RESUMEN
INTRODUCTION: Solitary fibrous tumour (SFT) is a rare mesenchymal tumour with intermediate malignant potential. Although this tumour arises in several sites, prostatic SFT is an extremely rare neoplasm and may prove confusing owing to the lack of clinical experience because of tumour rarity. The diagnosis may be further difficult because SFTs can manifest positive immunoreactivity for CD34 and progesterone receptor, which are known markers of prostatic stromal tumours. Herein, we describe a case of prostatic SFT that was difficult to differentiate from a prostatic stromal tumour of uncertain malignant potential because of positive immunoreactivity to CD34 and progesterone receptor. CASE REPORT: A 40-year-old Japanese man presented with lower abdominal pain. Computed tomography revealed a prostatic mass; furthermore, prostate core needle biopsy revealed proliferating bland spindle cells, without necrosis or prominent mitoses. Tumour cells were positive for CD34 and progesterone receptor on immunohistochemical analysis; thus, a prostatic stromal tumour of uncertain malignant potential was initially suspected. However, as the tumour cells showed positive immunoreactivity for STAT6, the final diagnosis was an SFT of the prostate. The patient underwent tumour resection, and at the 6-month postoperative follow-up, neither local recurrence nor distant metastasis occurred. CONCLUSION: For an accurate diagnosis of an SFT of the prostate, STAT6 immunohistochemistry should be conducted for all mesenchymal tumours of the prostate. When STAT6 immunohistochemical analysis is unfeasible, pathologists should be aware that the morphological and immunohistochemical characteristics of SFT variable from case to case and diagnose with combined analysis of several immunohistochemical markers.
Asunto(s)
Biomarcadores de Tumor/análisis , Neoplasias de la Próstata/diagnóstico , Neoplasias de la Próstata/patología , Tumores Fibrosos Solitarios/diagnóstico , Tumores Fibrosos Solitarios/patología , Adulto , Humanos , Masculino , Factor de Transcripción STAT6/biosíntesisRESUMEN
PURPOSE: To verify distress and impact thermometer (DIT) for screening emotional distress in gynecological cancer patients by Hospital Anxiety and Depression Scale total (HADS-T) as gold standard and to assess emotional changes by DIT and HADS-T. METHODS: A prospective study was conducted in newly diagnosed gynecological cancer patients during the peri-treatment period after the cancer diagnosis followed by 6-month. We defined a HADS-T score of ≥11 as being indicative of emotional distress. RESULTS: 117 patients were enrolled between May 1, 2011 and March 31, 2012, and 95 were eligible. The median age was 54 years (range 31-77). (1) From the baseline to 3-month, distress (DIT-D) ≥4 with Impact (DIT-I) ≥2 exhibited sensitivity, specificity, positive predictive value (PPV), and negative predictive values (NPV) of 0.776 [95 % confidential interval (CI) 0.688, 0.850], 0.889 (95 % CI 0.824, 0.954), 0.868 (95 % CI 0.792, 0.949), and 0.808 (95 % CI 0.731, 0.886), respectively. (2) At 6-month, DIT-D ≥2 with DIT-I ≥1 exhibited sensitivity, specificity, PPV and NPV of 0.893 (95 % CI 0.778, 1), 0.825 (95 % CI 0.707, 0.942), 0.781 (95 % CI 0.638, 0.928), and 0.917 (95 % CI 0.826, 1). (3) At 6-month, the HADS-T, DIT-D, and DIT-I scores in individual patients were significantly reduced by a mean of 4.57 (p < 0.0001), 2.34 (p < 0.0001), and 1.10 (p = 0.0031), respectively, compared with those scores of baseline (Student's paired t test), but still remained high. CONCLUSIONS: (1) On acute phase within 3-month setting, DIT; DIT-D ≥4 with DIT-I ≥2, is a reliable cut-off to screen emotional distress among gynecological cancer patients. (2) The patients' moods had improved, but not completely recovered at 6-month after the diagnosis.
Asunto(s)
Ansiedad/diagnóstico , Depresión/diagnóstico , Neoplasias de los Genitales Femeninos/psicología , Trastornos del Humor/diagnóstico , Psicometría/métodos , Adulto , Anciano , Femenino , Neoplasias de los Genitales Femeninos/terapia , Humanos , Oncología Médica , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Estudios ProspectivosRESUMEN
Ovarian and endometrial cancers diagnosed at advanced stages are often associated with malignant ascites. This study aimed to determine the safety, feasibility and efficacy of intraperitoneal (IP) docetaxel (TXT) for the treatment of ascites. A phase I study, including nine patients, was undertaken to determine the maximum tolerable dose. Efficacy was retrospectively assessed in 18 patients treated with 40-70 mg/m(2) IP TXT between 2005 and 2012. In a phase I study, the dose was safely escalated to a maximum of 70 mg/m(2), at which level no patients had grade -3 haematological adverse events. In a retrospective study of 18 patients, seven had an Eastern Cooperative Oncology Group performance status of 3; 16 had prior paclitaxel administration and two, with doses of 40 and 70 mg/m(2), experienced a serological response and a decrease in paracentesis. Thus, palliative treatment of recurrent OC should be further studied with 40 mg/m(2) among more patients, and 70 mg/m(2) could be evaluated for first-line IP chemotherapy.
Asunto(s)
Adenocarcinoma de Células Claras/tratamiento farmacológico , Antineoplásicos/administración & dosificación , Ascitis/tratamiento farmacológico , Neoplasias de los Genitales Femeninos/tratamiento farmacológico , Taxoides/administración & dosificación , Adenocarcinoma de Células Claras/complicaciones , Anciano , Ascitis/etiología , Docetaxel , Estudios de Factibilidad , Femenino , Neoplasias de los Genitales Femeninos/complicaciones , Humanos , Infusiones Parenterales , Persona de Mediana Edad , Estudios Retrospectivos , Terapia RecuperativaRESUMEN
BACKGROUND: Inhibitors of the epidermal growth factor (EGFR) signaling pathway have a major role in the treatment of KRAS wild-type colorectal cancer patients. The EGFR pathway has been shown to be activated in gastric cancer (GC). However, published data on KRAS and BRAF mutation status is limited in GC and has not been compared between GC from different geographic regions. METHODS: The prevalence of KRAS and BRAF mutations was established in 712 GC: 278 GC from the United Kingdom, 230 GC from Japan and 204 GC from Singapore. The relationship between KRAS/BRAF mutation status, DNA mismatch repair (MMR) status, clinicopathological variables and overall survival was analysed. RESULTS: Overall, 30 (4.2%) GC carried a KRAS mutation. In total, 5.8% of the UK GC, 4% of Japan GC and 1.5% of Singapore GC were KRAS mutant. KRAS mutant GC had fewer lymph node metastases in the UK cohort (P=0.005) and were more frequent in elderly patients in the Japan cohort (P=0.034). KRAS mutations were more frequent in MMR-deficient GC in the UK and the Japanese cohort (P<0.05). A BRAF mutation was only detected in a single Japanese GC. CONCLUSIONS: This large multicentre study demonstrated that KRAS mutations and DNA MMR deficiency have a role in a small subgroup of GC irrespective of country of origin, suggesting that this subgroup of GC may have developed along a common pathway. Further studies need to establish whether concomitant mutations or amplifications of other EGFR signalling pathway genes may contribute to the activation of this pathway in GC.
Asunto(s)
Reparación de la Incompatibilidad de ADN , Trastornos por Deficiencias en la Reparación del ADN , Mutación , Proteínas Proto-Oncogénicas B-raf/genética , Proteínas Proto-Oncogénicas/genética , Neoplasias Gástricas/genética , Proteínas ras/genética , Anciano , Estudios de Cohortes , Receptores ErbB/genética , Femenino , Genes ras , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Proteínas Proto-Oncogénicas p21(ras) , Neoplasias Gástricas/enzimologíaRESUMEN
BACKGROUND: Malignant pleural mesothelioma (MPM) is a rare but fatal tumour. Although most MPM patients show pleural effusion at even the early stage, it is hard to diagnose as MPM at the early stage because a sensitive and reliable diagnostic marker for MPM has not been found in plasma or pleural effusion. METHODS: In this study, we investigated whether intelectin-1 was specifically contained in MPM cells and the pleural effusion of MPM patient by immunohistochemistry, western blotting, and enzyme-linked immunosorbent assay. RESULTS: Malignant pleural mesothelioma cell lines, but not lung adenocarcinoma cell lines, secreted intelectin-1. In immunohistochemistry, epithelioid-type MPMs, but neither pleura-invading lung adenocarcinomas nor reactive mesothelial cells near the lung adenocarcinomas, were stained with anti-intelectin antibodies. Pleural effusion of MPM patients contained a higher concentration of intelectin-1 than that of lung cancer patients. CONCLUSION: These results suggest that detection of intelectin-1 may be useful for a differential diagnosis of epithelioid-type MPM in immunohistochemistry and that a high concentration of intelectin-1 in pleural effusion can be used as a new marker for clinical diagnosis of MPM.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Citocinas/metabolismo , Lectinas/metabolismo , Mesotelioma/metabolismo , Derrame Pleural Maligno/metabolismo , Neoplasias Pleurales/metabolismo , Anciano , Biomarcadores de Tumor/análisis , Línea Celular Tumoral , Citocinas/análisis , Proteínas Ligadas a GPI , Humanos , Lectinas/análisis , Masculino , Persona de Mediana Edad , Pleura/metabolismo , Derrame Pleural Maligno/químicaRESUMEN
BACKGROUND: Thromboembolic events are a major complication in ovarian cancer patients. Tissue factor (TF) is frequently overexpressed in ovarian cancer tissue and correlates with intravascular thrombosis. TF binds to coagulation factor VII (fVII), changing it to its active form, fVIIa. This leads to activation of the extrinsic coagulation cascade. fVII is produced by the liver and believed to be supplied from blood plasma at the site of coagulation. However, we recently showed that ovarian cancer cells express fVII transcripts under normoxia and that this transcription is inducible under hypoxia. These findings led us to hypothesise that ovarian cancer cells are intrinsically associated with TF-fVIIa coagulation activity, which could result in thrombosis. METHODS: In this study, we examined whether ectopically expressed fVII could cause thrombosis by means of immunohistochemistry, RT-PCR, western blotting and flow cytometry. RESULTS: Ectopic fVII expression occurs frequently in ovarian cancers, particularly in clear cell carcinoma. We further showed that ovarian cancer cells express TF-fVIIa on the cell surface under normoxia and that this procoagulant activity is enhanced by hypoxic stimuli. Moreover, we showed that ovarian cancer cells secrete microparticles (MPs) with TF-fVIIa activity. Production of this procoagulant secretion is enhanced under hypoxia. CONCLUSION: These results raise the possibility that cancer cell-derived TF-fVIIa could cause thrombotic events in ovarian cancer patients.
Asunto(s)
Factor VII/metabolismo , Neoplasias Ováricas/metabolismo , Tromboplastina/metabolismo , Tromboembolia Venosa/etiología , Hipoxia de la Célula , Línea Celular Tumoral , Micropartículas Derivadas de Células/metabolismo , Femenino , Humanos , Neoplasias Glandulares y Epiteliales/química , Neoplasias Ováricas/complicaciones , Neoplasias Ováricas/patologíaRESUMEN
The recommended therapy for genotype-2 chronic hepatitis C is a regimen of pegylated interferon alpha (peginterferon) plus ribavirin. This study was conducted to determine the value of early viral kinetics as a predictive factor for sustained virologic responder (SVR). Peginterferon alpha 2b (1.5 microg/kg/week) plus weight-based ribavirin (600-1000 mg/day) was administered to 51 patients with chronic HCV genotype 2 for 24 weeks. The HCV-RNA loads were measured at the baseline, hour 24, and week 1. The rebound index (RI, an index obtained from the viral load of week 1 divided by that of hour 24) was calculated. Compared with the baseline, the viral load at hour 24 for SVR was reduced by more than 1-log: it continued to decline thereafter, and at week 1 it was significantly lower than at hour 24 (P < 0.05). The viral load for non-SVR increased again between hour 24 and week 1. The SVR of patients with RI Asunto(s)
Antivirales/uso terapéutico
, Hepacivirus/clasificación
, Hepatitis C Crónica/tratamiento farmacológico
, Interferón-alfa/uso terapéutico
, Ribavirina/uso terapéutico
, Carga Viral
, Adulto
, Anciano
, Antivirales/administración & dosificación
, Pueblo Asiatico
, Femenino
, Genotipo
, Hepacivirus/genética
, Hepacivirus/aislamiento & purificación
, Hepatitis C Crónica/virología
, Humanos
, Interferón alfa-2
, Interferón-alfa/administración & dosificación
, Masculino
, Persona de Mediana Edad
, Polietilenglicoles
, ARN Viral/sangre
, Proteínas Recombinantes
, Ribavirina/administración & dosificación
, Resultado del Tratamiento
RESUMEN
Tissue factor (TF), the protease receptor initiating the coagulation system, functions in vascular development, angiogenesis, and tumor cell metastasis by poorly defined molecular mechanisms. We demonstrate that immobilized ligands for TF specifically support cell adhesion, migration, spreading, and intracellular signaling, which are not inhibited by RGD peptides. Two-hybrid screening identified actin-binding protein 280 (ABP-280) as ligand for the TF cytoplasmic domain. Extracellular ligation of TF is necessary for ABP-280 binding. ABP-280 recruitment to TF adhesion contacts is associated with reorganization of actin filaments, but cytoskeletal adaptor molecules typically found in integrin-mediated focal contacts are not associated with TF. Chimeric molecules of the TF cytoplasmic domain and an unrelated extracellular domain support cell spreading and migration, demonstrating that the extracellular domain of TF is not involved in the recruitment of accessory molecules that influence adhesive functions. Replacement of TF's cytoplasmic Ser residues with Asp to mimic phosphorylation enhances the interaction with ABP-280, whereas Ala mutations abolish coprecipitation of ABP-280 with immobilized TF cytoplasmic domain, and severely reduce cell spreading. The specific interaction of the TF cytoplasmic domain with ABP-280 provides a molecular pathway by which TF supports tumor cell metastasis and vascular remodeling.
Asunto(s)
Adhesión Celular/fisiología , Movimiento Celular/fisiología , Proteínas Contráctiles/metabolismo , Proteínas de Microfilamentos/metabolismo , Tromboplastina/metabolismo , Sitios de Unión , Moléculas de Adhesión Celular/metabolismo , Línea Celular Transformada , Precipitación Química , Citoplasma/metabolismo , Filaminas , Quinasa 1 de Adhesión Focal , Proteína-Tirosina Quinasas de Adhesión Focal , Humanos , Ligandos , Fosforilación , Proteínas Tirosina Quinasas/metabolismo , Células Tumorales CultivadasRESUMEN
Carboplatin is one of the most commonly used and well-tolerated agents for gynecologic malignancies. The rate of hypersensitivity reactions (HSRs) in the overall population of patients receiving carboplatin has been reported to increase after multiple doses of the agent. We retrospectively analyzed the incidence, clinical features, management, or outcome of carboplatin-related HSRs in 113 Japanese patients with gynecologic malignancies and the possibility of rechallenge with the drug. We intravenously administered carboplatin after paclitaxel or docetaxel. Mild HSRs are resolved by temporary interruption of carboplatin infusion, an additional antihistamine, and/or a corticosteroid. If HSRs arose, carboplatin was diluted, not exceeding 1 mg/mL, and slowly infused over 2 hours in subsequent cycles. Ten patients experienced carboplatin HSRs, with an overall incidence of 8.85%. The first HSR episode was mild in all cases. When retreated with carboplatin, 4 exhibited severe HSRs. More than 9 cycles and/or more than 5000 mg of carboplatin administration significantly increased the incidence of HSRs. In particular, carboplatin treatment beyond 15 cycles and/or 8000 mg increased the risk of severe HSRs (P < 0.0001). The incidence of HSRs in the ovarian carcinoma group was significantly greater than that in the uterine carcinoma group (P = 0.0046). Careful attention should be paid to HSRs during carboplatin treatment beyond 9 cycles and/or 5000 mg. The rate of severe HSRs greatly increases beyond 15 cycles and/or 8000 mg. Further studies are needed to identify potential risk factors that may contribute to the development of carboplatin HSRs and to decrease the risk of reactions.
Asunto(s)
Antineoplásicos/efectos adversos , Carboplatino/efectos adversos , Hipersensibilidad a las Drogas/etiología , Neoplasias Ováricas/tratamiento farmacológico , Neoplasias del Cuello Uterino/tratamiento farmacológico , Adenocarcinoma/tratamiento farmacológico , Adenocarcinoma/secundario , Adenocarcinoma de Células Claras/tratamiento farmacológico , Adenocarcinoma de Células Claras/secundario , Adulto , Anciano , Anciano de 80 o más Años , Cistadenocarcinoma Seroso/tratamiento farmacológico , Cistadenocarcinoma Seroso/secundario , Neoplasias Endometriales/tratamiento farmacológico , Neoplasias Endometriales/secundario , Femenino , Humanos , Incidencia , Persona de Mediana Edad , Recurrencia Local de Neoplasia/tratamiento farmacológico , Recurrencia Local de Neoplasia/patología , Neoplasias Ováricas/patología , Estudios Retrospectivos , Resultado del Tratamiento , Neoplasias del Cuello Uterino/patologíaRESUMEN
Uterine cervical and endometrial cancers are common malignant solid neoplasms for which there are no useful prognostic markers. In this study, we evaluate the relationship between ATP-binding cassette superfamily F2 (ABCF2) expression and clinical factors including clinical stage, histologic type, grade and prognosis in uterine cervical and endometrial cancer. Two hundred and sixty seven cervical and 103 endometrial cancers were studied. ATP-binding cassette superfamily F2 cytoplasmic expression was detected by immunohistochemical staining and scored as positive or negative. Among cervical cancer cases, 149 (55.8%) expressed ABCF2. The overall survival was longer in ABCF2-negative than ABCF2-positive cases (P=0.0069). Statistically significant prognostic factors for survival were ABCF2 positivity (risk ratio (rr)=1.437), old age (rr=1.550) and advanced stage (rr=2.577). ATP-binding cassette superfamily F2 positivity was an independent prognostic factor by multivariate proportional hazard test (P=0.0002). Among endometrial cancer cases, 72 (69.9%) were cytoplasmic ABCF2 positive. However, there was no significant relationship between ABCF2 expression and age, clinical stage, histologic type, histologic grade, oestrogen receptor status or prognosis. ATP-binding cassette superfamily F2 expression may be a useful prognostic marker in cervical but not endometrial cancer. The role of ABCF2 protein may differ depending on the type of cancer.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/biosíntesis , Neoplasias Uterinas/genética , Neoplasias Uterinas/metabolismo , Transportadoras de Casetes de Unión a ATP/genética , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Pronóstico , Neoplasias Uterinas/patología , Neoplasias Uterinas/terapia , Adulto JovenRESUMEN
Tissue factor (TF), the cell-surface receptor for coagulation factor VIIa, supports metastasis. Equally important for this process are (a) interactions of the TF cytoplasmic domain, which binds the mobility-enhancing actin-binding protein 280, and (b) the formation of a proteolytically active TF-VIIa complex on the tumor cell surface. In primary bladder carcinoma cells, we find that this complex localizes to the invasive edge, in proximity to tumor-infiltrating vessels that stain intensely for TF pathway inhibitor (TFPI-1), the major inhibitor of the protease activity of the complex. In culture, binding of VIIa to TF-expressing tumor cells is sufficient to allow cell adhesion, migration, and intracellular signaling on immobilized TFPI-1. Immobilized heparin, a mimic for extracellular matrix-associated proteoglycans, binds physiological concentrations of TFPI-1 in a conformation that supports TF-VIIa-dependent cell adhesion. Consistent with a functional role of TFPI-1 in complex extracellular matrices, we show that TF cooperates with integrin-mediated adhesion and migration on composite matrices that contain ligands for both integrins and the TF-VIIa complex. This study thus provides evidence for a novel mechanism of protease-supported migration that is independent of proteolytic matrix degradation but rather involves protease-dependent bridging of TF's extracellular domain to an ECM-associated inhibitor.
Asunto(s)
Carcinoma/metabolismo , Proteínas de Neoplasias , Tromboplastina/metabolismo , Neoplasias de la Vejiga Urinaria/metabolismo , Carcinoma/patología , Adhesión Celular , Movimiento Celular , Cisteína Endopeptidasas/metabolismo , Relación Dosis-Respuesta a Droga , Endopeptidasas/metabolismo , Epitelio/metabolismo , Epitelio/patología , Factor VIIa/metabolismo , Fibronectinas/farmacología , Glicoproteínas/metabolismo , Heparina/farmacología , Humanos , Inmunohistoquímica , Lipoproteínas/metabolismo , Lipoproteínas/farmacología , Proteínas Gestacionales/metabolismo , Transducción de Señal , Células Tumorales Cultivadas , Neoplasias de la Vejiga Urinaria/patologíaRESUMEN
There have been controversies whether maternal serum placental protein 5 (PP5)/tissue factor pathway inhibitor (TFPI)-2 is increased in the patients with preeclampsia and/or intrauterine growth restriction (IUGR). Here, we have estimated the serum PP5/TFPI-2 in these patients by a sandwich enzyme-linked immunosorbent assay with a newly developed monoclonal antibody, coupled with placental immunohistochemical studies of their placentae with semiquantitative scoring. Serum PP5/TFPI-2 level was significantly elevated only in the patients with preeclampsia alone (p=0.033), while PP5/TFPI-2 was detected significantly less intensely in the placentae of the same patients (p=0.035) in immunohistochemistry, as compared to Controls. A proteoglycan present on the placental villous surface, glypican-3, showed the same pattern of staining as PP5/TFPI-2, and there was a positive correlation (C.I.=0.506, p=0.004) between the immunohistochemical scores for these. Further experiments using HepG2 cells transfected with PP5/TFPI-2 suggested that glypican-3 could anchor PP5/TFPI-2 on the placental villi. A possibility that a decrease in glypican-3 in the placenta increases the outflow of PP5/TFPI-2, which in turn increases its serum level, was proposed. Preeclampsia and IUGR, often regarded to have the same pathological basis in common, showed distinct distributions of PP5/TFPI-2, which could be a clue to elucidate the pathogenesis of preeclampsia and IUGR.
Asunto(s)
Retardo del Crecimiento Fetal/metabolismo , Glicoproteínas/metabolismo , Glipicanos/metabolismo , Placenta/metabolismo , Preeclampsia/metabolismo , Adulto , Femenino , Retardo del Crecimiento Fetal/sangre , Glicoproteínas/sangre , Humanos , Inmunohistoquímica , Recién Nacido , Masculino , Preeclampsia/sangre , Embarazo , Sindecano-1/metabolismoRESUMEN
Pelizaeus-Merzbacher disease (PMD) is a rare X-linked leukodystrophy caused by proteolipid protein 1 (PLP1) gene mutations. Previous studies indicated that proteolipid proteins (PLPs) with disease-associated mutations are misfolded and trapped in the endoplasmic reticulum (ER) during transportation to the cell surface, which eventually leads to oligodendrocyte cell death in PMD. Here we report a PMD patient with a very mild phenotype carrying a novel mutation (485G-->T) in exon 4 of the PLP1 gene that causes a Trp(162)Leu substitution in the protein. We also investigated intracellular trafficking of this mutant PLP in COS-7 cells. Transiently transfected mutant PLP(W162L) fused to an enhanced green fluorescent protein (EGFP) or a short peptide tag was not carried to the plasma membrane. However, in contrast to previous studies, this mutant PLP was not retained in the ER, indicating an escape of the newly translated protein from the quality control machinery. We also found that the mutant PLP accumulated in the nuclear envelope (NE) in a time-dependent manner. This mutant PLP, with its distribution outside the ER and a very mild phenotype, supports the idea that accumulation of misfolded mutant protein in the ER causes the severe phenotype of PMD.
Asunto(s)
Proteínas de la Membrana/metabolismo , Proteína Proteolipídica de la Mielina/metabolismo , Enfermedad de Pelizaeus-Merzbacher/metabolismo , Animales , Células COS , Preescolar , Chlorocebus aethiops , Clonación Molecular/métodos , Exones , Técnica del Anticuerpo Fluorescente/métodos , Expresión Génica/genética , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Leucina/genética , Imagen por Resonancia Magnética/métodos , Masculino , Proteínas de la Membrana/genética , Mutagénesis/fisiología , Mutación/genética , Proteína Proteolipídica de la Mielina/genética , Enfermedad de Pelizaeus-Merzbacher/genética , Enfermedad de Pelizaeus-Merzbacher/patología , Transporte de Proteínas/fisiología , Receptores de Péptidos/metabolismo , Fracciones Subcelulares/metabolismo , Factores de Tiempo , Transfección/métodos , Triptófano/genéticaRESUMEN
AIMS: The aim of the present study is to clarify the level of radioactive lymph node should be biopsied after the most radioactive SN is removed. METHODS: SNB using radionuclide was performed in our hospital for 1179 primary breast cancers between April 2000 and October 2005; most (1177/1179) were performed successfully. Our criterion for harvesting SNs is to remove tissue until no radioactive site is present. The level of radioactivity and the order of removal of each lymph node were compared with pathologic results. RESULTS: More than 2 (overall average 1.9) radioactive SNs were biopsied in 686 of 1177 breasts. Cancer positive results were recorded for 142 breasts with multiple SNs. In 142 breasts, 64 showed metastasis to the most radioactive node only, 39 showed metastasis other than the most radioactive node only, and 39 showed the most radioactive node and other radioactive nodes. Moreover, if several other criteria were applied, false-positive cases were increased significantly. CONCLUSIONS: It is necessary to harvest radioactive lymph nodes other than the most radioactive. Moreover, efforts to remove every radioactive lymph node will minimize false-negative results.
Asunto(s)
Neoplasias de la Mama/patología , Ganglios Linfáticos/efectos de la radiación , Compuestos de Organotecnecio , Ácido Fítico , Radiofármacos , Renio , Biopsia del Ganglio Linfático Centinela , Compuestos de Tecnecio , Axila , Reacciones Falso Negativas , Reacciones Falso Positivas , Femenino , Humanos , Dosis de RadiaciónRESUMEN
The insulin-like growth factor I receptor (IGF-IR) plays an essential role in the establishment and maintenance of transformed phenotype, and interference with the IGF-IR pathway by antisense or dominant-negative mutants causes reversal of the transformed phenotype in many rodent and human tumor cell lines. We stably transfected an IGF-IR antisense mRNA expression plasmid into human papillomavirus (HPV)-negative C33a cell line, HPV-16-positive SiHa cell line, and HPV-18-positive HeLa S3 cell line to determine whether the IGF-IR could be a target for cervical cancer cells, especially in the presence of HPV. Approximately 30-80% down-regulation of IGF-IR expression was observed by Western blot in antisense transfected clones. There was a little inhibition in monolayer growth in all cell lines. In C33a cells, wild-type and sense clones formed 92-146 colonies in soft agar after 3 weeks; antisense clones formed <12 colonies. In SiHa cells, wild-type and sense clones formed approximately 60 colonies after 5 weeks; antisense clones formed 0-3 colonies. In HeLa S3 cells, wild-type and sense clones formed 218-291 colonies in soft agar after 2 weeks; antisense clones formed 14-160 colonies. There was a good correlation between IGF-IR down-regulation level and inhibition of transformation in soft agar. Tumorigenesis in nude mice was strongly inhibited in HeLa S3 and SiHa clones transfected with the antisense. These results indicate that down-regulation of IGF-IR by antisense RNA can reverse the transformed phenotype of human cervical cancer cells, even when harboring malignant type HPVs.
Asunto(s)
Transformación Celular Neoplásica , ARN sin Sentido/farmacología , Receptor IGF Tipo 1/antagonistas & inhibidores , Neoplasias del Cuello Uterino/terapia , Animales , Regulación hacia Abajo , Femenino , Humanos , Factor I del Crecimiento Similar a la Insulina/farmacología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Fenotipo , Receptor IGF Tipo 1/análisis , Células Tumorales Cultivadas , Neoplasias del Cuello Uterino/química , Neoplasias del Cuello Uterino/patologíaRESUMEN
We report the establishment of stable, high-level green fluorescent protein (GFP)-expressing cell lines in vitro that permit the detection and visualization of distant micrometastases when they are implanted orthotopically in nude mice. Chinese hamster ovary cells were transfected with the dicistronic expression vector containing the humanized GFP cDNA. A stable GFP-expressing clone was selected in 1.5 microM methotrexate in vitro and injected s.c. in nude mice. Stable high-level expression of GFP was maintained in the s.c. growing tumors. To use GFP expression for metastasis studies, fragments of s.c. growing tumor, which are comprised of GFP-expressing cells, were implanted by surgical orthotopic implantation in the ovary of nude mice. Subsequent micrometastases were detected in systemic organs and could be visualized by GFP fluorescence in the lung, liver, and other organs down to the single-cell level. With this fluorescent tool, we detected and visualized for the first time tumor cells at the microscopic level in fresh viable tissue in their normal host organ. Confocal microscopy further enabled us to study physiologically relevant patterns of invasion and micrometastasis.
Asunto(s)
Neoplasias Hepáticas Experimentales/metabolismo , Neoplasias Hepáticas Experimentales/patología , Proteínas Luminiscentes/biosíntesis , Animales , Antimetabolitos Antineoplásicos/farmacología , Células CHO/metabolismo , Células CHO/fisiología , Cricetinae , Resistencia a Antineoplásicos , Femenino , Proteínas Fluorescentes Verdes , Neoplasias Hepáticas Experimentales/secundario , Proteínas Luminiscentes/genética , Metotrexato/farmacología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Microscopía Confocal , Microscopía Fluorescente , Invasividad Neoplásica , TransfecciónRESUMEN
The complete nucleotide sequence of a rat genomic DNA fragment of 6.9 kbp containing the entire N-myc gene was determined. A unique structural feature of the rat N-myc gene is the presence of two polyadenylation signals in the exon 3 region resulting in formation of two poly(A)+ N-myc mRNAs of 2.9 and 2.2 kb length. Elevated expression of the 2.9 kb mRNA, which was not due to gene amplification, was observed in normal rat tissues such as brain, adrenal and testis, and in rat tumor cells such as ascites hepatoma AH130, AH70Btc and AH7974 cells, and pituitary tumor GH3 cells. In contrast, expression of 2.2 kb mRNA, for which transcription was terminated at the upstream polyadenylation site, was observed only in GH3 cells.
Asunto(s)
Genes myc/fisiología , Neoplasias/genética , Transcripción Genética , Glándulas Suprarrenales/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Encéfalo/metabolismo , Línea Celular , Clonación Molecular , Embrión de Mamíferos/metabolismo , Exones/genética , Femenino , Genes myc/genética , Riñón/metabolismo , Hígado/metabolismo , Pulmón/metabolismo , Masculino , Datos de Secuencia Molecular , Neoplasias/metabolismo , Ovario/metabolismo , Poli A , Proteínas Proto-Oncogénicas c-myc/biosíntesis , ARN , Sondas ARN , ARN Mensajero , Ratas , Mapeo Restrictivo , Testículo/metabolismo , Timo/metabolismoRESUMEN
Gene transfection experiments demonstrated that over-expression of the s-myc gene under the control of a human metallothionein promoter induced apoptosis in cells such as rat and human glioma cells. In contrast to c-Myc-mediated apoptosis requiring withdrawal of serum growth factors, s-myc expression induced apoptosis in glioma cells in the presence of 10% fetal calf serum. Whereas, s-Myc-mediated apoptosis was suppressed in proportion to the increase of bcl-2 expression as seen in c-Myc mediated apoptosis. The s-myc gene was expressed in rat embryo cells being committed to differentiate to hypertrophic chondrocytes which undergo programmed cell death. CAT assay demonstrated that in the NH2-terminal region, the s-Myc protein contains a domain structure required for expression of transactivation activity that is approximately six times higher than that of c-Myc. Therefore, these findings strongly suggest that s-Myc may play an important role in transcription regulation of a set of genes whose expression induces programmed cell death in vitro and in vivo.
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Apoptosis , Cartílago/citología , Genes myc , Proteínas Oncogénicas/fisiología , Células 3T3 , Animales , Cartílago/embriología , Cartílago/metabolismo , Regulación de la Expresión Génica , Glioma/patología , Humanos , Ratones , Proteínas Proto-Oncogénicas/fisiología , Proteínas Proto-Oncogénicas c-bcl-2 , Ratas , Factores de Transcripción/fisiología , Células Tumorales CultivadasRESUMEN
The present detailed analysis of the sequence of the extremely long (967 bp) 5'-noncoding region of a rat brain argininosuccinate lyase cDNA clone, reveals several features of interest. Multiple copies of partial and inverted (antisense) B1 repeats and multiple upstream ATG codons are present in the region, which suggests a possibility of translational control of the argininosuccinate lyase gene expression in rat brain.
Asunto(s)
Argininosuccinatoliasa/genética , Encéfalo/enzimología , ARN Mensajero/análisis , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Codón , Datos de Secuencia Molecular , Familia de Multigenes , Biosíntesis de Proteínas , RatasRESUMEN
We previously reported that s-Myc expression effectively induces programmed cell death (PCD) by apoptosis in glioma cells that express only mutated p53. To determine the molecular mechanism of s-Myc-induced PCD, we examined the correlation between transcriptional activation of s-Myc and its ability to induce PCD. Using a reporter plasmid having an upstream promoter region containing four repeats of the hexanucleotide CACGTG, we found that s-Myc can activate transcription of a reporter gene from this plasmid. Two mutated forms of s-Myc protein, s-MycCKII and s-MycmBR, were created. While s-MycCKII whose casein kinase (CK) II cognate sequence was restored in the internal acidic domain activated transcription as efficiently as wild-type s-Myc and induced PCD in glioma cells, s-MycmBR having a mutated basic region unable to bind the CACGTG motif did not. These findings suggest that transactivation activity of s-Myc through sequence-specific DNA binding may be indispensable for induction of PCD but that lack of a CK-II cognate sequence in the internal acidic domain may have little effect on these functions of s-Myc.