Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 8.527
Filtrar
1.
Int J Mol Sci ; 22(6)2021 Mar 17.
Artículo en Inglés | MEDLINE | ID: mdl-33802972

RESUMEN

Leukaemia is a malignant disease of the blood. Current treatments for leukaemia are associated with serious side-effects. Plant-derived polyphenols have been identified as potent anti-cancer agents and have been shown to work synergistically with standard chemotherapy agents in leukaemia cell lines. Polyphenols have multiple mechanisms of action and have been reported to decrease cell proliferation, arrest cell cycle and induce apoptosis via the activation of caspase (3, 8 and 9); the loss of mitochondrial membrane potential and the release of cytochrome c. Polyphenols have been shown to suppress activation of transcription factors, including NF-kB and STAT3. Furthermore, polyphenols have pro-oxidant properties, with increasing evidence that polyphenols inhibit the antioxidant activity of glutathione, causing oxidative DNA damage. Polyphenols also induce autophagy-driven cancer cell death and regulate multidrug resistance proteins, and thus may be able to reverse resistance to chemotherapy agents. This review examines the molecular mechanism of action of polyphenols and discusses their potential therapeutic targets. Here, we discuss the pharmacological properties of polyphenols, including their anti-inflammatory, antioxidant, anti-proliferative, and anti-tumour activities, and suggest that polyphenols are potent natural agents that can be useful therapeutically; and discuss why data on bioavailability, toxicity and metabolism are essential to evaluate their clinical use.


Asunto(s)
Leucemia/tratamiento farmacológico , Polifenoles/uso terapéutico , Animales , Autofagia/efectos de los fármacos , Puntos de Control del Ciclo Celular/efectos de los fármacos , Humanos , Leucemia/patología , Modelos Biológicos , Polifenoles/química , Polifenoles/farmacología , Especies Reactivas de Oxígeno/metabolismo
2.
Int J Nanomedicine ; 16: 2849-2877, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33883895

RESUMEN

Background: Exosomes are endosome-derived nano-sized vesicles that have emerged as important mediators of intercellular communication and play significant roles in various diseases. However, their applications are rigorously restricted by the limited secretion competence of cells. Therefore, strategies to enhance the production and functions of exosomes are warranted. Studies have shown that nanomaterials can significantly enhance the effects of cells and exosomes in intercellular communication; however, how palladium nanoparticles (PdNPs) enhance exosome release in human leukemia monocytic cells (THP-1) remains unclear. Therefore, this study aimed to address the effect of PdNPs on exosome biogenesis and release in THP-1 cells. Methods: Exosomes were isolated by ultracentrifugation and ExoQuickTM and characterized by dynamic light scattering, nanoparticle tracking analysis system, scanning electron microscopy, transmission electron microscopy, EXOCETTM assay, and fluorescence polarization. The expression levels of exosome markers were analyzed via quantitative reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. Results: PdNP treatment enhanced the biogenesis and release of exosomes by inducing oxidative stress, endoplasmic reticulum stress, apoptosis, and immunomodulation. The exosomes were spherical in shape and had an average diameter of 50-80 nm. Exosome production was confirmed via total protein concentration, exosome counts, acetylcholinesterase activity, and neutral sphingomyelinase activity. The expression levels of TSG101, CD9, CD63, and CD81 were significantly higher in PdNP-treated cells than in control cells. Further, cytokine and chemokine levels were significantly higher in exosomes isolated from PdNP-treated THP-1 cells than in those isolated from control cells. THP-1 cells pre-treated with N-acetylcysteine or GW4869 showed significant decreases in PdNP-induced exosome biogenesis and release. Conclusion: To our knowledge, this is the first study showing that PdNPs stimulate exosome biogenesis and release and simultaneously increase the levels of cytokines and chemokines by modulating various physiological processes. Our findings suggest a reasonable approach to improve the production of exosomes for various therapeutic applications.


Asunto(s)
Apoptosis/efectos de los fármacos , Estrés del Retículo Endoplásmico , Exosomas/metabolismo , Inmunomodulación/efectos de los fármacos , Leucemia/patología , Nanopartículas del Metal/toxicidad , Estrés Oxidativo , Paladio/toxicidad , Acetilcolinesterasa/metabolismo , Acetilcisteína/farmacología , Compuestos de Anilina/farmacología , Antioxidantes/metabolismo , Compuestos de Bencilideno/farmacología , Biomarcadores de Tumor/metabolismo , Caspasas/metabolismo , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Citocinas/metabolismo , Daño del ADN , Estrés del Retículo Endoplásmico/efectos de los fármacos , Activación Enzimática/efectos de los fármacos , Regulación Leucémica de la Expresión Génica/efectos de los fármacos , Humanos , Leucemia/sangre , Nanopartículas del Metal/ultraestructura , Mitocondrias/efectos de los fármacos , Mitocondrias/patología , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Estrés Oxidativo/efectos de los fármacos , Tamaño de la Partícula , Esfingomielina Fosfodiesterasa/metabolismo , Células THP-1
3.
Int J Mol Sci ; 22(4)2021 Feb 13.
Artículo en Inglés | MEDLINE | ID: mdl-33668652

RESUMEN

Leukemia is a hematological malignancy that originates from hematopoietic stem cells in the bone marrow. Significant progress has made in understanding its pathogensis and in establishing chemotherapy and hematopoietic stem cell transplantation therapy (HSCT). However, while the successive development of new therapies, such as molecular-targeted therapy and immunotherapy, have resulted in remarkable advances, the fact remains that some patients still cannot be saved, and resistance to treatment and relapse are still problems that need to be solved in leukemia patients. The bone marrow (BM) niche is a microenvironment that includes hematopoietic stem cells and their supporting cells. Leukemia cells interact with bone marrow niches and modulate them, not only inducing molecular and functional changes but also switching to niches favored by leukemia cells. The latter are closely associated with leukemia progression, suppression of normal hematopoiesis, and chemotherapy resistance, which is precisely the area of ongoing study. Exosomes play an important role in cell-to-cell communication, not only with cells in close proximity but also with those more distant due to the nature of exosomal circulation via body fluids. In leukemia, exosomes play important roles in leukemogenesis, disease progression, and organ invasion, and their usefulness in the diagnosis and treatment of leukemia has recently been reported. The interaction between leukemia cell-derived exosomes and the BM microenvironment has received particular attention. Their interaction is believed to play a very important role; in addition to their diagnostic value, exosomes could serve as a marker for monitoring treatment efficacy and as an aid in overcoming drug resistance, among the many problems in leukemia patients that have yet to be overcome. In this paper, we will review bone marrow niches in leukemia, findings on leukemia-derived exosomes, and exosome-induced changes in bone marrow niches.


Asunto(s)
Médula Ósea/metabolismo , Comunicación Celular , Exosomas/metabolismo , Leucemia/metabolismo , Microambiente Tumoral , Médula Ósea/patología , Exosomas/patología , Humanos , Leucemia/patología
4.
Anticancer Res ; 41(3): 1429-1438, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33788734

RESUMEN

BACKGROUND/AIM: Clinical significance of antitumour drugs is limited by multidrug resistance (MDR). We examined the effect of bioreductive activation of the anthracyclines, doxorubicin (DOX) and pirarubicin (PIRA), by cytochrome P450 reductase (CPR) on triggering apoptosis of leukaemia HL60 cells and their MDR counterparts. MATERIALS AND METHODS: Cell cycle and FAS expression were investigated by flow cytometry. DNA fragmentation was examined by electrophoretic analysis and caspase-3/8 activities were determined colorimetrically. RESULTS: Non-activated and CPR-activated forms of DOX and PIRA (IC90) had similar efficacy in provoking G2/M arrest of sensitive HL60 as well as resistant HL60/VINC and HL60/DOX cells and in causing DNA degradation. Interestingly, HL60/VINC cells were more prone to apoptosis induced by all studied forms of these drugs. However, no change in Fas expression was observed. CONCLUSION: Bioreductive activation of DOX and PIRA does not affect their ability to induce apoptosis of sensitive and resistant HL60 leukaemia cells.


Asunto(s)
Apoptosis/efectos de los fármacos , Doxorrubicina/análogos & derivados , Doxorrubicina/farmacología , Resistencia a Múltiples Medicamentos/efectos de los fármacos , Resistencia a Antineoplásicos/efectos de los fármacos , Leucemia/patología , Antineoplásicos/farmacología , Caspasa 3/metabolismo , Caspasa 8/metabolismo , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Fragmentación del ADN/efectos de los fármacos , Puntos de Control de la Fase G2 del Ciclo Celular/efectos de los fármacos , Células HL-60 , Humanos , Leucemia/metabolismo , NADPH-Ferrihemoproteína Reductasa/metabolismo
5.
Transplant Cell Ther ; 27(5): 438.e1-438.e6, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-33728417

RESUMEN

An evidence-based triage plan for cellular therapy distribution is critical in the face of emerging constraints on healthcare resources. We evaluated the impact of treatment delays related to COVID-19 on patients scheduled to undergo hematopoietic cell transplantation (HCT) or chimeric antigen receptor T-cell (CAR-T) therapy at our center. Data were collected in real time between March 19 and May 11, 2020, for patients who were delayed to cellular therapy. We evaluated the proportion of delayed patients who ultimately received cellular therapy, reasons for not proceeding to cellular therapy, and changes in disease and health status during delay. A total of 85 patients were delayed, including 42 patients planned for autologous HCT, 36 patients planned for allogeneic HCT, and 7 patients planned for CAR-T therapy. Fifty-six of these patients (66%) since received planned therapy. Five patients died during the delay. The most common reason for not proceeding to autologous HCT was good disease control in patients with plasma cell dyscrasias (75%). The most common reason for not proceeding to allogeneic HCT was progression of disease (42%). All patients with acute leukemia who progressed had measurable residual disease (MRD) at the time of delay, whereas no patient without MRD at the time of delay progressed. Six patients (86%) ultimately received CAR-T therapy, including 3 patients who progressed during the delay. For patients with high-risk disease such as acute leukemia, and particularly those with MRD at the time of planned HCT, treatment delay can result in devastating outcomes and should be avoided if at all possible.


Asunto(s)
Trasplante de Células Madre Hematopoyéticas , Inmunoterapia Adoptiva , Pandemias , Tiempo de Tratamiento , Adulto , Anciano , Aloinjertos , Amiloidosis/terapia , Anemia Aplásica/terapia , /epidemiología , Defensa Civil , Infección Hospitalaria/epidemiología , Infección Hospitalaria/prevención & control , Progresión de la Enfermedad , Práctica Clínica Basada en la Evidencia/organización & administración , Femenino , Trasplante de Células Madre Hematopoyéticas/estadística & datos numéricos , Humanos , Control de Infecciones/métodos , Transmisión de Enfermedad Infecciosa de Profesional a Paciente , Leucemia/mortalidad , Leucemia/patología , Leucemia/terapia , Masculino , Persona de Mediana Edad , Enfermedades Mielodisplásicas-Mieloproliferativas/mortalidad , Enfermedades Mielodisplásicas-Mieloproliferativas/terapia , Neoplasia Residual , Neoplasias/mortalidad , Neoplasias/terapia , Ciudad de Nueva York/epidemiología , Asignación de Recursos , Tiempo de Tratamiento/estadística & datos numéricos , Trasplante Autólogo , Triaje/organización & administración , Adulto Joven
6.
Hum Genet ; 140(6): 849-861, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33385171

RESUMEN

Mitochondrial (MT) dysfunction is a hallmark of aging and has been associated with most aging-related diseases as well as immunological processes. However, little is known about aging, lifestyle and genetic factors influencing mitochondrial DNA (mtDNA) abundance. In this study, mtDNA abundance was estimated from the weighted intensities of probes mapping to the MT genome in 295,150 participants from the UK Biobank. We found that the abundance of mtDNA was significantly elevated in women compared to men, was negatively correlated with advanced age, higher smoking exposure, greater body-mass index, higher frailty index as well as elevated red and white blood cell count and lower mortality. In addition, several biochemistry markers in blood-related to cholesterol metabolism, ion homeostasis and kidney function were found to be significantly associated with mtDNA abundance. By performing a genome-wide association study, we identified 50 independent regions genome-wide significantly associated with mtDNA abundance which harbour multiple genes involved in the immune system, cancer as well as mitochondrial function. Using mixed effects models, we estimated the SNP-heritability of mtDNA abundance to be around 8%. To investigate the consequence of altered mtDNA abundance, we performed a phenome-wide association study and found that mtDNA abundance is involved in risk for leukaemia, hematologic diseases as well as hypertension. Thus, estimating mtDNA abundance from genotyping arrays has the potential to provide novel insights into age- and disease-relevant processes, particularly those related to immunity and established mitochondrial functions.


Asunto(s)
Envejecimiento/genética , ADN Mitocondrial/genética , Genoma Mitocondrial , Enfermedades Hematológicas/genética , Hipertensión/genética , Leucemia/genética , Anciano , Envejecimiento/inmunología , Bancos de Muestras Biológicas , Índice de Masa Corporal , ADN Mitocondrial/inmunología , Recuento de Eritrocitos , Femenino , Estudio de Asociación del Genoma Completo , Enfermedades Hematológicas/epidemiología , Enfermedades Hematológicas/inmunología , Enfermedades Hematológicas/patología , Humanos , Hipertensión/epidemiología , Hipertensión/inmunología , Hipertensión/patología , Inmunidad Innata , Patrón de Herencia/inmunología , Leucemia/epidemiología , Leucemia/inmunología , Leucemia/patología , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Mitocondrias/genética , Mitocondrias/inmunología , Polimorfismo de Nucleótido Simple , Fumar/genética , Fumar/fisiopatología , Reino Unido/epidemiología
7.
Nat Struct Mol Biol ; 28(2): 190-201, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33479542

RESUMEN

NUP98 fusion proteins cause leukemia via unknown molecular mechanisms. All NUP98 fusion proteins share an intrinsically disordered region (IDR) in the NUP98 N terminus, featuring repeats of phenylalanine-glycine (FG), and C-terminal fusion partners often function in gene control. We investigated whether mechanisms of oncogenic transformation by NUP98 fusion proteins are hardwired in their protein interactomes. Affinity purification coupled to mass spectrometry (MS) and confocal imaging of five NUP98 fusion proteins expressed in human leukemia cells revealed that shared interactors were enriched for proteins involved in biomolecular condensation and that they colocalized with NUP98 fusion proteins in nuclear puncta. We developed biotinylated isoxazole-mediated condensome MS (biCon-MS) to show that NUP98 fusion proteins alter the global composition of biomolecular condensates. An artificial FG-repeat-containing fusion protein phenocopied the nuclear localization patterns of NUP98 fusion proteins and their capability to drive oncogenic gene expression programs. Thus, we propose that IDR-containing fusion proteins combine biomolecular condensation with transcriptional control to induce cancer.


Asunto(s)
Núcleo Celular/metabolismo , Proteínas de Homeodominio , Leucemia , Proteínas de Complejo Poro Nuclear , Proteínas de Fusión Oncogénica , Animales , Expresión Génica , Regulación Leucémica de la Expresión Génica , Células HEK293 , Células HL-60 , Proteínas de Homeodominio/química , Proteínas de Homeodominio/fisiología , Humanos , Leucemia/metabolismo , Leucemia/patología , Ratones , Células 3T3 NIH , Proteínas de Complejo Poro Nuclear/química , Proteínas de Complejo Poro Nuclear/fisiología , Proteínas de Fusión Oncogénica/química , Proteínas de Fusión Oncogénica/fisiología
8.
Nat Commun ; 12(1): 223, 2021 01 11.
Artículo en Inglés | MEDLINE | ID: mdl-33431820

RESUMEN

Enhancers are DNA sequences that enable complex temporal and tissue-specific regulation of genes in higher eukaryotes. Although it is not entirely clear how enhancer-promoter interactions can increase gene expression, this proximity has been observed in multiple systems at multiple loci and is thought to be essential for the maintenance of gene expression. Bromodomain and Extra-Terminal domain (BET) and Mediator proteins have been shown capable of forming phase condensates and are thought to be essential for super-enhancer function. Here, we show that targeting of cells with inhibitors of BET proteins or pharmacological degradation of BET protein Bromodomain-containing protein 4 (BRD4) has a strong impact on transcription but very little impact on enhancer-promoter interactions. Dissolving phase condensates reduces BRD4 and Mediator binding at enhancers and can also strongly affect gene transcription, without disrupting enhancer-promoter interactions. These results suggest that activation of transcription and maintenance of enhancer-promoter interactions are separable events. Our findings further indicate that enhancer-promoter interactions are not dependent on high levels of BRD4 and Mediator, and are likely maintained by a complex set of factors including additional activator complexes and, at some sites, CTCF and cohesin.


Asunto(s)
Elementos de Facilitación Genéticos , Regiones Promotoras Genéticas , Transcripción Genética , Factor de Unión a CCCTC/metabolismo , Carcinogénesis/efectos de los fármacos , Carcinogénesis/genética , Proteínas de Ciclo Celular/metabolismo , Línea Celular Tumoral , Cromatina/metabolismo , Proteínas Cromosómicas no Histona/metabolismo , Proteínas de Unión al ADN/metabolismo , Glicoles/farmacología , Histonas/metabolismo , Humanos , Leucemia/genética , Leucemia/patología , Modelos Genéticos , Unión Proteica/efectos de los fármacos , Proteínas Proto-Oncogénicas c-myc/genética , Transcripción Genética/efectos de los fármacos
9.
Phytomedicine ; 80: 153383, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33091855

RESUMEN

BACKGROUND: Caffeic acid 3,4-dihydroxyphenethyl ester (CADPE) is a natural polyphenolic ester isolated as a minor component from a water extract of the Chinese medicine Zhongjiefeng [Sarcandra glabra (Thunb.) Nakai (Chloranthaceae)] and has previously shown to have activity against solid tumors through the modulation of multiple targets or signal pathways. However, the activity and potential mechanism of CADPE against leukemia cells have not yet been characterized. PURPOSE: To investigate whether and how CADPE kills leukemia cells. METHOD: (1) The activity of CADPE inhibiting the growth of different leukemia cell lines was evaluated by MTT assay; (2) Cell cycle arrest and apoptosis induced by CADPE were determined by flow cytometry with FlowJo software for quantification; (3) The protein levels were analyzed by Western blot and ubiquitin-binding c-Myc was acquired by co-immunoprecipitation. RESULTS: CADPE exerted potent activity against different leukemia cell lines with low toxicity in normal cells. In terms of mechanism of action, CADPE promoted ubiquitin-proteasome-dependent degradation of c-Myc through activating glycogen synthase kinase-3ß (GSK3ß) and downregulating deubiquitinating enzyme USP28 to trigger the interaction of c-Myc with ubiquitin ligase Fbw7, resulting in the downregulation of cell cycle regulators and anti-apoptotic proteins and consequently, cell cycle arrest and cell apoptosis. CONCLUSION: CADPE is a novel c-Myc inhibitor with high activity and a unique mechanism for killing leukemia cells.


Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Ácidos Cafeicos/farmacología , Leucemia/tratamiento farmacológico , Apoptosis/efectos de los fármacos , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proteínas F-Box/metabolismo , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Humanos , Leucemia/metabolismo , Leucemia/patología , Proteínas Proto-Oncogénicas c-myc/metabolismo , Transducción de Señal/efectos de los fármacos , Ubiquitina/metabolismo , Ubiquitina Tiolesterasa/metabolismo
10.
Eur J Med Chem ; 211: 113095, 2021 Feb 05.
Artículo en Inglés | MEDLINE | ID: mdl-33360560

RESUMEN

Targeting epigenetic dysregulation has emerged as a valuable therapeutic strategy in cancer treatment. Especially epigenetic combination therapy of histone deacetylase inhibitors (HDACi) with established anti-cancer drugs has provided promising results in preclinical and clinical studies. The structural optimization of alkoxyamide-based class I/IIb inhibitors afforded improved analogs with potent efficacy in cisplatin-resistant head and neck carcinoma cells and bortezomib-resistant leukemia cells. The most promising HDACi showed a superior synergistic cytotoxic activity as compared to vorinostat and class I HDACi in combination with cisplatin, leading to a full reversal of the chemoresistant phenotype in head and neck cancer cell lines, as well in combination with the proteasome inhibitors (bortezomib and carfilzomib) in a panel of leukemic cell lines. Furthermore, the most valuable alkoxyamide-based HDACi exhibited strong ex vivo anticancer efficacy against primary patient samples obtained from different therapy-resistant leukemic entities.


Asunto(s)
Antineoplásicos/uso terapéutico , Apoptosis/efectos de los fármacos , Epigenómica/métodos , Neoplasias de Cabeza y Cuello/tratamiento farmacológico , Inhibidores de Histona Desacetilasas/uso terapéutico , Leucemia/tratamiento farmacológico , Antineoplásicos/farmacología , Sinergismo Farmacológico , Neoplasias de Cabeza y Cuello/patología , Inhibidores de Histona Desacetilasas/farmacología , Humanos , Leucemia/patología
11.
Pediatr Blood Cancer ; 68(1): e28714, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-32979296

RESUMEN

Aerodigestive adverse effects (AD-AE) during intravenous pentamidine (IV-P) infusion for Pneumocystis jiroveci pneumonia prophylaxis are uncommon in retrospective chart review studies. We conducted a survey in patients on IV-P, which included 31 specific questions. Twenty-five patients were included in the analysis; AD-AE were observed in 22 (88%) with recurrence of symptoms in 88% participants with subsequent infusions. Five leading symptoms were congestion (48%), lip tingling (32%), nausea (28%), tongue tingling (24%), vomiting, and throat swelling (17%); multiple symptoms were reported in 72% of the patients. In conclusion, AD-AE of IV-P infusion are common, self-limited, and tend to be recurrent.


Asunto(s)
Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/epidemiología , Leucemia/terapia , Pentamidina/efectos adversos , Pneumocystis carinii/efectos de los fármacos , Neumonía por Pneumocystis/tratamiento farmacológico , Tracto Gastrointestinal Superior/patología , Adolescente , Adulto , Antifúngicos/administración & dosificación , Antifúngicos/efectos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Niño , Preescolar , Terapia Combinada , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/etiología , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/patología , Femenino , Estudios de Seguimiento , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Humanos , Infusiones Intravenosas , Leucemia/patología , Masculino , Michigan/epidemiología , Pentamidina/administración & dosificación , Neumonía por Pneumocystis/etiología , Neumonía por Pneumocystis/patología , Pronóstico , Autoinforme , Encuestas y Cuestionarios , Tracto Gastrointestinal Superior/efectos de los fármacos , Adulto Joven
12.
Methods Mol Biol ; 2185: 3-23, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33165839

RESUMEN

Classifying the hematological malignancies by assigning cells to their normal counterpart and describing the nature of disease progression are entirely reliant on an accurate picture for the development of the multifarious types of blood and immune cells. In recent years, our understanding of the complex relationships between the various hematopoietic stem cell-derived cell lineages has undergone substantial revision. There has been similar progress in how we describe the nature of the "target" cells that genetic insults transform to give rise to the hematological malignancies. Here I describe how both longstanding and new information has influenced classifying, for diagnosis, the hematological malignancies.


Asunto(s)
Leucemia/sangre , Leucemia/clasificación , Leucemia/inmunología , Leucemia/patología , Animales , Humanos
13.
Methods Mol Biol ; 2185: 25-37, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33165840

RESUMEN

Only 10 years ago, the existence of cancer stem cells (CSCs) was still hotly debated. Even today, when their presence in most tumor types has been clearly demonstrated, all the consequences of their existence are far from being realized neither in the clinic nor, very often, in basic and translational cancer research. The existence of CSCs supposes a true change of paradigm in our understanding of cancer, but it will only have a real impact when we will properly assimilate its implications and apply these insights to both cancer research and cancer treatment. In this primer to the topic of leukemia stem cells (LSCs) our aim is to highlight with broad brushstrokes the most relevant of their properties, how these characteristics led to their identification, and the implications that the existence of LSCs has for the research and fight against leukemia.


Asunto(s)
Leucemia , Células Madre Neoplásicas , Animales , Humanos , Leucemia/metabolismo , Leucemia/patología , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología
14.
Methods Mol Biol ; 2185: 51-63, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33165842

RESUMEN

Flow cytometry has been widely used in basic and clinical research for analysis of a variety of normal and malignant cells. Hematopoietic stem cells (HSCs) and leukemic stem cells (LSCs) can be highly purified by flow cytometry. Isolated HSCs and LSCs can be functionally identified by transplantation assays and can also be studied at the molecular level. Here we describe the flow cytometry methods for analysis and isolation of mouse HSCs and LSCs.


Asunto(s)
Citometría de Flujo , Células Madre Hematopoyéticas , Leucemia , Células Madre Neoplásicas , Animales , Células Madre Hematopoyéticas/metabolismo , Células Madre Hematopoyéticas/patología , Leucemia/metabolismo , Leucemia/patología , Ratones , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología
15.
Methods Mol Biol ; 2185: 113-134, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33165846

RESUMEN

Recurrent chromosomal translocations define genetic subtypes of childhood leukemia and present the first hit that generates an expanded clone of preleukemic cells in the bone marrow. Most commonly, reverse transcriptase PCR is used to detect these translocations on RNA level. This technique has severe drawbacks, including sensitivity to contamination and instability of RNA. Here, we describe the genomic inverse PCR for exploration of ligated breakpoints (GIPFEL) that overcomes these pitfalls.


Asunto(s)
Células de la Médula Ósea , Leucemia , Lesiones Precancerosas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células de la Médula Ósea/metabolismo , Células de la Médula Ósea/patología , Femenino , Humanos , Recién Nacido , Leucemia/genética , Leucemia/metabolismo , Leucemia/patología , Masculino , Lesiones Precancerosas/genética , Lesiones Precancerosas/metabolismo , Lesiones Precancerosas/patología
16.
Methods Mol Biol ; 2185: 361-372, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33165860

RESUMEN

Leukemia is a clonal malignant disease originated in a single cell and characterized by the accumulation of abnormal lymphoid cells. The nature of the leukemic stem cell (LSC) has been a subject of continuing discussion, given the fact that human disease is diagnosed at late stages and cannot be monitored during its natural evolution from its cell of origin. Animal models provide a means to determine the leukemic initiating cell and the causes of malignancy, and to develop new treatments. Recent findings in mice have shown that cancer stem cells can initially arise through a reprogramming-like mechanism when the oncogene expression is targeted to the mouse stem cell compartment (Garcia-Ramirez et al., EMBO J 37(14):298783, 2018; Martin-Lorenzo et al., Cancer Res 78 (10):2669-2679, 2018; Perez-Caro et al., EMBO J 28(1):8-20, 2009; Rodriguez-Hernandez et al., Cancer Res 77(16):4365-4377, 2017). If leukemia arises through reprogramming processes, then perhaps many of the oncogenes that initiate tumor formation might be dispensable for tumor progression and maintenance. Leukemia will be modeled in the mice only if we are able to target the right cancer-initiating cell with a precise given oncogene. In the last years, some examples have already started to appear in the literature showing that targeting oncogene expression to the stem cell compartment in model mice might be the correct way of reproducing the genotype-phenotype correlations found in human leukemias (Garcia-Ramirez et al., EMBO J 37(14):298783, 2018; Martin-Lorenzo et al., Cancer Res 78 (10):2669-2679, 2018; Perez-Caro et al., EMBO J 28(1):8-20, 2009; Rodriguez-Hernandez et al., Cancer Res 77(16):4365-4377, 2017). This chapter addresses how to generate LSCs by transgenesis in a way that makes the resulting animal models valuable tools to reproduce and understand leukemogenesis, and for the development of therapeutic applications like drug discovery or biomarker identification.


Asunto(s)
Transformación Celular Neoplásica , Técnicas de Reprogramación Celular , Regulación Leucémica de la Expresión Génica , Leucemia , Células Madre Neoplásicas/metabolismo , Oncogenes , Animales , Humanos , Leucemia/genética , Leucemia/metabolismo , Leucemia/patología , Ratones , Células Madre Neoplásicas/patología
17.
Am J Clin Pathol ; 155(2): 179-210, 2021 02 04.
Artículo en Inglés | MEDLINE | ID: mdl-33367563

RESUMEN

OBJECTIVES: To report the findings of the 2019 Society for Hematopathology/European Association for Haematopathology Workshop within the categories of reactive eosinophilia, hypereosinophilic syndrome (HES), germline disorders with eosinophilia (GDE), and myeloid and lymphoid neoplasms associated with eosinophilia (excluding entities covered by other studies in this series). METHODS: The workshop panel reviewed 109 cases, assigned consensus diagnosis, and created diagnosis-specific sessions. RESULTS: The most frequent diagnosis was reactive eosinophilia (35), followed by acute leukemia (24). Myeloproliferative neoplasms (MPNs) received 17 submissions, including chronic eosinophilic leukemia, not otherwise specified (CEL, NOS). Myelodysplastic syndrome (MDS), MDS/MPN, and therapy-related myeloid neoplasms received 11, while GDE and HES received 12 and 11 submissions, respectively. CONCLUSIONS: Hypereosinophilia and HES are defined by specific clinical and laboratory criteria. Eosinophilia is commonly reactive. An acute leukemic onset with eosinophilia may suggest core-binding factor acute myeloid leukemia, blast phase of chronic myeloid leukemia, BCR-ABL1-positive leukemia, or t(5;14) B-lymphoblastic leukemia. Eosinophilia is rare in MDS but common in MDS/MPN. CEL, NOS is a clinically aggressive MPN with eosinophilia as the dominant feature. Bone marrow morphology and cytogenetic and/or molecular clonality may distinguish CEL from HES. Molecular testing helps to better subclassify myeloid neoplasms with eosinophilia and to identify patients for targeted treatments.


Asunto(s)
Eosinofilia , Neoplasias Hematológicas , Síndrome Hipereosinofílico , Leucemia Linfoide , Diagnóstico Diferencial , Eosinofilia/diagnóstico , Eosinofilia/etiología , Eosinofilia/patología , Femenino , Proteínas de Fusión bcr-abl/metabolismo , Predisposición Genética a la Enfermedad , Células Germinativas/patología , Neoplasias Hematológicas/diagnóstico , Neoplasias Hematológicas/patología , Técnicas Histológicas , Humanos , Síndrome Hipereosinofílico/diagnóstico , Síndrome Hipereosinofílico/patología , Leucemia/diagnóstico , Leucemia/patología , Leucemia Linfoide/diagnóstico , Leucemia Linfoide/patología , Leucemia Mielógena Crónica BCR-ABL Positiva/diagnóstico , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Leucemia Mieloide de Fase Acelerada/diagnóstico , Leucemia Mieloide de Fase Acelerada/patología , Masculino , Síndromes Mielodisplásicos/diagnóstico , Síndromes Mielodisplásicos/patología , Patología Molecular , Leucemia-Linfoma Linfoblástico de Células T Precursoras/diagnóstico , Leucemia-Linfoma Linfoblástico de Células T Precursoras/patología
18.
J Ethnopharmacol ; 266: 113458, 2021 Feb 10.
Artículo en Inglés | MEDLINE | ID: mdl-33039632

RESUMEN

ETHNOPHARMACOLOGICAL RELEVANCE: Fumaria officinalis (Fumariaceae) is recorded in the Kurdish ethnobotany for various health problems. AIM OF THE STUDY: In this study, the cytotoxic activity of F. officinalis extracts on two leukemia and nine multiple myeloma (MM) cell lines was investigated. MATERIALS AND METHODS: The cytotoxic and ferroptotic activity were examined by resazurin reduction assay. Flow cytometry, immunoblotting assay and fluorescence microscopy were used to measure cell cycle distribution, apoptosis, induction of reactive oxygen species (ROS), loss integrity of mitochondrial membrane potential (MMP) and autophagy. LC-ESI/MS was used to identify chemical constituents present in F. officinalis. RESULTS: Chloroform (CF) and ethyl acetate (EF) fractions showed drastic cytotoxic effect on CCRF-CEM and CEM/ADR 5000 cells. NCI-H929 cell line exhibited higher sensitivity against CF, while EF demonstrated its higher cytotoxicity on OPM-2 cells with IC50 value 14.80 ± 1.70 and 28.13 ± 1.38 µg/mL respectively. Flow cytometric and morphological studies confirmed that CF and EF induced apoptosis in NCI-H929 cells by loss of MMP, generation of ROS and obvious morphological variations. In DNA histograms, up to 50% of the cells were accumulated by CF and 44% by EF in the sub-G0/G1 phase following 72 h treatment. EF induced autophagic cell death, while CF stimulated iron-dependent cell death. Moreover, two isoquinoline alkaloids and four flavonoids were identified in the active fractions. CONCLUSION: To our knowledge, this is the first report demonstrating the cytotoxicity of F. officinalis extracts in MM cell lines. CF and EF fractions inhibited MM cell proliferation through various modes of actions.


Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Fumaria/química , Leucemia/tratamiento farmacológico , Mieloma Múltiple/tratamiento farmacológico , Extractos Vegetales/farmacología , Antineoplásicos Fitogénicos/administración & dosificación , Antineoplásicos Fitogénicos/aislamiento & purificación , Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Línea Celular Tumoral , Humanos , Concentración 50 Inhibidora , Leucemia/patología , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mieloma Múltiple/patología , Extractos Vegetales/administración & dosificación , Especies Reactivas de Oxígeno/metabolismo
19.
Phytomedicine ; 81: 153409, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33341310

RESUMEN

BACKGROUND: Sesquiterpene lactones having α-methylene-γ-lactone moiety are promising natural metabolites showing various biological activity. One of the major metabolites isolated from Pulicaria undulata, 2α-hydroxyalantolactone (PU-1), has not been investigated in detail yet. Multidrug resistance (MDR) represents a major obstacle for cancer chemotherapy and the capability of novel natural products to overcoming MDR is of great interest. PURPOSE: Exploring the molecular modes of action for potent natural product metabolites. METHODS: The resazurin reduction assay was employed to evaluate the cytotoxicity of PU-1 on sensitive and their corresponding drug-resistant cell lines (overexpressing P-glycoprotein, BCRP, ABCB5, ΔEGFR, or TP53 knockout). Gene expression profiling was performed by transcriptome-wide mRNA microarray in the human CCRF-CEM leukemic cells after treatment with PU-1. The top significantly up- or down-regulated genes were identified by Chipster program and analyzed using Ingenuity Pathway Analysis (IPA) software. Finally, flow cytometry and Western blotting were performed for cell cycle analyses and apoptosis detection. RESULTS: The sesquiterpene lactone, PU-1, showed potent cytotoxicity towards the drug-sensitive and -resistant cell lines. Transcriptome-wide mRNA expression profiling and pathway analysis pointed to genes involved in DNA damage response and G2/M cell cycle arrest. G2/M arrest was verified by flow cytometry and further confirmed by the upregulation of p21 and downregulation of p-CDC25C expression in Western blotting. Moreover, the suggested DNA damage checkpoint regulation was confirmed by immunofluorescence and Western blotting by upregulation of pS345 Chk1, p-H3 and γ-H2AX. Furthermore, PU-1 inhibited PI3K/AKT pathway, which is involved in signaling DNA damage and G2/M arrest. Cells ultimately induced apoptosis upon PU-1 treatment. CONCLUSIONS: PU-1 is a potent natural product inhibiting otherwise drug-resistant human tumor cell growth through DNA damage, G2/M cell cycle arrest and apoptosis.


Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Resistencia a Antineoplásicos/efectos de los fármacos , Leucemia/tratamiento farmacológico , Pulicaria/química , Sesquiterpenos/farmacología , Antineoplásicos Fitogénicos/química , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Daño del ADN/efectos de los fármacos , Resistencia a Antineoplásicos/fisiología , Puntos de Control de la Fase G2 del Ciclo Celular/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Leucemia/patología , Fosfatidilinositol 3-Quinasas/metabolismo , Inhibidores de las Quinasa Fosfoinosítidos-3/farmacología , Sesquiterpenos/química
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA
...