RESUMEN
The prevalence of non-tuberculous mycobacterium (NTM) appears to be increasing. Much of the experience in the literature about this emerging organism comes from specialised units or populations such as cystic fibrosis patients. We, therefore, aim to evaluate the experience in a general respiratory population of dealing with patients with positive culture of NTM. We did a retrospective review of medical notes of general respiratory patients from whom NTM were isolated from January 2007 to July 2012. Cystic fibrosis patients were excluded. We reviewed 37 patients' (19 males, 18 females) medical records. A total of 73 positive cultures were reviewed. 28 isolates were from sputum samples alone, 34 isolates were from bronchoalveolar lavage alone and 11 isolates were from a combination of sputum and bronchoalveor lavage (11 isolates), We found that Mycobacterium avium was the most frequently isolated Mycobacterium in our laboratory with 22 (60%) patients had Mycobacterium avium in their pulmonary cultures. Interestingly, Mycobacterium gordonae and mycobacterium intracellulare were the second commonest mycobacterium (4, 11%) cultured. We noted 2 (5%), cases of Mycobacterium szulgai, 2 (5%) cases of Mycobacterium chelonae and 2 (5%) cases of Mycobacterium abscessus. There was 1(3%) case of Mycobacterium malmoense. There is prevalence of NTM in male COPD patients (7, 89%) and femal bronchiectasis (10, 77%) patients. Of our 8 COPD patients, 6 (75%) were on inhaled corticosteroids while 2 (25%) were not. 9 (24%) patients were smokers, 11 (30%) were ex-smokers, 14 (38%) were non-smokers and the smoking status of the remaining 3 (8%) was unknown. Of the 37 patients, only 6 (16%) received treatment. However, 2 patients stopped their treatment due to treatment toxicity. We concluded that the isolation of NTM is not uncommon. Defining NTM disease is difficult and deciding which patient to be treated needs careful evaluation as treatment can potentially be very toxic.
Asunto(s)
Infecciones por Mycobacterium no Tuberculosas/epidemiología , Infecciones por Mycobacterium no Tuberculosas/microbiología , Micobacterias no Tuberculosas/aislamiento & purificación , Enfermedad Pulmonar Obstructiva Crónica/microbiología , Anciano , Bronquiectasia/epidemiología , Bronquiectasia/microbiología , Líquido del Lavado Bronquioalveolar/microbiología , Femenino , Humanos , Irlanda/epidemiología , Masculino , Persona de Mediana Edad , Enfermedad Pulmonar Obstructiva Crónica/epidemiología , Estudios Retrospectivos , Esputo/microbiologíaRESUMEN
The European Respiratory Society COPD audit was a cross-sectional, multicentre study that analysed outcomes for COPD patients admitted to hospital with an exacerbation across Europe. We present the data on patients admitted to 11 Irish hospitals that participated in the audit. Among 237 patients (123 Male), the median age was 71 years and 79 (33%) patients were current smokers. 82 (35%) patients received high-flow oxygen before admission and 43 (18%) were cared for in a dedicated respiratory ward. 54 (23%) patients required ventilatory support. Median length of stay was 7 days, 98 (41%) patients were readmitted and 211 (89%) patients were alive at the 90 day follow up point. Irish patients were more likely to receive high-flow oxygen before admission, less likely to be managed in a dedicated respiratory ward and had a higher likelihood of readmission or death within 90 days than the European average.
Asunto(s)
Enfermedad Pulmonar Obstructiva Crónica/terapia , Anciano , Anciano de 80 o más Años , Estudios Transversales , Europa (Continente)/epidemiología , Femenino , Hospitalización/estadística & datos numéricos , Humanos , Irlanda/epidemiología , Tiempo de Internación/estadística & datos numéricos , Masculino , Persona de Mediana Edad , Enfermedad Pulmonar Obstructiva Crónica/epidemiología , Fumar/epidemiología , Resultado del TratamientoRESUMEN
PURPOSE: The National Lung Screening Trial (NLST) has renewed interest in low-dose computed tomography (LDCT) screening for lung cancer. Smokers may be less receptive toward LDCT screening, however, compared with never smokers. The views of patients with COPD, a particularly high-risk group, toward LDCT screening for lung cancer are currently unknown. We therefore evaluated attitudes of patients with COPD toward LDCT screening for lung cancer. METHODS: Interviews with Irish patients with COPD who satisfied NLST eligibility criteria were conducted in clinical settings using a questionnaire based on that of a comparable study of U.S. current/former smokers of unspecified disease status. RESULTS: A total of 142 subjects had a mean age of 65.09 ± 6.07 years (46.4 % were male, mean pack years 54.5 ± 33.3, mean FEV1 59.16 ± 23 %); 97.8 % had an identifiable usual source of healthcare. Compared with data from a U.S. cohort of current/former smokers, a higher proportion of Irish COPD smokers: believed that they were at risk for lung cancer (63.6 vs. 15.7 %); believed that early detection improved chances of survival (90 vs. 51.2 ); were willing to consider LDCT screening (97.9 vs. 78.6 %); were willing to pay for a LDCT scan (68.6 vs. 36.2 %); and were willing to accept treatment recommendations arising (95.7 vs. 56.2 %; p < 0.0001 for all comparisons). CONCLUSIONS: Urban Irish smokers with COPD who would be eligible for LDCT screening are almost universally in favor of being screened and treated for screening-detected lung cancers. This readily accessible high-risk population should be actively targeted in future screening programs.
Asunto(s)
Actitud Frente a la Salud , Detección Precoz del Cáncer/psicología , Neoplasias Pulmonares/diagnóstico , Aceptación de la Atención de Salud , Enfermedad Pulmonar Obstructiva Crónica/psicología , Tomografía Computarizada por Rayos X , Población Urbana , Anciano , Femenino , Humanos , Irlanda , Masculino , Persona de Mediana Edad , Dosis de Radiación , FumarRESUMEN
The transfer of apoptosis genes to tumors is one of the most promising strategies for cancer gene therapy. We have shown that massive apoptosis occurs when wild-type p53 expression is induced in glioma cells carrying a p53 gene mutation. However, adenovirus-mediated p53 gene transfer is ineffective in causing apoptosis in glioma cells that retain a wild-type p53 genotype. We evaluated the effect of E2F-1 overexpression on the growth of gliomas in vitro and in vivo. In the in vitro study, the adenovirus-mediated transfer of exogenous E2F-1 protein precipitated generalized apoptosis in gliomas. The treatment with Ad5CMV-E2F-1 of nude mice carrying subcutaneous gliomas arrested tumor growth. Our results indicate that E2F-1 has anti-glioma activity in vitro and in vivo.
Asunto(s)
Apoptosis/fisiología , Proteínas Portadoras , Glioma/genética , Factores de Transcripción/genética , Adenovirus Humanos/genética , Animales , Apoptosis/genética , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/fisiología , Muerte Celular/genética , Muerte Celular/fisiología , Supervivencia Celular/genética , Supervivencia Celular/fisiología , Inhibidor p16 de la Quinasa Dependiente de Ciclina/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/fisiología , Modelos Animales de Enfermedad , Factores de Transcripción E2F , Factor de Transcripción E2F1 , Expresión Génica/genética , Genes Supresores de Tumor , Terapia Genética , Vectores Genéticos/genética , Glioma/fisiopatología , Glioma/terapia , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Proteína Oncogénica p21(ras)/metabolismo , Proteínas Recombinantes de Fusión/genética , Proteína de Retinoblastoma/metabolismo , Proteína 1 de Unión a Retinoblastoma , Factor de Transcripción DP1 , Factores de Transcripción/fisiología , Transfección/genética , Células Tumorales CultivadasRESUMEN
A retroviral vector containing the wild-type p53 gene under control of a beta-actin promoter was produced to mediate transfer of wild-type p53 into human non-small cell lung cancers by direct injection. Nine patients whose conventional treatments failed were entered into the study. No clinically significant vector-related toxic effects were noted up to five months after treatment. In situ hybridization and DNA polymerase chain reaction showed vector-p53 sequences in posttreatment biopsies. Apoptosis (programmed cell death) was more frequent in posttreatment biopsies than in pretreatment biopsies. Tumor regression was noted in three patients, and tumor growth stabilized in three other patients.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/terapia , Técnicas de Transferencia de Gen , Genes p53 , Terapia Genética , Neoplasias Pulmonares/terapia , Retroviridae/genética , Anciano , Secuencia de Bases , Carcinoma de Pulmón de Células no Pequeñas/patología , Cartilla de ADN , Terapia Genética/efectos adversos , Terapia Genética/métodos , Vectores Genéticos/efectos adversos , Vectores Genéticos/genética , Humanos , Neoplasias Pulmonares/patología , Masculino , Persona de Mediana Edad , Datos de Secuencia MolecularRESUMEN
The potential for pharmaceuticals to produce side effects and drug interactions is well known to medical practitioners and the lay public alike. However, the potential for alternative medicines to produce such effects is less widely known. We describe a potentially dangerous interaction between a herbal medicine and concomitant selective serotonin re-uptake inhibitor (SSRI) ingestion.
Asunto(s)
Citalopram/efectos adversos , Interacciones de Hierba-Droga , Fitoterapia , Rhodiola/efectos adversos , Inhibidores Selectivos de la Recaptación de Serotonina/efectos adversos , Adulto , Atención/efectos de los fármacos , Femenino , HumanosRESUMEN
A consistent, if not invariant, feature of cancer cells is the acquired ability to evade apoptosis. The pioneering work of Dr. Stan Korsmeyer was invaluable in characterizing the molecular foundations of cell death signaling mechanisms during normal development and during multistep carcinogenesis. This foundation now forms the basis for the rational design of therapeutic strategies to selectively activate cell death in cancer cell populations. These strategies are currently being evaluated in an increasing number of clinical trials targeting diverse tumor types.
Asunto(s)
Carcinoma/metabolismo , Carcinoma/patología , Animales , Carcinoma/genética , Carcinoma/terapia , Muerte Celular , Transformación Celular Neoplásica/patología , Progresión de la Enfermedad , Humanos , Proteínas Oncogénicas/genética , Proteínas Oncogénicas/metabolismo , Transducción de SeñalRESUMEN
We characterized the basis for the follicular lymphoproliferation in transgenic mice bearing a Bcl-2-immunoglobulin (Bcl-2-Ig) minigene representing the t(14;18) of human follicular lymphoma. Discriminatory S1 nuclease protection assays revealed that the Bcl-2-Ig transgene was overexpressed relative to endogenous mouse Bcl-2 in spleen and thymus. Western (immunoblot) analysis demonstrated the overproduction of the human 25-kilodalton Bcl-2 protein, which arose from the transgene, in spleen, thymus, and the expanded B-cell subset. Despite the generalized lymphoid pattern of deregulation, two-color flow cytometry and density gradient centrifugation indicated that the expanded lymphocytes were predominantly small, resting B cells coexpressing B220, immunoglobulin M (IgM), IgD, Ia, and kappa. Cell cycle analysis confirmed that about 97% of these expanded B cells reside in G0/G1. An extensive characterization of transgenic lines revealed a fourfold excess of IgM-IgD-expressing B cells in spleen and dramatically increased numbers in bone marrow. While resting, these cells proliferated in response to lipopolysaccharide and anti-IgM and demonstrated normal B-cell colony formation in soft agar. Moreover, these B cells, which demonstrated an extended survival in vitro even in the absence of stroma, were also resting in G0, yet were capable of proliferative responses. These findings provide consistent evidence that the accumulation of B cells after Bcl-2 overproduction is secondary to prolonged cell survival and not increased cell cycling. This suggests a unique role for Bcl-2 as a proto-oncogene that enhances cell survival independent of promoting cell division.
Asunto(s)
Linfocitos B/fisiología , Genes de Inmunoglobulinas , Inmunoglobulina D/genética , Inmunoglobulina M/genética , Proteínas Proto-Oncogénicas/genética , Animales , Antígenos de Diferenciación de Linfocitos B/análisis , Western Blotting , Ciclo Celular , Regulación de la Expresión Génica , Lipopolisacáridos/farmacología , Tejido Linfoide/fisiología , Ratones , Ratones Transgénicos , Peso Molecular , Proto-Oncogenes Mas , Proteínas Proto-Oncogénicas c-bcl-2 , ARN Mensajero/genéticaRESUMEN
BACKGROUND: Alterations of the p53 (also called TP53) gene are one of the most common abnormalities in gliomas. We have previously reported that restoration of wild-type p53 protein function in glioma cells results in programmed cell death (apoptosis). Since p53 functions are mediated by genes that directly control the tumor suppressor effect of the p53 protein, understanding the relationship between p53 and p53-related genes in glioma cells will aid in the design of more rational treatment strategies for brain tumors. PURPOSE: We conducted this study to examine the timing of the p53-mediated events preceding apoptosis. More specifically, we undertook this work to characterize the genetic and cell cycle-related factors that may increase the resistance of glioma cells to p53-induced apoptosis. METHODS: Two human glioma cell lines (U-251 MG and U-373 MG) that express mutant p53 protein and two (U-87 MG and EFC-2) that express wild-type p53 protein were used. Replication-deficient adenovirus was utilized as an expression vector to transfer exogenous p53 and p21 complementary DNAs into the glioma cells; control cells were infected with the viral expression vector alone. To monitor gene transfer and the expression of exogenous genes (as well as the expression of endogenous genes), we used western blot analyses and immunohistochemistry analyses. Flow cytometry studies of cellular DNA content were performed to determine the cell cycle phenotype of the glioma cells before and after treatment. RESULTS: p53-mediated apoptosis was preceded by elevation in the levels of the p21 (cell cycle-related) and Bax (apoptosis-related) proteins. In addition, cell cycle analyses showed that glioma cells were arrested in the G2 phase before undergoing cell death. Transfer of p21 induced a G2 block but did not induce apoptosis. Moreover, coexpression of p21 and p53 prevented glioma cells from undergoing apoptosis. Expression of exogenous p53 in wild-type p53 cells did not induce elevation of Bax levels, arrest in G2 phase, or apoptosis. CONCLUSIONS AND IMPLICATIONS: Our data confirmed the ability of wild-type p53 to induce apoptosis in p53 mutant glioma cells. In addition, our results document that p21 plays a role in protecting cells from p53-mediated programmed cell death and suggest that p53-mediated apoptosis and p21 induction may represent, at least in certain cases, opposite signals. Finally, our data suggest that over expression of p21 in gliomas may be related to resistance to treatments that induce apoptosis.
Asunto(s)
Neoplasias Encefálicas/fisiopatología , Ciclinas/fisiología , Regulación Neoplásica de la Expresión Génica , Glioblastoma/fisiopatología , Proteína p53 Supresora de Tumor/fisiología , Adenoviridae , Apoptosis , Western Blotting , Neoplasias Encefálicas/genética , Inhibidor p21 de las Quinasas Dependientes de la Ciclina , Ciclinas/biosíntesis , ADN de Neoplasias/genética , Citometría de Flujo , Vectores Genéticos , Glioblastoma/genética , Humanos , Fenotipo , Proteína p53 Supresora de Tumor/biosíntesis , Regulación hacia ArribaAsunto(s)
Calidad de Vida , Síndrome de Dificultad Respiratoria/fisiopatología , Síndrome de Dificultad Respiratoria/terapia , Adulto , Femenino , Humanos , Masculino , Modalidades de Fisioterapia , Recuperación de la Función , Síndrome de Dificultad Respiratoria/epidemiología , Síndrome de Dificultad Respiratoria/etiología , Tasa de SupervivenciaRESUMEN
The Wilms' tumor gene WT1 functions as a tumor suppressor gene, repressing transcription of several growth factors and growth factor receptors. The bcl-2 and c-myc proto-oncogenes are essential for regulation of apoptosis and cell proliferation with roles in development and oncogenesis. We found that WT1 can repress transcription of both the bcl-2 and c-myc promoters. This suggests that WT1 regulates bcl-2 and c-myc during renal development, and the loss of functional WT1 results in deregulation of bcl-2 and c-myc, contributing to tumor formation.
Asunto(s)
Proteínas de Unión al ADN/genética , Regulación de la Expresión Génica , Genes Supresores de Tumor , Genes myc , Neoplasias Renales/genética , Proteínas Proto-Oncogénicas/genética , Proto-Oncogenes , Factores de Transcripción/genética , Tumor de Wilms/genética , Secuencia de Aminoácidos , Secuencia de Bases , Humanos , Datos de Secuencia Molecular , Regiones Promotoras Genéticas , Proteínas Proto-Oncogénicas c-bcl-2 , Proteínas WT1RESUMEN
Cancer gene therapy strategies for inducing apoptosis in solid tumors may allow contemporary medicine to reassess its management of these cancers. We demonstrated previously that overexpression of the wild-type p53 gene in squamous cell carcinoma of the head and neck cell lines via adenovirus-mediated gene transfer suppressed growth both in vitro and in vivo. Here, we characterize the mechanism of the growth suppression by the exogenous p53 gene as a consequence of programmed cell death (apoptosis). One of the cell lines used in this study, Tu-138, harbors a mutated p53 gene, whereas the other cell line, MDA 686LN, possesses a wild-type p53 gene. DNA fragmentation was detected by electrophoresis in both cell lines after infection with the wild-type p53 adenovirus, Ad5CMV-p53. With the use of the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling method, 4.4% of the remaining viable Tu-138 cell population was identified as apoptotic as early as 15 h after inoculation with Ad5CMV-p53. The percentage of apoptotic cells increased to 31% at 22 h. In contrast, only 10% of the viable MDA 686LN cells (wt-p53) had undergone apoptosis 30 h after Ad5CMV-p53 infection, although the percentage of apoptotic cells rapidly increased to 60% at 48 h after infection. For in vivo analysis of apoptosis, nude mice in which squamous cell carcinoma of the head and neck cell lines had been implanted s.c. had exogenous wt-p53 transiently introduced to the tumor cells via Ad5CMV-p53 2 days later. In situ end labeling clearly illustrated apoptosis in the tumor cells. These results suggest that wt-p53 plays an important role in the induction of apoptosis in human head and neck cancer cell lines and that selective induction of apoptosis in cancer cells can be further explored as a strategy for cancer gene therapy.
Asunto(s)
Adenoviridae/genética , Apoptosis/fisiología , Carcinoma de Células Escamosas/patología , Carcinoma de Células Escamosas/terapia , Técnicas de Transferencia de Gen , Genes Virales , Genes p53 , Neoplasias de Cabeza y Cuello/patología , Neoplasias de Cabeza y Cuello/terapia , Animales , Carcinoma de Células Escamosas/genética , División Celular/fisiología , ADN Nucleotidilexotransferasa/análisis , ADN de Neoplasias/análisis , ADN de Neoplasias/metabolismo , ADN Viral/análisis , ADN Viral/metabolismo , Femenino , Terapia Genética , Neoplasias de Cabeza y Cuello/genética , Humanos , Ratones , Ratones Desnudos , Mutación , NecrosisRESUMEN
The significance of apoptosis in relation to the development and progression of prostate cancer remains largely undefined. bcl-2 is an oncogene that functions by overriding apoptosis. bcl-2 expression was localized to the basal epithelial cells in the normal human prostate with the use of immunohistochemistry. Androgen-dependent and androgen-independent prostate carcinomas were evaluated immunohistochemically for bcl-2 expression. bcl-2 was undetectable in 13 of 19 cases of androgen-dependent cancers. In contrast, androgen-independent cancers displayed diffuse, high levels of bcl-2 staining (P < 0.01). In rats, steady-state levels of bcl-2 mRNA, assessed by S1 assays, reached maximum levels 10 days following castration. Addition of exogenous testosterone with, or without, flutamide demonstrated that the increased bcl-2 mRNA resulted from androgen ablation. Our findings indicate that bcl-2 expression is augmented following androgen ablation and is correlated with the progression of prostate cancer from androgen dependence to androgen independence.
Asunto(s)
Expresión Génica , Próstata/metabolismo , Neoplasias de la Próstata/etiología , Proteínas Proto-Oncogénicas/genética , Proto-Oncogenes , Andrógenos/farmacología , Animales , Apoptosis , Masculino , Neoplasias Hormono-Dependientes/etiología , Neoplasias de la Próstata/genética , Proteínas Proto-Oncogénicas c-bcl-2 , Ratas , Ratas Sprague-DawleyRESUMEN
One of the most promising strategies in cancer gene therapy is adenoviral transfer of proapoptotic genes. We therefore evaluated the novel strategy of adenovirally overexpressing the proapoptotic Bak gene. Our results showed marked apoptosis in cancer cells in vivo and in vitro after Bak gene transfer via a binary adenoviral vector system. This effect was not seen in a caspase 3-defective cell line (MCF-7) and was abrogated in Bak-sensitive tumors after administration of the caspase inhibitor z-DEVD-fmk. Our results suggest that adenoviral-mediated overexpression of Bak provides a novel therapeutic strategy for cancer therapy, but this process appears to be caspase dependent.
Asunto(s)
Adenoviridae/genética , Caspasas/fisiología , Terapia Genética , Proteínas de la Membrana/genética , Neoplasias/terapia , Animales , Apoptosis , Femenino , Técnicas de Transferencia de Gen , Humanos , Ratones , Ratones Desnudos , Proteínas Proto-Oncogénicas c-bcl-2/fisiología , Células Tumorales Cultivadas , Proteína Destructora del Antagonista Homólogo bcl-2RESUMEN
Understanding the functional roles of the molecular alterations that are involved in the oncogenesis of prostate cancer, the second most frequent cause of cancer-related deaths among men in the United States is the focus of numerous investigations. To examine the possible significance of alterations associated with the tumor suppressor gene, MMAC/PTEN, in prostate carcinoma, the biological and biochemical effects of MMAC/PTEN expression were examined in LNCaP cells, which are devoid of a functional gene product. Acute expression of MMAC/PTEN via an adenoviral construct resulted in a dose-dependent and specific inhibition of Akt/PKB activation, consistent with the phosphatidylinositol phosphatase activity of MMAC/PTEN. MMAC/PTEN expression induced apoptosis in LNCaP cells, although to a lesser extent than that observed with p53 via an adenoviral construct. However, MMAC/PTEN expression produced a growth inhibition that was significantly greater than that achieved with p53. Overexpression of Bcl-2 in LNCaP cells blocked MMAC/PTEN- and p53-induced apoptosis but not the growth-suppressive effects of MMAC/ PTEN, suggesting that the growth regulatory effects of MMAC/PTEN involve multiple pathways. These studies further implicate the loss of MMAC/PTEN as a significant event in prostate cancer and suggest that reintroduction of MMAC/PTEN into deficient prostate cancer cells may have therapeutic implications.
Asunto(s)
Apoptosis/fisiología , Proteínas de Arabidopsis , Carcinoma/metabolismo , División Celular/fisiología , Monoéster Fosfórico Hidrolasas/fisiología , Proteínas de Plantas/metabolismo , Canales de Potasio/metabolismo , Neoplasias de la Próstata/metabolismo , Proteínas Supresoras de Tumor , Carcinoma/genética , Carcinoma/patología , Vectores Genéticos , Humanos , Masculino , Fosfohidrolasa PTEN , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/patología , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
Wild-type p53 is involved in several aspects of cell cycle control and suppression of transformation, inducing either apoptosis or G1 block in cell cycle progression. Using a recombinant adenovirus containing the wild-type p53 cDNA, the biological effects of the newly expressed wild-type p53 protein were examined in six human glioma cell lines. Three cell lines (U-251 MG, U-373 MG, and A-172) expressed endogenous mutant p53, and the other three (U-87 MG, EFC-2, and D54 MG) expressed wild-type p53. The restoration of normal p53-encoded protein in the mutant cell lines induced apoptosis as assessed by morphological studies using nuclear staining, electron microscopy, and flow cytometric assays. In wild-type p53 cell lines, however, the overexpression of wild-type p53 did not result in apoptosis but inhibited cellular proliferation rather drastically and modified the neoplastic phenotype. Differential effects suggest two pathways for glioma oncogenesis and a possible therapeutic strategy.
Asunto(s)
Adenovirus Humanos , Apoptosis , Genes p53 , Transfección/métodos , Proteína p53 Supresora de Tumor/biosíntesis , Western Blotting , División Celular , Línea Celular , Citometría de Flujo , Expresión Génica , Vectores Genéticos , Glioma/genética , Homocigoto , Humanos , Cinética , Proteínas Recombinantes/biosíntesis , Factores de Tiempo , Células Tumorales CultivadasRESUMEN
Strong evidence exists to support the tenet that activation of E2F transcription factors, via alterations in the p16-cyclin D-Rb pathway, is a key event in the malignant progression of most human malignant gliomas. The oncogenic ability of E2F has been related to the E2F-mediated up-regulation of several proteins that positively regulate cell proliferation. However, E2F may indirectly enhance proliferation by activating antiapoptotic molecules. In this work, we sought to ascertain whether E2F-1-mediated events involve the up-regulation of the antiapoptotic molecule Bcl-2. Western blot analyses showed up-regulation of Bcl-2 but not of Bcl-x(L) by 24 h after the transfer of E2F-1. Northern blot studies showed that transfer of E2F-1 also up-regulated Bcl-2 RNA. In support of these findings and the concept that E2F-1 has a direct effect in the induction of Bcl-2, we found a putative E2F binding site within the Bcl-2 sequence. Subsequent gel-mobility shift and supershift experiments involving the CTCCGCGC site in the bcl-2 promoter showed that E2F-1 bound Bcl-2. Transactivation experiments consistently showed that ectopic E2F-1 activated responsive elements located in the -1448/-1441 region in the P1 promoter region of the bcl-2 gene. As expected, other members of the E2F family of transcription factors such as E2F-2 and E2F-4 also transactivated the bcl-2 promoter. Our results demonstrate that E2F-1 modulates the expression of the antiapoptotic molecule Bcl-2 and suggest that up-regulation of Bcl-2 may favor the oncogenic role of E2F-1 and other members of the E2F family of transcription factors.
Asunto(s)
Proteínas de Ciclo Celular , Proteínas de Unión al ADN , Glioma/genética , Proteínas Proto-Oncogénicas c-bcl-2/genética , Factores de Transcripción/genética , Activación Transcripcional , Sitios de Unión , Ciclo Celular/fisiología , Factores de Transcripción E2F , Factor de Transcripción E2F1 , Factor de Transcripción E2F2 , Factor de Transcripción E2F4 , Regulación Neoplásica de la Expresión Génica , Técnicas de Transferencia de Gen , Genes bcl-2/genética , Glioma/metabolismo , Humanos , Regiones Promotoras Genéticas , Proteínas Proto-Oncogénicas c-bcl-2/biosíntesis , Factores de Transcripción/biosíntesis , Regulación hacia ArribaRESUMEN
Deregulation of c-myc, frequently implicated in oncogenesis, is associated with increased cell proliferation and also cell death. Similarly, the p53 tumor suppressor gene commonly mutated in human tumors, is known to induce apoptosis or cell cycle arrest in its wild-type conformation. Genetically altered mice simultaneously overexpressing c-myc and possessing a disrupted p53 gene were used to investigate whether c-myc mediated apoptosis requires wild-type p53. The accelerated development of malignant lymphomas in these mice was found to be a consequence of enhanced proliferation and not reduced apoptosis resulting from the synergistic effect of c-myc overexpression and p53 inactivation.
Asunto(s)
Apoptosis/genética , Genes myc , Genes p53 , Linfoma/genética , Alelos , Animales , Secuencia de Bases , División Celular/genética , Deleción Cromosómica , Cartilla de ADN , Heterocigoto , Linfoma/patología , Ratones , Ratones Transgénicos , Datos de Secuencia MolecularRESUMEN
The mechanism by which Bcl-2 oncogene expression inhibits radiation-induced apoptosis has been investigated in two mouse lymphoma cell lines: line LY-as is radiation sensitive, displays substantial radiaton-induced apoptosis, and expresses low levels of Bcl-2; line LY-ar is radiation-resistant, displays a low apoptosis propensity, and expresses 30-fold higher amount of Bcl-2 protein than does the sensitive line. We observed that upon incubation in cystine/methionine-free (C/M-) medium, radiation-induced apoptosis in the LY-ar cells was restored to levels comparable to that seen in the LY-as cells. lntracellular glutathione (GSH) concentrations in LY-ar cells incubated in C/M- medium plummeted to 50% of control values within 2 h. LY-ar cells treated with diethyl maleate (DEM) or diamide, agents that deplete cellular thiols, had increased susceptibility to radiation-induced apoptosis in a manner similar to C/M- medium. These results are consistent with the general idea that Bcl-2 expression blocks apoptosis through an antioxidant pathway that involves cellular thiols. That Bcl-2-expressing tumor cells can be sensitized by exogeneous agents that modify cellular thiols offers strategies for overcoming such resistance.
Asunto(s)
Apoptosis/fisiología , Genes bcl-2 , Proteínas Proto-Oncogénicas c-bcl-2/biosíntesis , Tolerancia a Radiación , Animales , Apoptosis/efectos de la radiación , Butionina Sulfoximina/farmacología , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Fragmentación del ADN , Relación Dosis-Respuesta en la Radiación , Glutatión/metabolismo , Cinética , Linfoma , Ratones , Proteínas Proto-Oncogénicas/biosíntesis , Factores de Tiempo , Células Tumorales Cultivadas , Proteína p53 Supresora de Tumor/biosíntesis , Proteína X Asociada a bcl-2RESUMEN
The ATP/ubiquitin-dependent 26S proteasome is a central regulator of cell cycle progression and stress responses. While investigating the application of peptide aldehyde proteasome inhibitors to block signal-induced IkappaBalpha degradation in human LNCaP prostate carcinoma cells, we observed that persistent inhibition of proteasomal activity signals a potent cell death program. Biochemically, this program included substantial upregulation of PAR-4 (prostate apoptosis response-4), a putative pro-apoptotic effector protein and stabilization of c-jun protein, a potent pro-death effector in certain cells. We also observed modest downregulation of bcl-XL, a pro-survival effector protein. However, in contrast to some recent reports stable, high level, expression of functional bcl-2 protein in prostate carcinoma cells failed to signal protection against cell death induction by proteasome inhibitors. Also in disagreement to a recent report, no evidence was found for activation of the JNK stress kinase pathway. A role for p53, a protein regulated by the proteasome pathway, was ruled out, since comparable cell death induction by proteasome inhibitors occurred in PC-3 cells that do not express functional p53 protein. These data signify that the ubiquitin/proteasome pathway represents a potential therapeutic target for prostate cancers irrespective of bcl-2 expression or p53 mutations.