinicians with a means of tracing developmental changes in the cerebellum. Herein, we present a new daily practice method for cerebellum imaging that uses a work station and a software program to process reconstructed 3D neuroimages after MRI scanning. In a 3-y period, 3D neuroimages reconstructed from MRI scans of 50 children aged 0.2-12.7 y were taken. The resulting images were then statistically analyzed against a growth curve. We observed a remarkable increase in the size of the cerebellum in the first 2 y of life. Furthermore, the unmyelinated cerebellum grew mainly between birth and 2 y of age in the postnatal stage. In contrast, the postnatal development of the brain mainly depended on the growth of myelinated cerebellum from birth through adolescence. This study presents basic data from a study of ethnic Chinese children's cerebellums using reconstructed 3D brain images. Based on the technique we introduce here, clinicians can evaluate the growth of the brain.
Asunto(s)
Pueblo Asiatico , Cerebelo/anatomía & histología , Cerebelo/crecimiento & desarrollo , Imagenología Tridimensional/métodos , Imagen por Resonancia Magnética/métodos , Factores de Edad , Niño , Preescolar , Femenino , Humanos , Lactante , Estudios Longitudinales , Masculino , Tamaño de los Órganos/fisiología , TaiwánRESUMEN
OBJECTIVE: The three-dimensional (3D) reconstructed neuroimages are currently available to analyze brain structure. It provides a new tool for clinical evaluation and academic research on brain. However, there are several methods for processing 3D images. In this article, we present a technique that utilizes a work station and a software program to process reconstructed 3D neuroimages after magnetic resonance imaging (MRI) scanning. METHODS: The brain volumes of 50 normal children aged between 3 months and 12 years and 11 months were measured by 3D neuroimages reconstructed from regular MRI scans. These results were then analyzed statistically against the growth curve. RESULTS: The regression curve of cortical growth was y = 39.317Ln(x) + 631.31, R (2) = 0.1318. The regression curve of white matter growth was y = 81.754Ln(x) + 186.07, R(2) = 0.5675. The regression curve of whole brain growth was y = 121.07Ln(x) + 817.738, R (2) = 0.4077. Current studies show that at the postnatal stage, the cortex grows mainly between birth and 4 years of age. At the same time, the postnatal development of the brain depends mainly on the growth of white matter from birth through adolescence. CONCLUSIONS: This study presents the basic data from a study of children's brains using reconstructed 3D brain images. A 3D reconstructed neuroimage provides a new tool for neurological and psychological in vivo research of the brain. Based on the techniques we introduce here, the clinician may evaluate the growth of the brain in a more efficient and precise manner.
Asunto(s)
Encéfalo/crecimiento & desarrollo , Procesamiento de Imagen Asistido por Computador , Imagenología Tridimensional , Imagen por Resonancia Magnética , Corteza Cerebral/crecimiento & desarrollo , Niño , Preescolar , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Tamaño de los Órganos/fisiología , Valores de Referencia , Programas InformáticosRESUMEN
Cd is an industrial and environmental pollutant that affects many organs in humans and other mammals. However, the molecular mechanisms of Cd-induced nephrotoxicity are unclear. In this study, we show that endoplasmic reticula (ER) played a pivotal role in Cd-induced apoptosis in mesangial cells. Using Fluo-3 AM, the intracellular concentration of calcium ([Ca(2+)](i)) was detected as being elevated as time elapsed after Cd treatment. Co-treatment with BAPTA-AM, a calcium chelator, was able to significantly suppress Cd-induced apoptosis. Calcineurin is a cytosolic phosphatase, which was able to dephosphorylate the inositol-1,4,5-triphosphate receptor (IP(3)R) calcium channel to prevent the release of calcium from ER. Cyclosporine A, a calcineurin inhibitor, increased both [Ca(2+)](i) and the percentage of Cd-induced apoptosis. However, EGTA and the IP(3)R inhibitor, 2-APB, were able to partially modulate Cd cytotoxicity. These results led us to suggest that the extracellular and ER-released calcium plays a crucial role in Cd-induced apoptosis in mesangial cells. Following this line, we further detected the ER stress after Cd treatment since ER is one of the major calcium storage organelles. After Cd exposure, GADD153, a hallmark of ER stress, was upregulated (at 4h of exposure), followed by activation of ER-specific caspase-12 and its downstream molecule caspase-3 (at 16h of exposure). The pan caspase inhibitor, Z-VAD, and BAPTA-AM were able to reverse the Cd-induced cell death and ER stress, respectively. Furthermore, the mitochondrial membrane potential (DeltaPsi(m)) was depolarized significantly and cytochrome c was released after 24h of exposure to Cd and followed by mild activation of caspase-9 at the 36-h time point, indicating that mitochondria stress is a late event. Therefore, we concluded that ER is the major killer organelle in Cd-induced mesangial cell apoptosis and that calcium oscillation plays a pivotal role.
Asunto(s)
Apoptosis/efectos de los fármacos , Cadmio/toxicidad , Retículo Endoplásmico/efectos de los fármacos , Mesangio Glomerular/efectos de los fármacos , Animales , Línea Celular , RatonesRESUMEN
This study summarizes our most recent findings on the mechanisms underlying the cadmium-induced death of mesangial cells, which leads to nephrotoxicity. Multiple pathways participate in cadmium-induced nephrotoxicity. In the ROS-GSK-3beta autophagy pathway, cadmium induces ROS most likely from the mitochondria, and the ROS consequently activate GSK-3beta leading to autophagic cell death. In the calcium-ERK autophagy and apoptosis pathway, cadmium stimulates calcium release from the endoplasmic reticulum, which activates ERK leading to predominantly autophagic cell death and a minor level of apoptotic cell death. In the calcium-mitochondria-caspase apoptosis pathway, cadmium-induced elevation of calcium depolarizes the mitochondrial membrane potential and then activates caspase signaling leading to apoptosis. A proposed model for cadmium-induced autophagy and apoptosis leading to nephrotoxicity is summarized in Figure 1.
Asunto(s)
Cadmio/toxicidad , Células Mesangiales/efectos de los fármacos , Células Mesangiales/patología , Animales , Muerte Celular/efectos de los fármacos , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Glucógeno Sintasa Quinasa 3/metabolismo , Glucógeno Sintasa Quinasa 3 beta , Humanos , Células Mesangiales/enzimología , Modelos Biológicos , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Human intravenous immunoglobulins (IVIG) are widely used as immunomodulators because of their ability to modify the course of various immune-mediated diseases. We investigated the mechanisms responsible for the regulatory effects of IVIG on in vitro human peripheral blood mononuclear cell (PBMC) cytokine production. Pre-incubation of PBMCs with IVIG inhibited lipopolysaccharide (LPS) and phorbol 12-myristate 13-acetate (PMA)/ionomycin stimulated cytokine secretion. Pre-incubation of PBMCs with IVIG induced a significant inhibition of LPS-stimulated (IL-6) secretion (p = 0.045); the effect on tumor necrosis factor-alpha (TNF-alpha) secretion was not significant (p = 0.234). Pre-incubation of PBMCs with IVIG inhibited IL-6 secretion (p = 0.033) stimulated with anti-CD14 antibody cross-linking but had no significant effect on TNF-alpha secretion (p = 0.125). PBMC pre-incubation with anti-CD14-blocking antibody induced a significant reduction (p = 0.042) in LPS-stimulated TNF-alpha secretion in comparison with a non-significant reduction (p =0.256) noted with IVIG pre-treatment. In contrast, pre-incubation of PBMCs with anti-CD14 antibody did not induce a significant reduction in LPS-stimulated IL-6 secretion (p = 0.166) in comparison with a significant reduction (p = 0.001) induced with IVIG pre-treatment. Our data suggest that the immunoregulatory properties of IVIG may rely on several mechanisms, some of which may be independent of CD14. Our data also showed that cross-linking cell membrane-bound IVIG with anti-human kappa- and lambda-chain antibodies resulted in cytokine secretion levels similar to those elicited by LPS. In addition, intracellular DNA staining results did not support the involvement of apoptosis in the regulatory mechanisms of IVIG. This data may further our understanding of the immunoregulatory effects exerted by IVIG on the production of inflammatory-response mediators.