RESUMEN
The Assay Guidance Manual (AGM) is an eBook of best practices for the design, development, and implementation of robust assays for early drug discovery. Initiated by pharmaceutical company scientists, the manual provides guidance for designing a "testing funnel" of assays to identify genuine hits using high-throughput screening (HTS) and advancing them through preclinical development. Combined with a workshop/tutorial component, the overall goal of the AGM is to provide a valuable resource for training translational scientists.
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Bioensayo/métodos , Descubrimiento de Drogas , Geografía , Ensayos Analíticos de Alto Rendimiento , Humanos , Investigación Biomédica TraslacionalRESUMEN
Tanacetum huronense (Lake Huron tansy), which is native to the upper Midwest region of USA and Canada, was examined for the presence of anticancer compounds using an in vitro human tumor cell proliferation inhibition assay, with glioblastoma derived cell line U-87 MG. Bioassay-directed purification of the ethyl acetate extract of the aerial portion of this plant identified six active sesquiterpenoid lactones (1-6). Among these, compounds 5 and 6 are new structural analogs. One of the most abundant isolates, tanacin (4), exhibited the greatest inhibition with an IC50 value of 4.5 µg/mL.
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Antineoplásicos Fitogénicos/aislamiento & purificación , Flavonoides/aislamiento & purificación , Glioblastoma/tratamiento farmacológico , Fitoterapia , Tanacetum/química , Antineoplásicos Fitogénicos/química , Línea Celular Tumoral , Ensayos de Selección de Medicamentos Antitumorales , Flavonoides/uso terapéutico , Humanos , Lactonas/química , Lactonas/aislamiento & purificación , Extractos Vegetales/uso terapéutico , Sesquiterpenos/química , Sesquiterpenos/aislamiento & purificaciónRESUMEN
To effectively respond to viral infections, mammals rely on the innate and adaptive immune systems. Additionally, host cellular responses, such as apoptosis also play a vital role in the host defense mechanisms. To accomplish a successful replicative strategy in vivo, animal viruses have evolved a variety of molecular mechanisms that interfere with host responses. Poxviruses in particular, represents a prime example of where animal viruses encode a wide variety of proteins necessary for replication and subversion of the host's immune and single cell responses. Several proteins that inhibit host immmunomodulatory cytokines and apoptosis of infected cells have been characterized in vaccinia virus (VV). Here, we describe the identification of a protein encoded by the tanapox virus genome (142R open reading frame) that is orthologous to the B1R protein from VV. We demonstrate that like B1R, TPV142R encodes a serine threonine kinase that can phosphorylate the tumor suppressor p53 and therefore has the potential for inhibiting apoptosis of infected cells.
RESUMEN
Parthenolide is a sesquiterpene lactone that has been isolated from Tanacetum parthenium (feverfew). Parthenolide has several biological activities including the induction of apoptosis and inhibition of NF-kappaB. Because of its activities against several tumor types and because it is relatively well tolerated, in clinical trial, parthenolide is an attractive compound for the treatment of brain tumors. However, there have been no reports concerning its ability to induce apoptosis in any brain tumor cell lines. In this report we demonstrate that treatment of glioblastoma cells with parthenolide resulted in rapid apoptosis through caspase 3/7 without a suppression of NF-kappaB activity.
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Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Neoplasias Encefálicas/tratamiento farmacológico , Glioblastoma/tratamiento farmacológico , Sesquiterpenos/farmacología , Neoplasias Encefálicas/metabolismo , Caspasa 3/metabolismo , Caspasa 7/metabolismo , Línea Celular Tumoral , Glioblastoma/metabolismo , Humanos , FN-kappa B/antagonistas & inhibidoresRESUMEN
NF-kappaB is a family of transcription factors that have been shown to be elevated in a variety of tumor types and in some cases central to their survival and growth. Here we present evidence that U-87 MG and U-118 MG growth is regulated by NF-kappaB and controlled by PDGF. NF-kappaB activity was suppressed by a dominant negative mutant of the human PDGF type beta receptor and PDGF-B chain neutralizing antibodies. Creation of cell lines that had inducible expression of shRNAs directed against either c-Rel or RelA inhibited growth almost 90% indicating that NF-kappaB plays a central role in glioblastoma growth.
Asunto(s)
Astrocitoma/patología , Proliferación Celular , Glioblastoma/patología , FN-kappa B/fisiología , Astrocitoma/genética , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Progresión de la Enfermedad , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Genes rel , Glioblastoma/genética , Humanos , FN-kappa B/antagonistas & inhibidores , Factor de Crecimiento Derivado de Plaquetas/genética , Interferencia de ARN , ARN Interferente Pequeño/farmacología , Factor de Transcripción ReIA/genéticaRESUMEN
Elucidating the secondary signaling molecules that are necessary for platelet-derived growth factor (PDGF) to stimulate tumor development will be crucial to the understanding and treatment of a variety of cancers. Several lines of evidence have indicated that the transcription factor NF-kappaB plays a central role in transformation induced by Ha-ras and Bcr-abl, but nothing is known concerning its role in transformation by PDGF. Here we demonstrate that transcription from a promoter containing NF-kappaB binding sequences as well as the DNA binding activity of NF-kappaB were increased in PDGF-B-chain-transformed mouse fibroblast cells. Focus formation of PDGF-B-chain-transformed mouse fibroblasts was suppressed by treatment with acetylsalicylic acid (ASA) and salicylic acid, which are known inhibitors of NF-kappaB activation, but other nonsteroidal anti-inflammatory drugs that do not have an effect on NF-kappaB activity did not affect focus formation in these cells. Furthermore, expression of a dominant negative mutant of IkappaBalpha, pMEIkappaBalpha67CJ, and a dominant negative mutant of p65, p65DeltaC, resulted in decreased focus formation and NF-kappaB activity. Therefore, the transcription factor NF-kappaB plays a vital role in PDGF-B chain transformation of mouse fibroblast cells, and the NF-kappaB activity is sensitive to treatment with ASA.