RESUMEN
We have previously shown that overexpression of RASSF1A inhibits the growth of human gastric cancer SGC7901 cells, but the underlying mechanism remains unknown. In this study, the differential protein expression by RASSF1A gene in human gastric cancer cell line SGC7901 was determined by 2-D gel electrophoresis combined with matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and bioinformatics. Differential expression analysis of the protein profiles by RASSF1A gene identified a total of 35 protein spots, of which 10 were up-regulated and 25 were down-regulated. Eight proteins were identified by MALDI-TOF MS: Galectin-1, TRP-14, ACBP, PSMB5, PSMB4, TIM, vimentin, CD79α. RASSF1A up-regulated the mRNA expression of Galectin-1, TRP-14, ABCP in SGC7901. RASSF1A also led to an increased expression of Galectin-1 protein in SGC7901 confirmed by western blotting and immunocytochemistry analysis. RASSF1A inhibited the activity of NF-κB in SGC7901 cells. These data indicated that Galectin-1 may be playing a role in RASSF1A signaling in SGC7901.
Asunto(s)
Galectina 1/metabolismo , Regulación Neoplásica de la Expresión Génica , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Proteínas Supresoras de Tumor/genética , Western Blotting , Línea Celular Tumoral , Electroforesis en Gel Bidimensional , Galectina 1/biosíntesis , Galectina 1/genética , Humanos , ARN Neoplásico/genética , ARN Neoplásico/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de Señal , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Proteínas Supresoras de Tumor/metabolismo , Regulación hacia ArribaRESUMEN
OBJECTIVE@#To investigate the effect of basic fibroblast growth factor (FGF-2)on survivin and subcellular location of Smac in human small cell lung cancer (SCLC) cell NCI-H446.@*METHODS@#Western blot was used to detect the expression of survivin protein induced by FGF-2. The release of Smac from mitochondria to cytoplasm affected by FGF-2 was observed by Western blot and immunofluorescence. Apoptosis of NCI-H446 cells was detected with flow cytometry and Hoechst 33258 staining.@*RESULTS@#The expression of survivin could be up-regulated in response to FGF-2 treatment in NCI-H446 cells, and the level of survivin expression is related to the concentration and time of FGF-2 treatment. FGF-2 could inhibit the release of Smac from the mitochondria to cytoplasm induced by serum starving. FGF-2 could inhibit the apoptosis induced by serum starving.@*CONCLUSION@#FGF-2 up-regulates the expression of survivin protein in NCI-H446 cells, and blocks the release of Smac from mitochondria cytoplasm. Survivin and Smac might play important roles in the apoptosis inhibited by FGF-2.
Asunto(s)
Humanos , Apoptosis , Proteínas Reguladoras de la Apoptosis , Citoplasma , Metabolismo , Factor 2 de Crecimiento de Fibroblastos , Farmacología , Proteínas Inhibidoras de la Apoptosis , Péptidos y Proteínas de Señalización Intracelular , Metabolismo , Neoplasias Pulmonares , Metabolismo , Patología , Proteínas Asociadas a Microtúbulos , Mitocondrias , Metabolismo , Proteínas Mitocondriales , Metabolismo , Carcinoma Pulmonar de Células Pequeñas , Metabolismo , Patología , Survivin , Células Tumorales CultivadasRESUMEN
<p><b>BACKGROUND</b>The iron catalyzed lipid peroxidation plays an important role in the autodestruction of the injured spinal cord. This study was to detect the antioxidation of melatonin against spinal cord injury (SCI) in rats.</p><p><b>METHODS</b>Sity Sprague-Dawley rats were randomly divided into four groups: group A (n = 15) for laminectomyanly, group B (n = 15) for laminectomy with SCI, group C (n = 15) for SCI and intraperitoneal injection of a bolus of 100 mg/kg melatonin, and group D (n = 15) for SCI and intraperitoneal injection of saline containing 5% ethanol. The SCI of animal model was made using modified Allen's method on T12. Six rats of each group were sacrificed 4 hours after injury, and the levels of free iron and malondialdehyde (MDA) of the involved spinal cord segments were measured by the bleomycin assay and thiobarbituric acid (TBA) separately. Functional recovery of the spinal cord was assessed by Modified Tarlov's scale and the inclined plane method at 1, 3, 7, 14, 21 days after SCI. The histologic changes of the damaged spinal cord were also examined at 7 days after SCI.</p><p><b>RESULTS</b>After SCI, the levels of free iron and MDA were increased significantly and the modified Tarlov's score and inclined plane angle decreased significantly in groups B and D. In group C, the Tarlov's score and inclined plane angle were increased significantly at 7, 14 and 21 days, with histological improvement.</p><p><b>CONCLUSION</b>Melatonin can reduce the level of lipid peroxidation and prevent damage to the spinal cord of rat.</p>
Asunto(s)
Animales , Masculino , Ratas , Antioxidantes , Usos Terapéuticos , Hierro , Peroxidación de Lípido , Malondialdehído , Melatonina , Usos Terapéuticos , Ratas Sprague-Dawley , Traumatismos de la Médula Espinal , Quimioterapia , Metabolismo , PatologíaRESUMEN
<p><b>OBJECTIVE</b>To study the effect of DPC4 gene expression on the growth of colon cancer cells and its mechanism.</p><p><b>METHODS</b>Expression plasmid pcDNA3.1-DPC4 was constructed and transfected into the colon cancer cell line SW620 by use of lipofectamine gene transfer technique. DPC4 protein expression was detected by Western blot and immunocytochemistry. The effect of DPC4 gene on the growth of SW620 cells was monitored by population doubling time (PDT) and cloning efficiency. The influence of DPC4 expression on p21WAF1 transcription was investigated by RT-PCR to detect p21WAF1 mRNA.</p><p><b>RESULTS</b>Successful expression of DPC4 protein was detected in the transfected SW620 cells. Compared with the control cells, PDT (74 h) of the DPC4 expressing cells was prolonged and the cloning efficiency (21%) decreased. In addition, the mRNA level of p21(WAF1) in DPC4 transfected cells was increased.</p><p><b>CONCLUSIONS</b>Overexpression of DPC4 gene inhibits the growth of colon cancer in vitro, and induction of p21(WAF1) expression may be an important functional aspect of DPC4.</p>
Asunto(s)
Humanos , Carcinoma , Genética , Metabolismo , Patología , Proteínas de Ciclo Celular , Genética , División Celular , Neoplasias Colorrectales , Genética , Metabolismo , Patología , Inhibidor p21 de las Quinasas Dependientes de la Ciclina , Proteínas de Unión al ADN , Genética , Técnicas de Transferencia de Gen , Genes Supresores de Tumor , Proteína Smad4 , Transactivadores , Genética , Transfección , Células Tumorales CultivadasRESUMEN
<p><b>OBJECTIVE</b>To observe the dynamic changes of free iron contents and its relationship to the changes of lipid peroxidation after experimental spinal cord injury (SCI).</p><p><b>METHODS</b>Sprague Dawley rats were randomly divided into three groups: Group A (n=6) received no operation; Group B (n=48) received only laminectomy (sham); and Group C (n=48) received both laminectomy and traumatic injury (SCI model). The SCI animal models were made by using an modified Allen's weight-drop device (50 g.cm) on T(12). Rats were sacrificed at 0.5, 1, 3, 6, 12, 24 hours after injury. The levels of free iron involved in spinal cord segments at different time points were measured by bleomycin assay. The malondialdehyde (MDA) was also measured by the thiobarbituric acid (TBA).</p><p><b>RESULTS</b>After SCI in Group C, the level of free iron showed a significant increase at 0.5 hour compared to Groups B and A, restored to the control level at 6 h; the level of MDA was increased at 0.5 hour, peaked at 3 hours, returned to the control level at 12 hours; the concentrations of free iron and lipid peroxidation in injured rats were significantly and positively correlated at 0.5-3 hours.</p><p><b>CONCLUSIONS</b>After SCI the levels of free iron are increased quickly and might be a major contributor to lipid peroxidation in injured spinal cord.</p>
Asunto(s)
Animales , Masculino , Ratas , Análisis de Varianza , Hierro , Metabolismo , Peroxidación de Lípido , Malondialdehído , Metabolismo , Distribución Aleatoria , Ratas Sprague-Dawley , Traumatismos de la Médula Espinal , MetabolismoRESUMEN
<p><b>AIM</b>To observe the effects of different periods of exercise on the iron status.</p><p><b>METHODS</b>Female rats were randomly divided into 3-, 6-, 12-month swimming exercise groups and their corresponding sedentary groups. The hematological indices of iron status and the non-heme iron (NHI) and total NHI (TNHI) of the organs were determined at the end of the desired period.</p><p><b>RESULTS</b>As compared with the corresponding sedentary groups, plasma iron and transferrin-iron saturation of three exercise groups were decreased without significant changes of blood hemoglobin and hematocrit. The NHI contents and TNHI of the liver, spleen and kidney were decreased. Although the NHI contents of the heart decreased, TNHI was not significantly changed. TNHI of the organs in both the exercised and sedentary rats were found to increase with age.</p><p><b>CONCLUSION</b>The exercise-induced low iron status with depleted iron storage is similar to the iron-deficiency status, but it could not be explained using the hypothesis of iron deficiency. Both the NHI redistribution and the maintained iron storage suggests the adaptation of low iron status to exercise. Therefore, the so-called exercise-induced iron deficiency could not exist.</p>