RESUMEN
Local coagulation activation has been shown to impact both primary tumor growth and metastasis in mice. It is well known that components of the blood clotting cascade such as tissue factor and thrombin play a role in tumor progression by activating cellular receptors and local formation of fibrin. However, whether venous thromboembolism (VTE) or a hypercoagulable state has a direct impact on cancer progression is unknown. Here we have combined an orthotopic murine breast cancer model, using female Nod-SCID mice, with siRNA-mediated silencing of antithrombin (siAT) leading to the induction of a systemic hypercoagulable state. We show that, compared to control siRNA-treated (not experiencing a hypercoagulable state) tumor-bearing mice, siAT treated tumor-bearing mice do not show enhanced tumor growth nor enhanced metastasis. We conclude that, in this murine model for hypercoagulability, induction of a hypercoagulable state does not contribute to breast cancer progression.
Asunto(s)
Neoplasias de la Mama , Trombofilia , Humanos , Femenino , Animales , Ratones , Antitrombinas , Modelos Animales de Enfermedad , Xenoinjertos , Ratones SCID , Trombofilia/genética , Anticoagulantes , Neoplasias de la Mama/complicaciones , Neoplasias de la Mama/genética , Antitrombina III/genética , ARN Interferente PequeñoRESUMEN
Accumulating evidence suggests that a subpopulation of breast cancer cells, referred to as cancer stem cells (CSCs), have the ability to propagate a tumor and potentially seed new metastases. Furthermore, stimulation of an epithelial-to-mesenchymal transition by factors like transforming growth factor-ß (TGFß) is accompanied with the generation of breast CSCs. Previous observations indicated that bone morphogenetic protein-7 (BMP7) antagonizes the protumorigenic and prometastatic actions of TGFß, but whether BMP7 action is mechanistically linked to breast CSCs has remained elusive. Here, we have studied the effects of BMP7, BMP2 and a BMP2/7 heterodimer on the formation of human breast CSCs (ALDH(hi)/CD44(hi)/CD24(-/low)) and bone metastases formation in a preclinical model of intra-cardiac injection of MDA-MB-231 cells in athymic nude (Balb/c nu/nu) mice. The BMP2/7 heterodimer was the most efficient stimulator of BMP signaling and very effectively reduced TGFß-driven Smad signaling and cancer cell invasiveness. The tested BMPs-particularly the heterodimeric BMP2/7-strongly reduced the size of the ALDH(hi)/CD44(hi)/CD24(-/low) CSC subpopulation. In keeping with these in vitro observations, pretreatment of cancer cells with BMPs for 72 h prior to systemic inoculation of the cancer cells inhibited the formation of bone metastases. Collectively, our data support the notion that breast CSCs are involved in bone metastasis formation and describe heterodimeric BMP2/7 as a powerful TGFß antagonist with anti-metastatic potency.
Asunto(s)
Proteína Morfogenética Ósea 2/metabolismo , Proteína Morfogenética Ósea 7/metabolismo , Neoplasias Óseas/secundario , Neoplasias de la Mama/metabolismo , Células Madre Neoplásicas/fisiología , Animales , Proteínas Morfogenéticas Óseas/metabolismo , Proteínas Morfogenéticas Óseas/farmacología , Neoplasias Óseas/metabolismo , Neoplasias de la Mama/patología , Neoplasias de la Mama/prevención & control , Línea Celular Tumoral , Movimiento Celular , Humanos , Ratones , Ratones Desnudos , Células Madre Neoplásicas/efectos de los fármacos , Transducción de Señal , Proteínas Smad/genética , Transfección , Trasplante HeterólogoRESUMEN
Cancer is a major leading cause of death in the western world (following heart diseases). It poses an enormous burden on patients and society with a major impact on healthcare and economy. Once cancers have spread to the skeleton, treatment options are predominantly limited to palliation, treatment of hypercalcemia and prevention of pathological fractures. Despite the elaborate efforts of modern medicine to improve treatment, novel therapies for the treatment of solid tumors in patients with advanced disease, including metastatic bone disease, have generally failed to improve patient overall survival. Despite initial beneficial responses on metastatic tumor burden this is frequently followed by re-growth of therapy resistant, malignant metastatic bone lesions. Cancer relapse in bone coincides with devastating consequences and causes considerable morbidity. Bisphosphonates represent the current gold standard in bone metastasis therapies. Because of the progress made in our understanding of the pathogenesis of skeletal metastasis using preclinical models, newer and more efficacious compounds and therapies have been developed that are being evaluated (or will soon be) in clinical trails. In this chapter, we discuss novel therapeutic targets and strategies for the treatment of metastatic bone disease. Future, successful treatment of skeletal metastasis will rely on targeting critical molecular mediators/processes in both metastasis-initiating subpopulations of osteotropic cancers ("the seed") together with their supportive, cellular and extra-cellular surrounding bone/bone marrow stroma ("the soil").
Asunto(s)
Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Neoplasias Óseas/tratamiento farmacológico , Neoplasias Óseas/secundario , Diseño de Fármacos , Animales , Conservadores de la Densidad Ósea/farmacología , Conservadores de la Densidad Ósea/uso terapéutico , Médula Ósea/efectos de los fármacos , Médula Ósea/metabolismo , Neoplasias Óseas/metabolismo , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/fisiopatología , Difosfonatos/farmacología , Difosfonatos/uso terapéutico , Femenino , Humanos , Masculino , Redes y Vías Metabólicas/efectos de los fármacos , Células Madre Neoplásicas/efectos de los fármacos , Células Madre Neoplásicas/metabolismo , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/fisiopatologíaRESUMEN
Erythrocytes contain a plasma membrane redox system that can reduce extracellular ascorbate radicals by using intracellular ascorbate as an electron donor. In this study, the hypothesis was tested that cytochrome b561 was a component of this system. Spectroscopic analysis of erythrocyte membrane preparations revealed the presence of cytochrome b5 and hemoglobin but also of a cytochrome with properties similar to cytochrome b561, reducible by ascorbate and insensitive to CO. The presence of cytochrome b561 was studied further by reverse transcriptase-PCR analysis of erythrocyte progenitor cells, reticulocytes. However, no cytochrome b561 mRNA could be found. These results were corroborated by Western blot analysis with an anti-cytochrome b561 serum. No cytochrome b561 protein could be detected in extracts of erythrocyte membranes. It is therefore concluded that erythrocytes do not contain cytochrome b561 in their membranes. The possible involvement of other b-cytochromes in ascorbate-ascorbate free radical oxidoreductase activity is discussed.