RESUMEN
The substantial availability of hypoxia-inducible factor 1 (HIF-1) for pathophysiological states, such as malignancies and ischemia, is primarily regulated post-translationally through the ubiquitin proteolytic system. The balance between degradation and stabilization of HIF-1α protein is determined by specific E3 ligases. In our search for new E3 ligases that might affect HIF-1α protein expression, we studied the effects of beta-transducin repeat-containing protein (ß-TrCP) on the hypoxic pathway in cancer cells. ß-TrCP is overexpressed in many tumors and regulates various cellular processes through mediating the degradation of important targets. Unexpectedly, we found that ß-TrCP overexpression increases HIF-1α protein expression level as well as HIF-1 transcriptional activity by stabilizing HIF-1α protein and preventing its ubiquitination and proteasomal degradation in prostate cancer cells. By using a proteomic approach, we succeeded in demonstrating that ß-TrCP interferes with the association between HIF-1α and HSP70/CHIP, a HIF-1α established E3 ligase complex. Whereas the E3 ligase activity of ß-TrCP is well known, antagonizing another E3 ligase is a new mechanism of action of this important E3. We suggest that destroying or suppressing ß-TrCP and thereby interrupting the HIF-1 pathway, could be valuable antitumor therapy.
Asunto(s)
Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Neoplasias de la Próstata/metabolismo , Regulación hacia Arriba/fisiología , Proteínas con Repetición de beta-Transducina/fisiología , Línea Celular Tumoral , Expresión Génica , Técnicas de Silenciamiento del Gen , Proteínas HSP70 de Choque Térmico/química , Proteínas HSP70 de Choque Térmico/metabolismo , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Masculino , Fosfotreonina/metabolismo , Proteómica , Activación Transcripcional/efectos de los fármacos , Activación Transcripcional/fisiología , Ubiquitina-Proteína Ligasas/metabolismo , Regulación hacia Arriba/efectos de los fármacos , Proteínas con Repetición de beta-Transducina/genética , Proteínas con Repetición de beta-Transducina/farmacologíaRESUMEN
Galectin-3 is a ß-galactoside-binding lectin that plays an important role in the modulation of immune responses. It has been shown to aggravate joint inflammation and destruction in experimental arthritis. We investigated the role of galectin-3 in TLR-induced cell activation in human synovial fibroblasts (SF) in order to better understand the mechanism(s) of the proinflammatory function of galectin-3 in arthritis. Galectin-3 expression in SF obtained from rheumatoid arthritis and osteoarthritis patients was inhibited by siRNA mediated gene-knockdown. Galectin-3 was also inhibited with modified citrus pectin (MCP), a polysaccharide galectin-3 ligand. Galectin-3 knockdown inhibited TLR-2, -3 and -4-induced IL-6 secretion, but not TLR-2, -3 and -4-mediated matrix metalloproteinase-3 or CC chemokine ligand-5 secretion. When the SF were stimulated with phorbol 12-myristate 13-acetate, a protein kinase C activator that bypasses the membranal receptors, galectin-3 knockdown no longer influenced IL-6 secretion. MCP reduced IL-6 levels in a dose-dependent manner. Our results indicate that galectin-3 is a positive sensor-regulator of TLR-induced IL-6 secretion in human synovial fibroblasts, thus adding new insights into the mechanisms by which galectin-3 augments synovial inflammation. These findings corroborate the potential role of glycan inhibitors of galectin-3 as a therapeutic approach for the treatment of inflammatory arthritis.
Asunto(s)
Fibroblastos/metabolismo , Galectina 3/metabolismo , Transducción de Señal , Membrana Sinovial/citología , Receptores Toll-Like/metabolismo , Quimiocina CCL5/metabolismo , Fibroblastos/efectos de los fármacos , Técnicas de Silenciamiento del Gen , Humanos , Interleucina-6/metabolismo , Lipopéptidos/farmacología , Lipopolisacáridos/farmacología , Metaloproteinasa 3 de la Matriz/metabolismo , Transducción de Señal/efectos de los fármacos , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
Breast cancer (BC) is associated with type 2 diabetes mellitus (T2DM) and obesity. Glucagon-like peptide (GLP)-1 regulates post-prandial insulin secretion, satiety, and gastric emptying. Several GLP-1 analogs have been FDA-approved for the treatment of T2DM and obesity. Moreover, GLP-1 regulates various metabolic activities across different tissues by activating metabolic signaling pathways like adenosine monophosphate (AMP) activated protein kinase (AMPK), and AKT. Rewiring metabolic pathways is a recognized hallmark of cancer, regulated by several cancer-related pathways, including AKT and AMPK. As GLP-1 regulates AKT and AMPK, we hypothesized that it alters BC cells' metabolism, thus inhibiting proliferation. The effect of the GLP-1 analogs exendin-4 (Ex4) and liraglutide on viability, AMPK signaling and metabolism of BC cell lines were assessed. Viability of BC cells was evaluated using colony formation and MTT/XTT assays. Activation of AMPK and related signaling effects were evaluated using western blot. Metabolism effects were measured for glucose, lactate and ATP. Exendin-4 and liraglutide activated AMPK in a cAMP-dependent manner. Blocking Ex4-induced activation of AMPK by inhibition of AMPK restored cell viability. Interestingly, Ex4 and liraglutide reduced the levels of glycolytic metabolites and decreased ATP production, suggesting that GLP-1 analogs impair glycolysis. Notably, inhibiting AMPK reversed the decline in ATP levels, highlighting the role of AMPK in this process. These results establish a novel signaling pathway for GLP-1 in BC cells through cAMP and AMPK modulation affecting proliferation and metabolism. This study suggests that GLP-1 analogs should be considered for diabetic patients with BC.
Asunto(s)
Neoplasias de la Mama , Exenatida , Péptido 1 Similar al Glucagón , Liraglutida , Humanos , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Exenatida/farmacología , Femenino , Liraglutida/farmacología , Péptido 1 Similar al Glucagón/metabolismo , Péptido 1 Similar al Glucagón/farmacología , Péptido 1 Similar al Glucagón/análogos & derivados , Línea Celular Tumoral , Proteínas Quinasas Activadas por AMP/metabolismo , Transducción de Señal/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Efecto Warburg en Oncología/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Ponzoñas/farmacología , Adenilato Quinasa/metabolismo , Péptidos/farmacologíaRESUMEN
BACKGROUND AND OBJECTIVE: We compared the oncologic outcomes of patients with non-muscle invasive bladder cancer (NMIBC) who underwent transurethral resection of bladder tumor (TUBRT) using sterile water vs glycine irrigation. The tumoricidal and immunogenic effects of these solutions on urothelial cancer cell lines were investigated. METHODS: The medical records of 530 consecutive patients who underwent TURBT using sterile water or glycine irrigation for NMIBC were reviewed. Recurrence and progression rates were evaluated using time dependent analyses.Bladder cancer cell lines (RT4, T24 and 5637) were treated with glycine and sterile water. Cell viability was evaluated with the XTT assay. Cell membrane calreticulin levels were evaluated with flow cytometry. Extracellular high mobility group box 1 (HMGB1) and heat shock 70 (HSP70) protein levels were evaluated using western blots. KEY FINDINGS AND LIMITATIONS: After propensity score matching each study arm comprised 161 patients. Median follow-up was 13.6 months (IQR 6.2, 24.5). The 2-year recurrence free survival was significantly lower in the sterile water vs glycine group (43% vs 71%, respectively, p<0.0001). Similarly, the 2-years progression free survival was significantly lower in the sterile water vs glycine group (85% vs 94%, respectively, p<0.014). Sterile water treatment resulted in the lowest number of viable cells. Early and late immunogenic cell death markers were markedly elevated in cells treated with glycine. CONCLUSIONS AND CLINICAL IMPLICATIONS: Sterile water compared to glycine irrigation during TURBT for NMIBC was associated with higher recurrence and progression rates. Possible explanation for these findings is the diminished immune response associated with sterile water reflected in a comparatively lesser expression of immune response inducers. PATIENT SUMMARY: We compared two irrigation fluids in non-muscle-invasive bladder cancer surgery: glycine and sterile water. Glycine outperformed sterile water in cancer recurrence, possibly boosting immunogenicity over sterile water.
RESUMEN
BACKGROUND: Physiotherapists have been urged to implement evidence-based practice for the management of low back pain (LBP). However, recent evidence suggests that many fail to do so, specifically in accordance with eliciting and addressing psychosocial factors that impact pain. The Enhanced Transtheoretical Model Intervention (ETMI) for the Treatment of patients with LBP was developed in Israel according to the clinical guidelines and demonstrated clinically and cost-effectiveness. OBJECTIVES: This study's purpose is to explore physiotherapists' perceptions of implementing evidence-based practice through the ETMI approach. DESIGN: Qualitative study. METHODS: Qualitative semi-structured interviews were conducted with 26 physiotherapists. The interviews were audio-recorded, transcribed, coded, and analyzed thematically to identify prevalent themes. RESULTS: Three main themes were identified from the transcripts, consisting of barriers and facilitators of implementing the intervention, and a meta-theme referring to shifts in the perceptions of professional role and identity. CONCLUSION: The findings suggest that physiotherapists believed that implementing ETMI and adopting a psychosocial approach to LBP can be achieved by overcoming issues around communication skills, patient reassurance, and inter-professional collaboration. Therapists also highlighted the need for physiotherapy training to gain skills in combined physical and psychological approaches.
Asunto(s)
Dolor de la Región Lumbar , Fisioterapeutas , Humanos , Dolor de la Región Lumbar/terapia , Dolor de la Región Lumbar/psicología , Fisioterapeutas/psicología , Modelo Transteórico , Actitud del Personal de Salud , Investigación Cualitativa , Práctica Clínica Basada en la EvidenciaRESUMEN
BACKGROUND: The Enhanced Transtheoretical Model Intervention (ETMI) is an approach for treating chronic low back pain (CLBP), which demonstrated clinical and cost-effectiveness outcomes. ETMI highlights reassurance, return to normal activities and encouragement of recreational physical activity. In order to optimally implement ETMI, it is important to gain an understanding of the expectations and perceptions of patients before they engage with the intervention. OBJECTIVES: To explore CLBP patients' perceptions and expectations of the ETMI method prior to their first consultation with physiotherapist. METHODS: Qualitative semi-structured interviews were conducted with 30 CLBP patients. The interviews were audio-recorded, transcribed, coded, and analysed thematically. Patients were first asked about their expectations of treatment, then they were asked to comment on the ETMI method. RESULTS: Three main themes emerged from the interviews: (1) Patient's perceptions of LBP; (2) patient's expectations from current physiotherapy and (3) Patient's expectations from ETMI method. The patients' perceptions of back pain centred on biomechanical causal factors, a desire for diagnostic tests and beliefs that rest cures the pain. Their expectations from current physiotherapy included pain reduction, passive treatment, a structured exercise program and clear information about LBP. In reference to the ETMI method, patients expected pain reduction, practical tools to self-manage, and a combination of ETMI with passive treatment. CONCLUSION: addressing issues around patient's perceptions and expectations of current physiotherapy and of the ETMI method, prior to their first consultation with physiotherapist, could be beneficial for understanding how to improve the ETMI implementation in the health care system.
Asunto(s)
Dolor de la Región Lumbar , Fisioterapeutas , Humanos , Dolor de la Región Lumbar/terapia , Motivación , Investigación Cualitativa , Modelo TransteóricoRESUMEN
This study aimed to evaluate the outcome measures and perceptions of patients with chronic low back pain (CLBP) after being treated with the Enhanced Transtheoretical Model Intervention (ETMI). In this process evaluation mixed-methods study, 30 patients with CLBP electronically completed self-reported measures (function, pain, and fear-avoidance beliefs) before and after ETMI treatment. Subsequently, each patient participated in one-on-one, semi-structured interviews, which were audio-recorded, transcribed, coded, and analyzed thematically. Quantitative analysis showed significant improvements in function (p < 0.001), pain (p < 0.001), and fear-avoidance beliefs (p < 0.001) after receiving ETMI treatment, with a large effect size (Cohen's d = 1.234). Moreover, the average number of physiotherapy sessions was 2.6 ± 0.6 for the ETMI intervention, while the annual average number in Maccabi is estimated at 4.1 ± 1.5. Three main themes emerged from the thematic analysis: (1) communication between the patient and the practitioner; (2) psychosocial treatment elements, and (3) ETMI as a long-term solution for CLBP. The findings of the current study highlight patients' perceived need for an open and sincere dialogue and for receiving reassurance and encouragement about their LBP. Notably, they had no problem with the fact that they did not receive passive treatment. Accordingly, together with the significant improvement in post-treatment outcome measures, patients perceived the ETMI method as a practical tool for self-managing their back problems in the long term.
Asunto(s)
Dolor de la Región Lumbar , Miedo/psicología , Humanos , Dolor de la Región Lumbar/psicología , Dolor de la Región Lumbar/terapia , Evaluación de Resultado en la Atención de Salud , Encuestas y Cuestionarios , Modelo TransteóricoRESUMEN
PURPOSE: Invasive mucinous adenocarcinoma (IMA) is a unique subtype of lung adenocarcinoma, characterized genomically by frequent KRAS mutations or specific gene fusions, most commonly involving NRG1. Comprehensive analysis of a large series of IMAs using broad DNA- and RNA-sequencing methods is still lacking, and it remains unclear whether molecular subtypes of IMA differ clinicopathologically. EXPERIMENTAL DESIGN: A total of 200 IMAs were analyzed by 410-gene DNA next-generation sequencing (MSK-IMPACT; n = 136) or hotspot 8-oncogene genotyping (n = 64). Driver-negative cases were further analyzed by 62-gene RNA sequencing (MSK-Fusion) and those lacking fusions were further tested by whole-exome sequencing and whole-transcriptome sequencing (WTS). RESULTS: Combined MSK-IMPACT and MSK-Fusion testing identified mutually exclusive driver alterations in 96% of IMAs, including KRAS mutations (76%), NRG1 fusions (7%), ERBB2 alterations (6%), and other less common events. In addition, WTS identified a novel NRG2 fusion (F11R-NRG2). Overall, targetable gene fusions were identified in 51% of KRAS wild-type IMAs, leading to durable responses to targeted therapy in some patients. Compared with KRAS-mutant IMAs, NRG1-rearranged tumors exhibited several more aggressive characteristics, including worse recurrence-free survival (P < 0.0001). CONCLUSIONS: This is the largest molecular study of IMAs to date, where we demonstrate the presence of a major oncogenic driver in nearly all cases. This study is the first to document more aggressive characteristics of NRG1-rearranged IMAs, ERBB2 as the third most common alteration, and a novel NRG2 fusion in these tumors. Comprehensive molecular testing of KRAS wild-type IMAs that includes fusion testing is essential, given the high prevalence of alterations with established and investigational targeted therapies in this subset.
Asunto(s)
Adenocarcinoma Mucinoso/clasificación , Adenocarcinoma Mucinoso/genética , Neoplasias Pulmonares/clasificación , Neoplasias Pulmonares/genética , Adenocarcinoma Mucinoso/patología , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Neoplasias Pulmonares/patología , Masculino , Persona de Mediana Edad , Mutación , Invasividad NeoplásicaRESUMEN
PURPOSE: External beam radiotherapy is one of the treatment options for organ-confined prostate cancer. A total dose of 70 to 81 Gray (Gy) is given daily (1.8-2.5 Gy/d), with a dose rate of 3 to 6 Gy/min over 28 to 45 treatments during 8 to 9 weeks. We applied the latest technological development in linear accelerators for enabling a wide range of dose rates (from 0.2-21 Gy/min) to test the effect of different delivery dose rates on prostate tumor growth in an animal xenograft model. MATERIALS AND METHODS: A prostate cancer xenograft model was established in CD1/nude mice by means of PC-3 and CL-1 cells. The animals were radiated by a TrueBeam linear accelerator that delivered 4 dose rates ranging from 0.6 to 14 Gy/min, and reaching a total dose of 20 Gy. The mice were weighed and monitored for tumor development twice weekly. A 2-way analysis of variance was used to compare statistical differences between the groups. RESULTS: Tumor growth was inhibited by radiation at all 4 dose rates in the 20 study mice compared to no radiation (n = 5, controls). The most significant reduction in tumor volumes was observed when the same dose of radiation was delivered at a rate of 0.6 Gy/min (P < .01). The animals' weights were not affected by any dose rate. CONCLUSIONS: Delivery of radiation with a TrueBeam linear accelerator at the lowest possible rate was most effective in prostate cancer growth inhibition and might be considered a preferential treatment mode for localized prostate cancer.
Asunto(s)
Aceleradores de Partículas , Neoplasias de la Próstata/radioterapia , Dosificación Radioterapéutica , Animales , Línea Celular Tumoral , Modelos Animales de Enfermedad , Relación Dosis-Respuesta en la Radiación , Humanos , Masculino , Ratones , Neoplasias de la Próstata/patología , Resultado del Tratamiento , Carga Tumoral , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
BACKGROUND: We evaluated whether detection of prostate-specific antigen (PSA) and human kallikrein 2 (hK2) transcripts in the peripheral blood during brachytherapy could predict biochemical outcome. METHODS: Eighty-one patients who underwent (125)Iodine-based brachytherapy for localized prostate cancer (Gleason score <8, PSA <20 ng/ml, stage Asunto(s)
Biomarcadores de Tumor/sangre
, Braquiterapia
, Antígeno Prostático Específico/sangre
, Neoplasias de la Próstata/sangre
, Neoplasias de la Próstata/radioterapia
, Calicreínas de Tejido/sangre
, Anciano
, Andrógenos/metabolismo
, Estudios de Seguimiento
, Humanos
, Masculino
, Persona de Mediana Edad
, Periodo Posoperatorio
, Valor Predictivo de las Pruebas
, Pronóstico
, Neoplasias de la Próstata/diagnóstico
, ARN Mensajero/sangre
, Resultado del Tratamiento
RESUMEN
Hypoxia-inducible factor-1 (HIF-1) is a key transcription factor in the signaling pathway that controls the hypoxic responses of cancer cells. Activation of the HIF system has been observed in carcinogenesis and numerous cancers. We found an interaction between a member of the mammalian septin gene family (MSF-A) and the HIF system. MSF-A is a nuclear protein that interacts with HIF-1alpha protein to prevent its ubiquitination and degradation, thus activating the HIF transcriptome. Cells overexpressing MSF-A protein exhibit increased HIF transcriptional activity and higher proliferation rates in vitro and in vivo. Xenograft-derived human tumors from these cells were larger and more vascular. These findings link a function of a septin protein with angiogenesis through activation of the HIF pathway.
Asunto(s)
GTP Fosfohidrolasas/fisiología , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Neovascularización Patológica , Proliferación Celular , Transformación Celular Neoplásica , Humanos , Masculino , Neoplasias de la Próstata/patología , Septinas , Transducción de Señal , Transcripción Genética , Células Tumorales CultivadasRESUMEN
In vitro and in vivo studies have shown that 1alpha,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] inhibits angiogenesis in cancer. We now examined whether the antiangiogenic effects of 1,25(OH)(2)D(3) are mediated by the hypoxia-inducible factor (HIF)-1 pathway. Our results showed that 1,25(OH)(2)D(3) reduces the protein expression of both the regulated HIF-1alpha subunit and the vascular endothelial growth factor (VEGF) in various human cancer cells. 1,25(OH)(2)D(3) also inhibited HIF-1 transcriptional activity (measured by reporter gene assay) as well as HIF-1 target genes, including VEGF, ET-1, and Glut-1. We also showed that 1,25(OH)(2)D(3) inhibits cell proliferation under hypoxia. Using HIF-1alpha knockout colon cancer cells, we show that the inhibition of the hypoxia-induced VEGF by 1,25(OH)(2)D(3) is mediated through a HIF-dependent pathway. Because HIF-1 is a major positive contributor in human tumorigenesis and angiogenesis, we believe that its inhibition by 1,25(OH)(2)D(3) strengthens the rationale to use vitamin D and its low-calcemic analogues in cancer chemoprevention and therapy.
Asunto(s)
Calcitriol/farmacología , Neoplasias del Colon/patología , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Neoplasias de la Próstata/patología , Factor A de Crecimiento Endotelial Vascular/metabolismo , Hipoxia de la Célula/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Neoplasias del Colon/genética , Neoplasias del Colon/metabolismo , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Células HCT116 , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/biosíntesis , Subunidad alfa del Factor 1 Inducible por Hipoxia/deficiencia , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Masculino , Neoplasias de la Próstata/genética , Biosíntesis de Proteínas/efectos de los fármacos , Activación Transcripcional/efectos de los fármacos , Factor A de Crecimiento Endotelial Vascular/genéticaRESUMEN
Bladder cancer (BC) is generally divided into non-muscle-invasive BC (NMIBC) and muscle-invasive BC (MIBC). The standard treatment protocol for MIBC patients is radical cystectomy preceded by neoadjuvant chemotherapy (NAC). About one-half of the MIBC patients show a priori resistance to chemotherapy, and are therefore exposed to the risks of disease progression and toxicity from ineffective NAC. The discovery of microRNA (miRNA) regulation in tumorigenesis has provided new directions for the development of a new type of BC biomarkers. In this review, we describe the emerging miRNAs as BC biomarkers for different purposes, including diagnosis, prognosis and therapeutic response. miRNA expression profile changes with alteration of the tissue phenotype. This phenomenon is utilized to predict tumor diagnosis, cancer subclass, disease stage, prognosis and therapeutic response. We classified the miRNAs which are involved in bladder cancer according to malignant potential, chemoresistance, discrimination between normal to cancerous and clinical outcome. Focusing on the major obstacle regarding MIBC patient's NAC response, we summarized the miRNAs that are deregulated and have the potential to identify the patients resistant to NAC, such as miR-34, miR-100, miR-146b and miR-9 and miR-193a-3p. In conclusion, miRNAs expression profile of bladder cancer patient is a promising tool that can serve as biomarker for different aims. Based on this profile we propose upfront radical cystectomy instead of standard NAC to those MIBC patients who are at higher risk for chemoresistance and poor response.
Asunto(s)
Biomarcadores de Tumor/genética , Manejo de la Enfermedad , Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , Neoplasias de la Vejiga Urinaria/terapia , Biomarcadores de Tumor/metabolismo , Perfilación de la Expresión Génica , Humanos , MicroARNs/metabolismo , Pronóstico , Neoplasias de la Vejiga Urinaria/genética , Neoplasias de la Vejiga Urinaria/metabolismo , Neoplasias de la Vejiga Urinaria/patologíaRESUMEN
BACKGROUND: In 1999 Cardno et al reported that long CAG repeats in the calcium-activated potassium channel gene hSKCa3/KCNN3 are associated with higher negative symptom dimension scores in schizophrenia patients. There has been no attempt to replicate the results. In this study, we investigated whether a symptom polymorphism of schizophrenia is associated with both the CAG repeat numbers and the difference in allele sizes. METHODS: We tested the association of CAG repeats with symptom models of schizophrenia in 117 unrelated Jewish patients. A multivariate analysis (MANOVA) of two models of schizophrenia with the repeat distribution and the difference in allele sizes was performed. RESULTS: We found a significant positive association of the number of CAG repeats with negative syndrome, anergia, activation, and paranoid symptoms. In addition, nonparanoid schizophrenia patients who had differences in allele sizes were characterized by earlier onset of illness. CONCLUSIONS: The study supports the hypothesis that the combined effect of long CAG repeats and the differences in allele sizes contribute to symptom expression of schizophrenia, particularly on the anergia-activation-paranoid axis.
Asunto(s)
Canales de Potasio Calcio-Activados , Canales de Potasio/genética , Esquizofrenia/genética , Repeticiones de Trinucleótidos/genética , Adolescente , Adulto , Anciano , Southern Blotting , Cartilla de ADN/genética , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Escalas de Valoración Psiquiátrica , Esquizofrenia/diagnóstico , Esquizofrenia/fisiopatología , Índice de Severidad de la Enfermedad , Canales de Potasio de Pequeña Conductancia Activados por el CalcioRESUMEN
OBJECTIVES: Several studies reported contradictory findings regarding the association of major psychosis with CAG repeats in the KCNN3 gene. We investigated the contribution of the CAG repeat at the KCNN3 gene, localized to chromosome 1q21.3, to the genetic susceptibility for schizophrenia, schizoaffective and bipolar disorders. METHODS: Analysis of the number of CAG repeats and the differences in allele length were performed for Israeli Ashkenazi Jews, non-Ashkenazi Jews, and Arabs diagnosed with major psychosis (n=181) versus matched ethnic controls (n=207). RESULTS: We found no significant difference in the number of CAG repeats between the entire sample of patients and controls. However, an analysis of the differences of allele length revealed a significantly greater number of patients with identical allele length (43.1%) when compared with normal controls (30.4%). Furthermore, an earlier age of non-paranoid schizophrenia onset was found associated with differences in allele sizes. There were no significant differences in the number of CAG repeats and the differences in allele length when subjects were grouped according to gender, ethnic origins of their parents, family history, and diagnostic groups. CONCLUSIONS: Our results support the hypothesis that a contribution of the KCNN3 gene to genetic susceptibility to major psychosis and their phenotypic polymorphism may be related to the difference of allele length rather than to the number of CAG repeats.
Asunto(s)
Canales de Potasio Calcio-Activados , Canales de Potasio/genética , Trastornos Psicóticos/genética , Repeticiones de Trinucleótidos/genética , Árabes , Secuencia de Bases , ADN/genética , Cartilla de ADN , Etnicidad , Genotipo , Humanos , Israel , Judíos , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo Genético , Canales de Potasio de Pequeña Conductancia Activados por el CalcioRESUMEN
Hypoxia-inducible factor 1α (HIF-1α) gene polymorphisms have been investigated for a possible role in mediating genetic predisposition to cancer. Our previous data show that men homozygous to C1772T polymorphism had 4-fold risk to develop prostate cancer. Therefore, we studied the effects of C1772T polymorphism on HIF-1α expression. HIF-1α mRNA expression levels were significantly higher in peripheral blood leukocytes of prostate cancer patients with the TT genotype compared with the CC genotype. Expression of C1772T HIF-1α in HIF-1α knockout cancer cells showed higher expression levels and stabilization of HIF-1α mRNA compared with the wild-type. Mutated HIF-1α protein half-life was similar to that of the wild-type. Hence, our data provide evidence that C1772T polymorphism causes activation of HIF-1α as a gain-of-function mechanism driven by stabilization of HIF-1α mRNA. These findings may also explain the increased risk of men homozygous to this mutation to develop prostate cancer.
Asunto(s)
Expresión Génica , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Polimorfismo de Nucleótido Simple , Neoplasias de la Próstata/genética , Anciano , Anciano de 80 o más Años , Hipoxia de la Célula , Regulación Neoplásica de la Expresión Génica , Genes Reporteros , Células HCT116 , Semivida , Homocigoto , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Leucocitos Mononucleares/metabolismo , Luciferasas de Luciérnaga/biosíntesis , Luciferasas de Luciérnaga/genética , Masculino , Persona de Mediana Edad , Neoplasias de la Próstata/metabolismo , Estabilidad del ARN , Elementos de RespuestaRESUMEN
OBJECTIVE: To assess the efficacy and safety of vaccination against pandemic H1N1 virus in patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), psoriatic arthritis (PsA), and ankylosing spondylitis (AS) compared with healthy controls. METHODS: The study population comprised 41 RA patients, 21 SLE patients, 17 PsA patients, 15 AS patients, and 25 healthy controls. All were vaccinated using the Novartis MF59-adjuvanted H1N1v monovalent influenza vaccine. The immunogenicity of the vaccine was assessed on day 1 and again 4 weeks later by hemagglutination inhibition assay. Geometric mean titers and seroconversion rates were calculated for each group. The safety of the vaccine was evaluated using the 28-joint Disease Activity Score (DAS28) for RA and PsA, the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI), and the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI). RESULTS: The proportion of baseline protective levels of antibodies against H1N1 was similar in all but the AS group, in which it was lower. The geometric mean titers increased significantly in all 5 groups. A substantial proportion of patients and controls responded to the vaccine. The healthy controls demonstrated a better response than each of the other groups: 84% versus 56% for RA, 67% for SLE, 59% for PsA, and 53% for AS. Multivariate logistic regression analysis identified RA and PsA as parameters of significantly lower response. The DAS28, BASDAI, and SLEDAI remained unchanged after vaccination. CONCLUSION: Vaccination against pandemic H1N1 using an adjuvanted H1N1v monovalent influenza is safe and induced an appropriate response in patients with RA, SLE, PsA, and AS.
Asunto(s)
Subtipo H1N1 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/efectos adversos , Gripe Humana/prevención & control , Enfermedades Reumáticas/inmunología , Adulto , Femenino , Humanos , Vacunas contra la Influenza/inmunología , Vacunas contra la Influenza/uso terapéutico , Masculino , Persona de Mediana Edad , Resultado del TratamientoRESUMEN
Hypoxia-inducible factor-1 (HIF-1) is a key transcription factor in the hypoxic response pathway. We recently identified a novel interaction between HIF-1alpha and the mammalian septin family member, septin 9 protein, isoform 1 (SEPT9_i1), a protein product of septin 9 transcript variant 1 (SEPT9_v1). Septins are a highly conserved family of GTP-binding cytoskeletal proteins that are implicated in multiple cellular functions, including oncogenesis. SEPT9_i1 binds and stabilizes HIF-1alpha protein and stimulates HIF-1 transcriptional activity by preventing its RACK1-mediated ubiquitination and degradation. SEPT9_i1-HIF-1 activation promotes tumor growth and angiogenesis. The effect of SEPT9_v1 silencing in prostate cancer cells was studied. SEPT9_v1 stable knockdown was generated in PC-3 cells using a specific shRNA. SEPT9_v1 silencing reduced HIF-1alpha protein expression and inhibited HIF-1 transcriptional activity. SEPT9_v1 knockdown affected cell morphology, deregulated cell cycle, and decreased migration. The antiproliferative effect of shSEPT9_v1 was abolished in HIF-1alpha knockout colon cancer cells. In vivo, SEPT9_i1 depletion reduced HIF-1alpha protein expression, cellular proliferation, tumor growth, and angiogenesis. These results provide new insights and validation for applying SEPT9_v1 as a potential target for antitumor therapy by interrupting the HIF-1 pathway.
Asunto(s)
Proteínas del Citoesqueleto/genética , Proteínas de Unión al GTP/genética , Inhibidores de Crecimiento/genética , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Neovascularización Patológica/metabolismo , Neovascularización Patológica/prevención & control , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/prevención & control , Transducción de Señal/genética , Ciclo Celular/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular , Forma de la Célula/genética , Proteínas del Citoesqueleto/deficiencia , Regulación hacia Abajo/genética , Proteínas de Unión al GTP/deficiencia , Regulación Neoplásica de la Expresión Génica/genética , Silenciador del Gen/fisiología , Inhibidores de Crecimiento/deficiencia , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/fisiología , Masculino , Neovascularización Patológica/genética , Neoplasias de la Próstata/irrigación sanguínea , Neoplasias de la Próstata/patología , Interferencia de ARN , SeptinasRESUMEN
Parathyroid hormone-related protein (PTHrP) expressed by human cancer cells enhances tumor cell growth and metastasis in vivo and it is considered as the major factor responsible for humoral hypercalcemia of malignancy. Hypoxia is a widespread feature of most solid tumors. Here, we studied the effects of hypoxia on PTHrP expression. We found that PTHrP is transcriptionally induced by prolonged (48 h) hypoxia in multiple human cancer and endothelial cell lines. Pharmacological up or downregulation of hypoxia-inducible factor (HIF) resulted in induction or reduction of PTHrP levels, respectively, implying that PTHrP hypoxic induction is mediated by HIF pathway. Analysis of PTHrP promoter revealed that both HIF-1α and HIF-2α subunits bind to specific hypoxia-responsive elements (HRE) within the P2 promoter of PTHrP. However, only HIF-2α can drive direct transcriptional activation, which can be abolished by mutation in the specific HRE. To the best of our knowledge, these results provide for the first time evidence that PTHrP is regulated by hypoxia in cancer and endothelial cells through the HIF-2 pathway. We suggest that HIF-2 induced by intratumoral hypoxia or by other genetic alterations may contribute to the pathogenesis of hypercalcemia of malignancy and cancer aggressiveness by stimulation of PTHrP expression.
Asunto(s)
Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Regulación Neoplásica de la Expresión Génica , Proteína Relacionada con la Hormona Paratiroidea/genética , Hipoxia de la Célula , Línea Celular Tumoral , Inmunoprecipitación de Cromatina , Humanos , Masculino , Proteína Relacionada con la Hormona Paratiroidea/metabolismo , Regiones Promotoras Genéticas , Neoplasias de la Próstata/metabolismo , Activación TranscripcionalRESUMEN
A critical mediator of the cellular response to hypoxia is hypoxia-inducible factor 1 (HIF-1). Increased levels of HIF-1alpha are often associated with increased tumor metastasis, therapeutic resistance, and poorer prognosis. We recently identified a novel interaction between HIF-1alpha and the mammalian septin family member, SEPT9_v1. Septins are a highly conserved family of GTP-binding cytoskeletal proteins that are implicated in multiple cellular functions, including cell division and oncogenesis. SEPT9_v1 binds and stabilizes HIF-1alpha protein and stimulates HIF-1 transcriptional activity. SEPT9_v1-HIF-1 activation promotes tumor growth and angiogenesis. The structural and functional relationships between SEPT9_v1 and HIF-1alpha were analyzed. We found that SEPT9_v1 binds specifically with HIF-1alpha but not with HIF-2alpha. The GTPase domain of SEPT9_v1 was identified as essential for HIF-1alpha binding. A GTPase domain-derived polypeptide, corresponding to amino acids 252-379, was able to disrupt HIF-1alpha-SEPT9_v1 interaction and to inhibit HIF-1 transcriptional activity. SEPT9_v1 also protected HIF-1alpha from degradation induced by HSP90 inhibition by preventing the interaction of HIF-1alpha with the RACK1 protein, which promotes its oxygen-independent proteasomal degradation. In conclusion, a new mechanism of oxygen-independent activation of HIF-1 has been identified that is mediated by SEPT9_v1 blockade of RACK1 activity on HIF-1alpha degradation.