Asunto(s)
Antígeno B7-H1/inmunología , Mieloma Múltiple/terapia , Mieloma Múltiple/virología , Virus Oncolíticos/inmunología , Animales , Linfocitos B/inmunología , Linfocitos B/virología , Línea Celular Tumoral , Ensayos Clínicos como Asunto , Humanos , Ratones , Mieloma Múltiple/inmunología , Células Plasmáticas/inmunología , Células Plasmáticas/virologíaAsunto(s)
Azacitidina/administración & dosificación , Ciclopentanos/farmacología , Leucemia Mieloide Aguda/metabolismo , Proteínas de Neoplasias/metabolismo , Proteómica , Pirimidinas/farmacología , Línea Celular Tumoral , Ciclopentanos/administración & dosificación , Humanos , Leucemia Mieloide Aguda/patología , Pirimidinas/administración & dosificaciónRESUMEN
Activating mutation of KRas is a genetic alteration that occurs in the majority of pancreatic tumors and is therefore an ideal therapeutic target. The ability of reoviruses to preferentially replicate and induce cell death in transformed cells that express activated Ras prompted the development of a reovirus-based formulation for cancer therapy called Reolysin. We hypothesized that Reolysin exposure would trigger heavy production of viral products leading to endoplasmic reticular (ER) stress-mediated apoptosis. Here, we report that Reolysin treatment stimulated selective reovirus replication and decreased cell viability in KRas-transformed immortalized human pancreatic duct epithelial cells and pancreatic cancer cell lines. These effects were associated with increased expression of ER stress-related genes, ER swelling, cleavage of caspase-4, and splicing of XBP-1. Treatment with ER stress stimuli including tunicamycin, brefeldin A, and bortezomib (BZ) augmented the anticancer activity of Reolysin. Cotreatment with BZ and Reolysin induced the simultaneous accumulation of ubiquitinated and viral proteins, resulting in enhanced levels of ER stress and apoptosis in both in vitro and in vivo models of pancreatic cancer. Our collective results demonstrate that the abnormal protein accumulation induced by the combination of Reolysin and BZ promotes heightened ER stress and apoptosis in pancreatic cancer cells and provides the rationale for a phase I clinical trial further investigating the safety and efficacy of this novel strategy.
Asunto(s)
Apoptosis , Estrés del Retículo Endoplásmico , Virus Oncolíticos/genética , Orthoreovirus de los Mamíferos/genética , Reoviridae , Animales , Antineoplásicos/farmacología , Ácidos Borónicos/farmacología , Bortezomib , Brefeldino A/farmacología , Caspasas Iniciadoras/metabolismo , Línea Celular Tumoral , Supervivencia Celular , Terapia Combinada , Células Epiteliales/metabolismo , Células Epiteliales/virología , Femenino , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Viroterapia Oncolítica , Virus Oncolíticos/fisiología , Orthoreovirus de los Mamíferos/fisiología , Páncreas/patología , Neoplasias Pancreáticas , Proteínas Proto-Oncogénicas/biosíntesis , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas p21(ras) , Pirazinas/farmacología , Tunicamicina/farmacología , Replicación Viral , Ensayos Antitumor por Modelo de Xenoinjerto , Proteínas ras/biosíntesis , Proteínas ras/genéticaRESUMEN
Oncolytic virotherapy with reovirus has demonstrated anti-cancer activity and minimal toxicity in clinical trials, but the mechanisms underlying these effects have not been fully elucidated. Reolysin, a proprietary formulation of reovirus for cancer therapy, stimulated selective viral replication and apoptosis in multiple myeloma (MM) cells. Reolysin-mediated apoptosis was associated with an induction of endoplasmic reticular (ER) stress-related gene expression, swelling of the endoplasmic reticulum, increases in intracellular calcium levels and a strong induction of the Bcl-2 homology 3 (BH3)-only pro-apoptotic protein NOXA. Knockdown of NOXA expression by short hairpin RNA significantly reduced the pro-apoptotic effects of Reolysin. We next showed that co-administration of Reolysin and bortezomib resulted in the dual accumulation of viral and ubiquitinated proteins, which led to enhanced ER stress, NOXA induction and apoptosis. Importantly, the combination of reovirus infection and proteasomal inhibition significantly decreased tumor burden in a xenograft and syngeneic bone disease model of MM without exhibiting adverse side effects. Our study establishes ER stress stimulation and NOXA induction as novel mediators of reovirus-induced apoptosis. Furthermore, reovirus infection can be used as a promising approach to augment the anti-myeloma activity of bortezomib by promoting additional stress to the endoplasmic reticulum of MM cells.
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Antineoplásicos/uso terapéutico , Apoptosis/efectos de los fármacos , Ácidos Borónicos/uso terapéutico , Estrés del Retículo Endoplásmico , Mieloma Múltiple/tratamiento farmacológico , Mieloma Múltiple/virología , Viroterapia Oncolítica , Orthoreovirus de los Mamíferos , Pirazinas/uso terapéutico , Animales , Bortezomib , Células Cultivadas , Humanos , Ratones , Mieloma Múltiple/metabolismo , Mieloma Múltiple/patología , Trasplante de Neoplasias , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Trasplante HeterólogoRESUMEN
BACKGROUND: Upregulation of PIM kinase expression has been reported in many malignancies, suggesting that inhibition of PIM kinase activity may be an attractive therapeutic strategy. We hypothesised that inhibition of PIM kinase activity with SGI-1776, a novel small molecule inhibitor of PIM kinase activity, would reduce the viability of renal cell carcinoma (RCC) cells and enhance the activity of sunitinib. METHODS: Immunoblotting, qRT-PCR, and gene expression arrays were carried out to identify genes modulated by SGI-1776 treatment. The anticancer activity of SGI-1776 and sunitinib was determined by viability and apoptosis assays and in tumour xenografts in vivo. RESULTS: Treatment with SGI-1776 led to a decrease in phosphorylated and total c-Myc levels, which resulted in the modulation of c-Myc target genes. SGI-1776 in combination with sunitinib induced a further reduction in c-Myc levels, which was associated with enhanced anticancer activity. siRNA-mediated knockdown of c-Myc demonstrated that its expression has a key role in regulating the sensitivity to the combination of SGI-1776 and sunitinib. Importantly, the combination significantly reduced tumour burden in two RCC xenograft models compared with single-agent therapy and was very well tolerated. CONCLUSION: These data indicate that targeting PIM kinase signalling is a promising treatment strategy for RCC.
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Antineoplásicos/farmacología , Carcinoma de Células Renales/patología , Imidazoles/farmacología , Indoles/farmacología , Neoplasias Renales/patología , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas c-pim-1/antagonistas & inhibidores , Piridazinas/farmacología , Pirroles/farmacología , Animales , Carcinoma de Células Renales/enzimología , Línea Celular Tumoral , Femenino , Humanos , Inmunohistoquímica , Neoplasias Renales/enzimología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Fosforilación , Proteínas Proto-Oncogénicas c-myc/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , SunitinibRESUMEN
Heat-shock proteins (HSPs) are molecular chaperones that regulate protein folding to ensure correct conformation and translocation and to avoid protein aggregation. Heat-shock proteins are increased in many solid tumours and haematological malignancies. Many oncogenic proteins responsible for the transformation of cells to cancerous forms are client proteins of HSP90. Targeting HSP90 with chemical inhibitors would degrade these oncogenic proteins, and thus serve as useful anticancer agents. This review provides an overview of the HSP chaperone machinery and the structure and function of HSP90. We also highlight the key oncogenic proteins that are regulated by HSP90 and describe how inhibition of HSP90 could alter the activity of multiple signalling proteins, receptors and transcriptional factors implicated in carcinogenesis.
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Antineoplásicos/uso terapéutico , Sistemas de Liberación de Medicamentos/métodos , Proteínas HSP90 de Choque Térmico/antagonistas & inhibidores , Neoplasias/tratamiento farmacológico , Progresión de la Enfermedad , Proteínas HSP90 de Choque Térmico/química , Proteínas HSP90 de Choque Térmico/fisiología , Respuesta al Choque Térmico/genética , Respuesta al Choque Térmico/fisiología , Humanos , Modelos Biológicos , Relación Estructura-ActividadRESUMEN
2-(Thiocyanomethylthio)benzothiazole (TCMTB) is a biocide used in the leather, pulp and paper, and water-treatment industries. TCMTB may enter aquatic ecosystems during its manufacture and use. TCMTB is environmentally unstable; therefore, it is important to evaluate the toxicity of the more persistent degradation products. This study compared the toxicity of TCMTB with its degradation products 2-mercaptobenzothiazole (2-MBT), 2-(methylthio)benzothiazole (MTBT), benzothiazole (BT), and 2-hydroxybenzothiazole (HOBT). Toxicity was determined using Ceriodaphnia dubia 48-hour acute and 7-day chronic test protocols. TCMTB was the most toxic compound evaluated in both the acute and chronic tests with EC50s of 15.3 and 9.64 microg/L, respectively. 2-MBT, the first degradation product, was the second most toxic compound with acute and chronic EC50s of 4.19 and 1.25 mg/L, respectively. The toxicity of MTBT and HOBT were similar with acute EC50s of 12.7 and 15.1 mg/L and chronic EC50s of 6.36 and 8.31 mg/L, respectively. The least toxic compound was BT with acute and chronic EC50s of 24.6 and 54.9 mg/L, respectively. TCMTB was orders of magnitude more toxic than its degradation products. Toxicity data on these benzothiazole degradation products is important because of concerns regarding their release, degradation, persistence, and non-target organism effects in aquatic ecosystems.
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Cladóceros/efectos de los fármacos , Tiazoles/toxicidad , Tiocianatos/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Benzotiazoles , Cladóceros/fisiología , Dosificación Letal Mediana , Nivel sin Efectos Adversos Observados , Tiazoles/química , Tiocianatos/química , Pruebas de Toxicidad Aguda , Pruebas de Toxicidad CrónicaRESUMEN
B cell chronic lymphocytic leukemia (CLL) is the most prevalent adult leukemia in the Western hemisphere, yet many biological and molecular features of the disease remain undefined. CLL cells generate increased levels of radical species such as superoxide and nitric oxide (NO), which is associated with mitochondrial DNA mutations. Considering that NO levels can affect mitochondrial biogenesis, we hypothesized that the inherent nitrosative stress in CLL cells may lead to hyperactive mitochondrial biogenesis. Here we report that primary CLL cells contained significantly more mitochondria than normal lymphocytes and that their mitochondrial mass was significantly related to endogenous NO levels. Expression of the mitochondrial biogenesis factors nuclear respiratory factor-1 and mitochondrial transcription factor A was elevated in most CLL specimens examined and appeared to be related to cellular NO levels. Treatment of B cells with exogenous NO caused a substantial increase in mitochondrial mass. In vitro sensitivity of CLL cells to fludarabine was highly related to mitochondrial mass in that cells with greater mitochondrial mass were less sensitive to the drug. Taken together, our results suggest that NO is a key mediator of mitochondrial biogenesis in CLL and that modulation of mitochondrial biogenesis by NO may alter cellular sensitivity to fludarabine.