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1.
Int J Mol Sci ; 24(8)2023 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-37108208

RESUMO

Glioblastoma (GBM) is the most common and malignant primary brain cancer in adults. Without treatment the mean patient survival is approximately 6 months, which can be extended to 15 months with the use of multimodal therapies. The low effectiveness of GBM therapies is mainly due to the tumor infiltration into the healthy brain tissue, which depends on GBM cells' interaction with the tumor microenvironment (TME). The interaction of GBM cells with the TME involves cellular components such as stem-like cells, glia, endothelial cells, and non-cellular components such as the extracellular matrix, enhanced hypoxia, and soluble factors such as adenosine, which promote GBM's invasiveness. However, here we highlight the role of 3D patient-derived glioblastoma organoids cultures as a new platform for study of the modeling of TME and invasiveness. In this review, the mechanisms involved in GBM-microenvironment interaction are described and discussed, proposing potential prognosis biomarkers and new therapeutic targets.


Assuntos
Neoplasias Encefálicas , Glioblastoma , Adulto , Humanos , Glioblastoma/terapia , Glioblastoma/patologia , Neoplasias Encefálicas/terapia , Neoplasias Encefálicas/patologia , Células Endoteliais/patologia , Encéfalo/patologia , Matriz Extracelular/patologia , Microambiente Tumoral , Linhagem Celular Tumoral
2.
Cells ; 12(3)2023 02 03.
Artigo em Inglês | MEDLINE | ID: mdl-36766848

RESUMO

Glioblastoma (GBM) is the most common and aggressive type of brain tumor due to its elevated recurrence following treatments. This is mainly mediated by a subpopulation of cells with stemness traits termed glioblastoma stem-like cells (GSCs), which are extremely resistant to anti-neoplastic drugs. Thus, an advancement in the understanding of the molecular processes underlying GSC occurrence should contribute significantly towards progress in reducing aggressiveness. High levels of endothelin-converting enzyme-1 (ECE1), key for endothelin-1 (ET-1) peptide activation, have been linked to the malignant progression of GBM. There are four known isoforms of ECE1 that activate ET-1, which only differ in their cytoplasmic N-terminal sequences. Isoform ECE1c is phosphorylated at Ser-18 and Ser-20 by protein kinase CK2, which increases its stability and hence promotes aggressiveness traits in colon cancer cells. In order to study whether ECE1c exerts a malignant effect in GBM, we designed an ECE1c mutant by switching a putative ubiquitination lysine proximal to the phospho-serines Lys-6-to-Arg (i.e., K6R). This ECE1cK6R mutant was stably expressed in U87MG, T98G, and U251 GBM cells, and their behavior was compared to either mock or wild-type ECE1c-expressing clone cells. ECE1cK6R behaved as a highly stable protein in all cell lines, and its expression promoted self-renewal and the enrichment of a stem-like population characterized by enhanced neurospheroid formation, as well as increased expression of stem-like surface markers. These ECE1cK6R-derived GSC-like cells also displayed enhanced resistance to the GBM-related chemotherapy drugs temozolomide and gemcitabine and increased expression of the ABCG2 efflux pump. In addition, ECE1cK6R cells displayed enhanced metastasis-associated traits, such as the modulation of adhesion and the enhancement of cell migration and invasion. In conclusion, the acquisition of a GSC-like phenotype, together with heightened chemoresistance and invasiveness traits, allows us to suggest phospho-ECE1c as a novel marker for poor prognosis as well as a potential therapeutic target for GBM.


Assuntos
Glioblastoma , Humanos , Glioblastoma/metabolismo , Enzimas Conversoras de Endotelina/genética , Enzimas Conversoras de Endotelina/metabolismo , Linhagem Celular Tumoral , Células-Tronco Neoplásicas/patologia , Fenótipo
3.
Int J Mol Sci ; 23(16)2022 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-36012307

RESUMO

Glioblastoma is the most common and aggressive primary brain tumor, characterized by its high chemoresistance and the presence of a cell subpopulation that persists under hypoxic niches, called glioblastoma stem-like cells (GSCs). The chemoresistance of GSCs is mediated in part by adenosine signaling and ABC transporters, which extrude drugs outside the cell, such as the multidrug resistance-associated proteins (MRPs) subfamily. Adenosine promotes MRP1-dependent chemoresistance under normoxia. However, adenosine/MRPs-dependent chemoresistance under hypoxia has not been studied until now. Transcript and protein levels were determined by RT-qPCR and Western blot, respectively. MRP extrusion capacity was determined by intracellular 5 (6)-Carboxyfluorescein diacetate (CFDA) accumulation. Cell viability was measured by MTS assays. Cell cycle and apoptosis were determined by flow cytometry. Here, we show for the first time that MRP3 expression is induced under hypoxia through the A2B adenosine receptor. Hypoxia enhances MRP-dependent extrusion capacity and the chemoresistance of GSCs. Meanwhile, MRP3 knockdown decreases GSC viability under hypoxia. Downregulation of the A2B receptor decreases MRP3 expression and chemosensibilizes GSCs treated with teniposide under hypoxia. These data suggest that hypoxia-dependent activation of A2B adenosine receptor promotes survival of GSCs through MRP3 induction.


Assuntos
Neoplasias Encefálicas , Glioblastoma , Proteínas Associadas à Resistência a Múltiplos Medicamentos , Adenosina/metabolismo , Neoplasias Encefálicas/metabolismo , Resistencia a Medicamentos Antineoplásicos , Glioblastoma/metabolismo , Humanos , Hipóxia/metabolismo , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , Células-Tronco Neoplásicas/metabolismo , Receptor A2B de Adenosina/metabolismo , Receptores Purinérgicos P1/metabolismo
4.
Biology (Basel) ; 11(2)2022 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-35205179

RESUMO

Glioblastoma (GBM) is the most frequent and aggressive brain tumor, characterized by great resistance to treatments, as well as inter- and intra-tumoral heterogeneity. GBM exhibits infiltration, vascularization and hypoxia-associated necrosis, characteristics that shape a unique microenvironment in which diverse cell types are integrated. A subpopulation of cells denominated GBM stem-like cells (GSCs) exhibits multipotency and self-renewal capacity. GSCs are considered the conductors of tumor progression due to their high tumorigenic capacity, enhanced proliferation, invasion and therapeutic resistance compared to non-GSCs cells. GSCs have been classified into two molecular subtypes: proneural and mesenchymal, the latter showing a more aggressive phenotype. Tumor microenvironment and therapy can induce a proneural-to-mesenchymal transition, as a mechanism of adaptation and resistance to treatments. In addition, GSCs can transition between quiescent and proliferative substates, allowing them to persist in different niches and adapt to different stages of tumor progression. Three niches have been described for GSCs: hypoxic/necrotic, invasive and perivascular, enhancing metabolic changes and cellular interactions shaping GSCs phenotype through metabolic changes and cellular interactions that favor their stemness. The phenotypic flexibility of GSCs to adapt to each niche is modulated by dynamic epigenetic modifications. Methylases, demethylases and histone deacetylase are deregulated in GSCs, allowing them to unlock transcriptional programs that are necessary for cell survival and plasticity. In this review, we described the effects of GSCs plasticity on GBM progression, discussing the role of GSCs niches on modulating their phenotype. Finally, we described epigenetic alterations in GSCs that are important for stemness, cell fate and therapeutic resistance.

5.
Front Oncol ; 10: 1665, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32974206

RESUMO

Glioblastoma (GBM) is the most devastating and least treatable brain tumor with median survival <15 months and extremely high recurrence rates. Promising results of immune checkpoint blockade obtained from pre-clinical studies in mice did not translate to clinic, and new strategies are urgently needed, particularly those targeting GBM stem cells (GSCs) that are held responsible for drug resistance and tumor recurrence. Patient-derived GSC cultures are critical for finding effective brain tumor therapies. Here, we investigated the ability of the recently described monoclonal antibody Nilo1 to specifically recognize GSCs isolated from GBM surgical samples. We employed five patient-derived GSC cultures with different stemness marker expression and differentiation potential, able to recapitulate original tumors when xenotransplanted in vivo. To answer whether Nilo1 has any functional effects in patient-derived GSCs lines, we treated the cells with Nilo1 in vitro and analyzed cell proliferation, cell cycle, apoptosis, sphere formation, as well as the expression of stem vs. differentiation markers. All tested GSCs stained positively for Nilo1, and the ability of Nilo1 to recognize GSCs strongly relied on their stem-like phenotype. Our results showed that a subset of patient-derived GSCs were sensitive to Nilo1 treatment. In three GSC lines Nilo1 triggered differentiation accompanied by the induction of p21. Most strikingly, in one GSC line Nilo1 completely abrogated self-renewal and led to Bax-associated apoptosis. Our data suggest that Nilo1 targets a molecule functionally relevant for stemness maintenance and pinpoint Nilo1 as a novel antibody-based therapeutical strategy to be used either alone or in combination with cytotoxic drugs for GSC targeting. Further pre-clinical studies are needed to validate the effectiveness of GSC-specific Nilo1 targeting in vivo.

6.
Cells ; 9(8)2020 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-32824670

RESUMO

Glioblastoma multiforme is one of the most malignant types of cancer. This is mainly due to a cell subpopulation with an extremely aggressive potential, called glioblastoma stem-like cells (GSCs). These cells produce high levels of extracellular adenosine which has been associated with increased chemoresistance, migration, and invasion in glioblastoma. In this study, we attempted to elucidate the mechanisms that control extracellular adenosine levels in GSC subtypes. By using primary and U87MG-derived GSCs, we associated increased extracellular adenosine with the mesenchymal phenotype. [3H]-adenosine uptake occurred mainly through the equilibrative nucleoside transporters (ENTs) in GSCs, but mesenchymal GSCs have lower expression and ENT1-mediated uptake activity than proneural GSCs. By analyzing expression and enzymatic activity, we determined that ecto-5'-nucleotidase (CD73) is predominantly expressed in proneural GSCs, driving AMPase activity. While in mesenchymal GSCs, both CD73 and Prostatic Acid Phosphatase (PAP) contribute to the AMP (adenosine monophosphate) hydrolysis. We did not observe significant differences between the expression of proteins involved in the metabolization of adenosine among the GCSs subtypes. In conclusion, the lower expression and activity of the ENT1 transporter in mesenchymal GSCs contributes to the high level of extracellular adenosine that these GSCs present.


Assuntos
Adenosina/metabolismo , Neoplasias Encefálicas/metabolismo , Transportador Equilibrativo 1 de Nucleosídeo/metabolismo , Espaço Extracelular/metabolismo , Glioblastoma/metabolismo , Células-Tronco Neoplásicas/metabolismo , 5'-Nucleotidase/metabolismo , Fosfatase Ácida/metabolismo , Transporte Biológico , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Proteínas Ligadas por GPI/metabolismo , Glioblastoma/patologia , Humanos
7.
Cells ; 8(11)2019 10 30.
Artigo em Inglês | MEDLINE | ID: mdl-31671624

RESUMO

Glioblastoma is the brain tumor with the worst prognosis. This is mainly due to a cell subpopulation with an extremely aggressive potential, called glioblastoma stem-like cells (GSCs). These cells produce high levels of extracellular adenosine, which are increased even more under hypoxic conditions. Under hypoxia, adenosine signaling is related to HIF-2α expression, enhancing cell aggressiveness. Adenosine can be degraded using recombinant adenosine deaminase (ADA) to revert its pathological effects. The aim of this study was to degrade adenosine using ADA in order to decrease malignancy of GSCs. Adenosine depletion was performed using recombinant ADA. Migration and invasion were measured by transwell and matrigel-coated transwell assay, respectively. HIF-2α-dependent cell migration/invasion decreased in GSCs treated with ADA under hypoxia. MRPs-mediated chemoresistance and colony formation decreased in treatment with ADA. In conclusion, adenosine depletion using adenosine deaminase decreases GSCs aggressiveness.


Assuntos
Adenosina/deficiência , Neoplasias Encefálicas/patologia , Movimento Celular , Proliferação de Células , Resistencia a Medicamentos Antineoplásicos , Glioblastoma/patologia , Células-Tronco Neoplásicas/patologia , Adenosina/metabolismo , Antineoplásicos Fitogênicos/farmacologia , Apoptose , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/metabolismo , Adesão Celular , Glioblastoma/tratamento farmacológico , Glioblastoma/metabolismo , Humanos , Hipóxia , Invasividade Neoplásica , Células-Tronco Neoplásicas/efeitos dos fármacos , Células-Tronco Neoplásicas/metabolismo , Células Tumorais Cultivadas , Vincristina/farmacologia
8.
Cancer Lett ; 446: 112-122, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30660649

RESUMO

Glioblastoma (GBM) is the brain tumor with the worst prognosis composed of a cell subpopulation called Glioblastoma Stem-like Cells (GSCs) responsible for tumor recurrence mediated by cell invasion. GSCs persist in a hypoxic microenvironment which promotes extracellular adenosine production and activation of the A3 Adenosine Receptor (A3AR), therefore, the aim of this study was to determine the role of extracellular adenosine and A3AR on GSCs invasion under hypoxia. GSCs were obtained from a U87MG cell line and primary cultures of GBM patients, and then incubated under normoxia or hypoxia. Gene expression was evaluated by RNAseq, RT-qPCR, and western blot. Cell migration was measured by spreading and transwell boyden chamber assays; cell invasion was evaluated by Matrigel-coated transwell, ex vivo brain slice, and in vivo xenograft assays. The contribution of A3AR on cell migration/invasion was evaluated using the A3AR antagonist, MRS1220. Extracellular adenosine production was higher under hypoxia than normoxia, mainly by the catalytic action of the prostatic acid phosphatase (PAP), promoting cell migration/invasion in a HIF-2-dependent process. A3AR blockade decreased cell migration/invasion and the expression of Epithelial-Mesenchymal Transition markers. In conclusion, high levels of extracellular adenosine production enhance cell migration/invasion of GSCs, through HIF-2/PAP-dependent activation of A3AR under hypoxia.


Assuntos
Adenosina/metabolismo , Neoplasias Encefálicas/metabolismo , Movimento Celular , Glioblastoma/metabolismo , Células-Tronco Neoplásicas/metabolismo , Receptor A3 de Adenosina/metabolismo , Fosfatase Ácida/genética , Fosfatase Ácida/metabolismo , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patologia , Linhagem Celular Tumoral , Glioblastoma/genética , Glioblastoma/patologia , Humanos , Camundongos Endogâmicos NOD , Camundongos SCID , Invasividade Neoplásica , Células-Tronco Neoplásicas/patologia , Receptor A3 de Adenosina/genética , Transdução de Sinais , Células Tumorais Cultivadas , Hipóxia Tumoral , Microambiente Tumoral
9.
J Glob Oncol ; 4: 1-7, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30241220

RESUMO

PURPOSE: Incidence and prevalence are important factors in policy making and planning in health care systems. The aim of this study was to compare two different estimates of the incidence and prevalence of cancer in Colombia-real-world data from the health care system and estimates from cancer registries. MATERIALS AND METHODS: Data from all providers were aggregated by the High-Cost Diseases Office (Cuenta de Alto Costo [CAC]). The real-world, age-standardized observed incidence (OI) and observed prevalence (OP) rates were calculated using the number of patients with a diagnosis of cancer who were cared for in the national health system between 2014 and 2015. The registry estimated incidence (EI) and estimated prevalence (EP) were extracted from GLOBOCAN population fact sheets for 2012, which use data from four Colombian city-based registries and extrapolate survival using the average for Asian countries, together with registries from Uganda and Zimbabwe. RESULTS: A total of 130,441 patients were analyzed. The OI of cancer in Colombia was 69.2 and the OP was 479 (per 100,000 people) in early 2015, whereas the EI was 175.2 and the 5-year EP was 501.2 (per 100,000 people), showing a higher estimate from GLOBOCAN data for 2012 than was observed in early 2015 by the CAC. Some differences were higher in specific cancers. CONCLUSION: Because of differences in methodology, the EI and the EP are not comparable to the OI and the OP. Policymakers need robust and current information to prioritize disease prevention and control programs. In Colombia, the OI and the OP-calculated by the CAC with data from the whole country-offer an opportunity for a more precise real-world estimation of patients with cancer in Colombia.


Assuntos
Neoplasias/epidemiologia , Colômbia/epidemiologia , Feminino , Humanos , Incidência , Masculino , Prevalência
10.
Int J Mol Sci ; 19(4)2018 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-29670017

RESUMO

Glioblastoma (GBM) is a neoplasm characterized by an extensive blood vessel network. Hypoxic niches of GBM can induce tumorigenic properties of a small cell subpopulation called Glioblastoma stem-like cells (GSCs) and can also increase extracellular adenosine generation which activates the A3 adenosine receptor (A3AR). Moreover, GSCs potentiates the persistent neovascularization in GBM. The aim of this study was to determine if A3AR blockade can reduce the vasculogenesis mediated by the differentiation of GSCs to Endothelial Cells (ECs) under hypoxia. We evaluated the expression of endothelial cell markers (CD31, CD34, CD144, and vWF) by fluorescence-activated cell sorting (FACS), and vascular endothelial growth factor (VEGF) secretion by ELISA using MRS1220 (A3AR antagonist) under hypoxia. We validate our results using U87MG-GSCs A3AR knockout (GSCsA3-KO). The effect of MRS1220 on blood vessel formation was evaluated in vivo using a subcutaneous GSCs-tumor model. GSCs increased extracellular adenosine production and A3AR expression under hypoxia. Hypoxia also increased the percentage of GSCs positive for endothelial cell markers and VEGF secretion, which was in turn prevented when using MRS1220 and in GSCsA3-KO. Finally, in vivo treatment with MRS1220 reduced tumor size and blood vessel formation. Blockade of A3AR decreases the differentiation of GSCs to ECs under hypoxia and in vivo blood vessel formation.


Assuntos
Diferenciação Celular , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Glioblastoma/metabolismo , Glioblastoma/patologia , Receptor A3 de Adenosina/metabolismo , Adenosina/farmacologia , Antagonistas do Receptor A3 de Adenosina/farmacologia , Animais , Biomarcadores Tumorais/metabolismo , Diferenciação Celular/efeitos dos fármacos , Hipóxia Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Células Endoteliais/efeitos dos fármacos , Humanos , Masculino , Modelos Biológicos , Células-Tronco Neoplásicas/efeitos dos fármacos , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Neovascularização Fisiológica/efeitos dos fármacos , Ratos Sprague-Dawley
11.
Mol Aspects Med ; 60: 38-51, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29222067

RESUMO

The role of extracellular vesicles in cancer biology has emerged as a focus of the study of great importance and has been shown to directly influence tumour development in several cancers including brain tumours, such as gliomas. Gliomas are the most aggressive brain tumours, and in the last time, a considerable effort has been made to understand their biology. Studies focus in the signalling pathways involved in the processes of angiogenesis, viability, drug resistance and immune response evasion, as well as gliomas ability to infiltrate healthy tissue, a phenomenon regulated by the migratory and invasive capacity of the cells within a tumour. In this review, we summarize the different types and classifications of extracellular vesicles, their intravesicular content, and their role in the regulation of tumour progression processes in glioma.


Assuntos
Neoplasias Encefálicas/diagnóstico , Neoplasias Encefálicas/metabolismo , Vesículas Extracelulares/metabolismo , Glioblastoma/diagnóstico , Glioblastoma/metabolismo , Animais , Biomarcadores Tumorais , Micropartículas Derivadas de Células/metabolismo , Progressão da Doença , Exossomos/metabolismo , Humanos , Microambiente Tumoral
12.
Curr Med Chem ; 24(25): 2781-2795, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28260500

RESUMO

BACKGROUND: In the last two decades, there have been significant technological advances in the early detection of brain tumors. However, no notable improvements have been observed in the treatment of Glioblastoma Multiforme (GBM), the most common brain neoplasm coupled with the worst prognosis. GBM is characterized by an extensive resistance to a broad spectrum of anti-tumor drugs. This property is the result of a phenomenon known as Multiple Drug Resistance (MDR), which significantly limits noninvasive alternative therapies. This limitation is primarily due to the activity of ABC transporters and proteins related with DNA repair such as the MGMT enzyme. Due to the high mortality rate in GBM patients and current treatment deficits, new therapeutic strategies for this type of neoplasm are of vital importance. METHODS: In this review, proposed treatments for GBM, including the use of alkylating agents with MGMT inhibitors, MDR modulators, and immunotherapies are discussed. We focused our bibliographic research on papers containing in vitro, in vivo, and clinical phase analysis published over the last 20 years. RESULTS: Several studies have demonstrated good results using alkylating agents plus MGMT inhibitors, although without great improvements in survival. The use of modulators of ABC transporters enhances the effects of chemotherapy, proving it an effective complementary therapy. Immunotherapies have undergone significant developments as a directed and personalized approach for GBM treatment. CONCLUSION: The use of alternative complementary therapies discussed in this review could increase the survival of GBM patients; however, additional clinical phase analysis and the generation of new treatment protocols are required.


Assuntos
Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Glioblastoma/tratamento farmacológico , Antineoplásicos/química , Humanos
13.
Mol Aspects Med ; 55: 140-151, 2017 06.
Artigo em Inglês | MEDLINE | ID: mdl-28223127

RESUMO

Glioblastoma multiforme (GBM) is considered the most common and aggressive tumour of the central nervous system and is characterized for being highly chemoresistant. This property is mainly due to the activation of Multiple Drug Resistance (MDR) mechanisms that protect cancer cells from structurally and morphologically different drugs. Overexpression and increased ABC transporters activity is one of the most important MDR mechanisms at the clinical level, and both its expression and activity are elevated in GBM cells. Within the tumour, there is a subpopulation called glioblastoma stem-like cells (GSCs), which due to its high tumourigenic capacity and chemoresistance, have been postulated as the main responsible for tumour recurrence. The GSCs inhabit hypoxic tumour zones, niches that apart from maintaining and promoting stem phenotype have also been correlated with high chemoresistance. Of the signalling pathways activated during hypoxia, purinergic signalling has been highly associated to the induction of MDR mechanisms. Through its receptors, the nucleoside adenosine has been shown to promotes the chemoresistance mediated by ABC transporters. Therefore, targeting its components is a promising alternative for GBM treatment. In this review, we will discuss chemoresistance in GSCs and the effect of the hypoxic microenvironment and adenosine on MDR mechanisms.


Assuntos
Adenosina/genética , Resistência a Múltiplos Medicamentos/genética , Resistencia a Medicamentos Antineoplásicos/genética , Glioblastoma/tratamento farmacológico , Transportadores de Cassetes de Ligação de ATP/genética , Adenosina/metabolismo , Hipóxia Celular/genética , Glioblastoma/genética , Glioblastoma/patologia , Humanos , Células-Tronco Neoplásicas/efeitos dos fármacos , Transdução de Sinais , Microambiente Tumoral/genética
14.
Oncotarget ; 7(41): 67373-67386, 2016 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-27634913

RESUMO

MRP1 transporter correlates positively with glioma malignancy and the Multiple Drug Resistance (MDR) phenotype in Glioblastoma Multiforme (GBM). Evidence shows that the MRP1 transporter is controlled by the adenosine signalling axis. The aim of this study was to identify the role of adenosine on the MDR phenotype in Glioblastoma Stem-like Cells (GSCs), the cell population responsible for the tumorigenic and chemoresistance capabilities of this tumour. We found that GSCs have increased intrinsic capacity to generate extracellular adenosine, thus controlling MRP1 transporter expression and activity via activation of the adenosine A3 receptor (A3AR). We showed PI3K/Akt and MEK/ERK1/2 signaling pathways downstream A3AR to control MRP1 in GSCs. In vitro pharmacological blockade of A3AR had a chemosensitizing effect, enhancing the actions of antitumour drugs and decreasing cell viability and proliferation of GSCs. In addition, we produced an in vivo xenograft model by subcutaneous inoculation of human GSCs in NOD/SCID-IL2Rg null mice. Pharmacological blockade of A3AR generated a chemosensitizing effect, enhancing the effectiveness of the MRP1 transporter substrate, vincristine, reducing tumour size and the levels of CD44 and Nestin stem cell markers as well as the Ki-67 proliferation indicator. In conclusion, we demonstrated the chemosensitizing effect of A3AR blockade on GSCs.


Assuntos
Neoplasias Encefálicas/patologia , Resistencia a Medicamentos Antineoplásicos/fisiologia , Glioblastoma/patologia , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , Células-Tronco Neoplásicas/patologia , Receptor A3 de Adenosina/metabolismo , Animais , Neoplasias Encefálicas/metabolismo , Glioblastoma/metabolismo , Xenoenxertos , Humanos , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Células-Tronco Neoplásicas/metabolismo
15.
J Neurooncol ; 128(1): 9-19, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-26900077

RESUMO

The most aggressive type of brain tumor is glioblastoma multiforme, which to date remains incurable. Thuja occidentalis is used in homeopathy for the treatment of cancer, however, its mechanism of action remains unknown. We set out to study the effects of thujone fractions of Thuja on glioblastoma using in vitro and in vivo models. We found that the α/ ß-thujone fraction decrease the cell viability and exhibit a potent anti-proliferative, pro-apoptotic and anti-angiogenic effects in vitro. In vivo assays showed that α /ß-thujone promotes the regression of neoplasia and inhibits the angiogenic markers VEGF, Ang-4 and CD31 into the tumor.


Assuntos
Antineoplásicos/farmacologia , Glioblastoma/tratamento farmacológico , Monoterpenos/farmacologia , Fitoterapia , Extratos Vegetais/farmacologia , Thuja , Inibidores da Angiogênese/farmacologia , Animais , Apoptose/efeitos dos fármacos , Monoterpenos Bicíclicos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Glioblastoma/irrigação sanguínea , Glioblastoma/patologia , Glioblastoma/fisiopatologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/fisiologia , Humanos , Masculino , Transplante de Neoplasias , Ratos Sprague-Dawley
16.
Arch. med ; 13(2): 220-225, 30/dez. 2013.
Artigo em Espanhol | LILACS | ID: lil-707524

RESUMO

Objetivo: describir las características clínicas de los pacientes con anemia hemolítica autoinmune en una clínica privada de la ciudad de Medellín. Materiales y métodos: Estudio descriptivo retrospectivo de siete casos AHAl que fueron atendidos en una clínica privada de la ciudad de Medellín (Colombia) entre el 2005 y 2010. La información fue tomada de la historia clínica de los pacientes. Resultados: 6 de los 7 casos eran mujeres, la edad mediana fue 47 años (mínima 21 y máxima 74). La enfermedades concomitantes más frecuentes fueron LES y las infecciosas con 3 casos cada una. La terapia se basó principalmente en el uso de glucocorticoides con 4 casos hallados y nunca se recurrió a la terapia biológica. Dos pacientes requirieron cambio de tratamiento.Conclusión: Este estudio muestra los hallazgos clínicos y demográficos de una pequeña serie de casos adultos con AHAI. Esta frecuencia tan baja probablemente está condicionada a la baja incidencia de esta enfermedad en el medio. Es necesario realizar estudios nacionales de cobertura geográfica mayor que ayuden a establecer la real incidencia de esta patología y describir las principales características clínicas y epidemiológicas.


Assuntos
Anemia Hemolítica Autoimune , Anticorpos , Autoimunidade
17.
Acta biol. colomb ; 14(3): 181-198, dic. 2009.
Artigo em Espanhol | LILACS | ID: lil-634925

RESUMO

La filósfera es un ecosistema complejo que a pesar de contar con un gran número de microorganismos asociados, presenta pocos estudios diseñados para entender las características ecofisiológicas de los microorganismos que lo habitan. Las levaduras, como elemento importante de este hábitat, están expuestas a un ambiente variable afectado por la planta hospedera, edad y posición de la hoja, disponibilidad y calidad de nutrientes, temperatura, pH, radiación y actividad del agua. Estos factores producen una presión de selección para el establecimiento de poblaciones naturales o introducidas de levaduras que puedan desplazar otras poblaciones de hongos fitopatógenos. En este estudio se obtuvieron 80 aislamientos de levaduras filosféricas de dos cultivos de mora (Rubus glaucus), a las que se les determinó su nicho potencial en términos de su capacidad para crecer a diferentes condiciones de temperatura, pH, estrés osmótico y tolerancia a radiación UV. El estrés osmótico fue la condición evaluada más restrictiva para las levaduras obtenidas, ya que únicamente seis aislamientos presentaron un crecimiento superior a 0,3 unidades de DO a 405 nm, cuando fueron crecidas a 50% y 60% de glucosa. Este estudio permitió seleccionar diez aislamientos filosféricos de levaduras, destacados por su capacidad para crecer en un amplio rango de condiciones. De estos, cuatro aislamientos LvF 34, LvF 43, LvF 44 y LvF 50 se destacaron por su capacidad de antagonismo contra el hongo fitopatógeno Botritys cinérea. Su determinación taxonómica permitió reportar por primera vez las especies Candida kunwinensis y Rhodotorula colostri con potencial biocontrolador.


The phyllosphere is a complex ecosystem in which despite of having a great number of associated microorganisms, few studies have been designed to understand the ecophysiological characteristics of the microorganism’s inhabitant. Yeasts, as an important element of this habitat, are exposed to a variable environment, affected by the host plant, age and leaf position, availability and nutrient quality, temperature, pH, radiation and water activity. All these factors produce an important selection pressure for the establishment of natural or introduced populations of yeast which may displace other phytopathogenic fungi. In this study 80 isolates of phyllospheric yeast were obtained from two blackberry crops (Rubus glaucus). Its potential niche was determined in terms of its capacity to growth at different conditions of temperature, pH, osmotic stress and UV radiation. The osmotic stress was the most restrictive evaluated condition for the obtained yeast. Only six isolates presented a growth higher to 0.3 OD units at 405 nm, when grown at 50% or 60% of glucose. This study identified the ecological niche of ten yeast phyllospheric isolates, selected for their capacity to growth at a wide range of conditions. Four of these isolates LvF 34, LvF 43, LvF 44 and LvF 50 were selected for their antagonistic capacity against the phytopathogenic fungi Botritys cinérea. Its taxonomic determination allowed us to report for the first time isolates of Candida kunwinensis and Rhodotorula colostri with biocontrol capacity.

18.
Acta biol. colomb ; 12(1): 19-32, jun. 2007. tab, graf
Artigo em Inglês | LILACS | ID: lil-468988

RESUMO

Rhizoctonia solani is a soil borne phytopathogen associated with reduced plant vigor and tuber production in potato crops. There is a huge interest to search alternatives of biological control management of this disease, because the potato crops in Colombia are the highest consumers of chemical pesticides in Colombia. In order to obtain a fluorescent Pseudomonas strain with the capacity to reduce the disease symptoms produced by R. solani, determination and isolation of the predominant fluorescent Pseudomonas in several potato crops of the main Colombian producing region was done in a previous study. Six different P. fluorescens strains with none, moderate and high fungal growth inhibition capacity in vitro, were used in this study. Despite of the differences found in the dynamics of colonization and colonization capacity, all evaluated strains induced S. phureja growth and reduced disease symptoms produced by R. solani. Our results support the conclusion that association of P. fluorescens strains with S. phureja rhizosphere is a feasible alternative for the management of R. solani symptoms.Rhizoctonia solani es un hongo fitopatógeno del suelo, el cual produce una reducción significativa del vigor de las plantas y de la producción de tubérculos en cultivos de papa. Es de gran interés la búsqueda de alternativas de manejo de esta enfermedad, especialmente desde la perspectiva de control biológico ya que los cultivos de papa son los mayores consumidores de plaguicidas de origen químicos en Colombia. Con el objeto de obtener una cepa del grupo de las Pseudomonas fluorescentes con la capacidad para reducir los síntomas de la enfermedad producidos por R. solani, se realizó en un estudio previo el aislamiento y caracterización de una colección de aislamientos de Pseudomonas fluorescentes provenientes de diferentes cultivos de la región papera más productiva del país. Seis cepas nativas de P. fluorescens con buena, moderada o ninguna capacidad para inhibir...


Assuntos
Fungos/patogenicidade , Praguicidas/intoxicação , Tubérculos/parasitologia
19.
Rev. colomb. biotecnol ; 2(1): 50-58, jul. 1999. tab, graf
Artigo em Espanhol | LILACS | ID: lil-506977

RESUMO

Las pseudomonas fluorescentes son unas de las bacterias benéficas más importantes a nivel de la rizosfera gracias a que pueden controlar algunos fitopatógenos habituales del suelo como resultado de su capacidad antagonista. Hay muy pocos trabajos realizados para conocer la composi-ción y diversidad de pseudomonas fluorescentes en paí-ses tropicales. En este trabajo se determinó la composi-ción de pseudomonas fluorescentes provenientes de dife-rentes cultivos de papa ubicados en la región Cundiboyacense, entre 2.100 y 3.200 msnm, la cual es considerada la zona de mayor producción de papa de Colombia. Así mismo se evaluó el efecto de algunas prác-ticas de cultivo sobre dicha composición. Finalmente se determinó la capacidad de antagonismo de algunos aisla-mientos de pseudomonas fluorescentes contra Rhizoctonia solani. Fueron evaluadas 45 muestras de rizosfera y rizoplano de cultivos de papa provenientes de 15 campos de papa diferentes. Las muestras se procesaron usando una modificación del medio King B, la cual resultó ser más eficiente para aislar pseudomonas fluorescentes, que otras reportadas en la literatura. El 80,7por cien y 82,7por cien de las pseudomonas fluorescentes aisladas en rizosfera y rizoplano, respectivamente, fueron Pseudomonas fluorescens, mostrando poca diversidad en las muestras evaluadas. Por otra parte se observó un efecto de la estra-tegia de rotación de cultivo maíz-papa, sobre el número de pseudomonas fluorescentes aisladas de la rizosfera. Estos resultados tienen implicaciones para el diseño de estrategias de manejo integrado de plagas en los cultivos de los países tropicales.


Assuntos
Microbiologia de Alimentos , Microbiologia , Pseudomonas fluorescens , Rhizoctonia , Solanum tuberosum
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