RESUMO
The S-doped mesoporous nanocomposite (S-TNT) of HTiNbO5 nanosheets (NSs) and anatase TiO2 nanoparticles (NPs) with exposed {101} facets has been successfully synthesized by first mixing freeze-dried HTiNbO5 NSs with titanium isopropoxide and then calcination with thiourea in air. The exposed anatase {101} facets can act as a possible reservoir of the photogenerated electrons, yielding a highly reactive surface for the reduction of O2 to O2Ë(-). The partial substitution of Ti(4+) by S(6+) in the lattice of TiO2 NPs leads to a charge imbalance in S-TNT and the formation of Ti-O-S bonds. As a result, the formed cationic S-TNT favours adsorption of hydroxide ions (OH(-)(ads)) and thus captures the photo-induced holes to form hydroxyl radicals (ËOH). Moreover, with the formation of Ti-O-S bonds, partial electrons can be transferred from S to O atoms and hence the electron-deficient S atoms might capture photo-induced electrons. The surface-adsorbed SO4(2-) could also act as an efficient electron trapping center to promote the separation of charge carriers. In addition, the Ti(3+) species due to the removal of oxygen atoms during calcination and the associated oxygen vacancy defects on the surface of S-TNT could act as hole and electron scavengers, respectively. Besides, the closely contacted interface is formed between HTiNbO5 NSs and anatase TiO2 NPs due to the common features of TiO6 octahedra in two components, resulting in a nanoscale heterojunction structure to speed up the separation rate of photogenerated charge carriers. The formation of a nano-heterojunction and the incorporation of Ti(3+) and S dopants give rise to the visible and near-infrared light response of S-TNT. The combined effects greatly retard the charge recombination and improve the photocatalytic activity for the degradation of rhodamine B (RhB) and phenol solution under visible light irradiation. The corresponding photocatalytic mechanism was investigated via the active species capture experiments. The present work may provide an insight into the fabrication of delicate composite photocatalysts with excellent performance.
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A novel asymmetric supercapacitor device in an aqueous electrolyte is fabricated using a vanadyl phosphate/carbon nanocomposite as the positive electrode and a polypyrrole-derived carbon nanowire as the negative electrode. The vanadyl phosphate/carbon nanocomposites are synthesized by a simple two-step approach in which layered VOPO4·2H2O is first intercalated by dodecylamine and then annealed at high temperature, leading to the in situ carbonization of the intercalated dodecylamine. It is found that the sample in which the incorporated carbon with a high degree of graphitization exhibits a high specific capacitance of 469 F g-1 at a current density of 1 A g-1 and excellent rate performance (retained 77% capacitance at 10 A g-1). A polypyrrole-derived carbon nanowire is synthesized by the direct carbonization of nanowire-shaped polypyrrole, revealing a rough surface of nanowire-like frameworks and good electrochemical behavior. Taking advantage of both positive and negative materials, the assembled asymmetric supercapacitor device exhibits a high energy density of 30.6 W h kg-1 at a high power density of 813 W kg-1 in a wide voltage region of 0-1.6 V, as well as a good electrochemical stability (84.3% capacitance retention after 5000 cycles). The present work can shed light on the fabrication of novel asymmetric supercapacitors with high-performance.
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OBJECTIVE: To quantitatively detect the methylation of E-cadherin gene 5'-CpG islands in acute leukemia by microarray-based DNA analysis and to briefly discuss the role of microarry for detection of methylation in tumors. METHODS: Bisulfite-modified DNA was used as a template for PCR amplification, resulting in conversion of unmethylated cytosine, but not methylated cytosine, into thymine within CpG islands of interest. Five sets of oligonucleotide probes were designed to fabricate a DNA microarray to detect the methylation changes of E-cadherin gene CpG islands in acute leukemia. By drawing a standard curve to assess the levels of changes in methylation detected in the examined samples. RESULTS: Microarray assay was successfully used to quantitatively detect methylation changes of E-cadherin gene in 5 acute leukemia samples. Varying degree of methylation was detected in five regions and the hypermethylation region was the same. The result was validated by gene sequencing. CONCLUSION: Microarray assay may be applied as an useful tool for mapping methylation changes in multiple CpG loci and for leukemia research. It is more time-saving and labor-saving than gene sequencing and can be used to quantitatively detect changes in methylation with high throughput.
Assuntos
Caderinas/genética , Metilação de DNA , Leucemia Mieloide Aguda/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Regiões Promotoras Genéticas , Sequência de Bases , Ilhas de CpG/genética , Humanos , Dados de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos/métodosRESUMO
Catalytic behavior of Co-based/zeolites catalysts was investigated in NOx reduction by CH4. Optimization of promoter and support was investigated by catalytic tests, and the relationship between catalytic activity and catalyst structure was illustrated by catalyst characterization. Co-Fe/SAPO-34 exhibited the highest activity among various Co-base/zeolites catalysts. The maximum conversion of NOx with 52.7% was obtained on Co-Fe/SAPO-34 at 450â. The inhibition of activity of Fe/zeolites became severe in the presence of SO2, CO2, and H2O. CO2 exerted virtually no effect on the SCR activity of Co-Fe/zeolites. The inhibition of NOx conversion by H2O was reversible for Co-Fe/zeolites catalysts. Cobalt species were mainly present in CoO and Co(OH)2 states in Co-Fe/SAPO-34. Co3O4and Co(OH)2 were the main cobalt species of Co-Fe/ZSM-5, while CoO, CoAl2O4 and Co3O4 might be present in Co-Fe/Beta. The ratio of Fe2+/Fe3+ in the surface layer of Co-Fe/zeolites decreased in the order of Co-Fe/ZSM-5(3.98) > Co-Fe/SAPO-34(0.52) > Co-Fe/Beta(0.43). The active states of cobalt species and suitable ratio of Fe2+/Fe3+ were important for the activity of Co-Fe/zeolites in CH4-SCR. CH4-SCR over Co-Fe/zeolite catalysts started with the adsorption of NO and CH4 on Brønsted acid sites of the zeolite to produce NO+and carbon-containing species(-C=O and -COO) in the presence of oxygen, respectively. Subsequently, the important intermediates of nitrate species were generated from NO+ at the active sites. Finally, nitrate species reacted with carbon-containing species to form N2 and CO2.
RESUMO
In neurons, a highly regulated microtubule cytoskeleton is essential for many cellular functions. These include axonal transport, regional specialization and synaptic function. Given the critical roles of microtubule-associated proteins (MAPs) in maintaining and regulating microtubule stability and dynamics, we sought to understand how this regulation is achieved. Here, we identify a novel LisH/WD40 repeat protein, tentatively named nemitin (neuronal enriched MAP interacting protein), as a potential regulator of MAP8-associated microtubule function. Based on expression at both the mRNA and protein levels, nemitin is enriched in the nervous system. Its protein expression is detected as early as embryonic day 11 and continues through adulthood. Interestingly, when expressed in non-neuronal cells, nemitin displays a diffuse pattern with puncta, although at the ultrastructural level it localizes along the microtubule network in vivo in sciatic nerves. These results suggest that the association of nemitin to microtubules may require an intermediary protein. Indeed, co-expression of nemitin with microtubule-associated protein 8 (MAP8) results in nemitin losing its diffuse pattern, instead decorating microtubules uniformly along with MAP8. Together, these results imply that nemitin may play an important role in regulating the neuronal cytoskeleton through an interaction with MAP8.
Assuntos
Peptídeos e Proteínas de Sinalização Intracelular/fisiologia , Proteínas Associadas aos Microtúbulos/fisiologia , Microtúbulos/metabolismo , Sequência de Aminoácidos , Animais , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Camundongos , Proteínas dos Microfilamentos/química , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Microtúbulos/efeitos dos fármacos , Dados de Sequência Molecular , Proteínas do Tecido Nervoso , Neurônios/metabolismoRESUMO
While the potential impact of magnetic nanoparticles (MNPs) has been widely explored in almost all medical fields, including cardiology, one question remains; that is whether MNPs interfere with cardiac physiological processes such as the expression and function of ion channels, especially in vivo. KCNQ(1) channels are richly expressed in cardiac myocytes and are critical to the repolarization of cardiac myocytes. In this study, we evaluated the effects of Fe(3)O(4)-magnetic nanoparticles (MNPs-Fe(3)O(4)) on the expression of KCNQ(1) in cardiac muscle of mice at rest and at different times following a single bout of swimming (SBS). Firstly, we demonstrated that the expression levels of KCNQ(1) channels are significantly up-regulated in mice following a SBS by means of reverse transcription polymerase chain reaction (RT-PCR) and western-blot. After treating mice with normal saline or pure MNPs-Fe(3)O(4) separately, we studied the potential effect of MNPs-Fe(3)O(4) on the expression profile of KCNQ(1) in mouse cardiac muscle following a SBS. A SBS increased the transcription of KCNQ(1) at 3 hours post exercise (3PE) 164% +/- 24% and at 12 hours post exercise (12PE) by 159% +/- 23% (P < 0.05), and up-regulated KCNQ(1) protein 161% +/- 27% at 12PE (P < 0.05) in saline mice. In MNPs-Fe(3)O(4) mice, KCNQ(1) mRNA increased by 151% +/- 14% and 147% +/- 12% at 3 and 12 PE, respectively (P <0.05). Meanwhile, an increase of 152% +/- 14% in KCNQ(1) protein was also detected at by 12PE. These results indicated that the administration of MNPs-Fe(3)O(4) did not cause any apparent effects on the expression profile of KCNQ(1) in rested or exercised mice cardiac muscle. Our studies suggest a novel path of KCNQ(1) current adaptations in the heart during physical exercise and in addition provide some useful information for the biomedical application of MNPs which are imperative to advance nanomedicine.
Assuntos
Compostos Férricos/administração & dosagem , Coração/fisiologia , Canal de Potássio KCNQ1/metabolismo , Nanopartículas/administração & dosagem , Esforço Físico/fisiologia , Animais , Campos Eletromagnéticos , Compostos Férricos/química , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/fisiologia , Coração/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Nanopartículas/química , Nanopartículas/ultraestrutura , Tamanho da Partícula , Esforço Físico/efeitos dos fármacosRESUMO
The Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway is active in both normal hematopoiesis and hematological malignancies. Moreover, Janus kinase-2 (JAK2) is the key hematopoietic kinase, and mutations together with single nucleotide polymorphisms (SNPs) of JAK2 have been thoroughly evaluated. In this study, we aimed to investigate whether the synonymous genetic polymorphism A830G in the JAK2 gene is associated with the treatment outcomes of Ara-C-based chemotherapy regimens in patients with acute myeloid leukemia (AML). A total of 152 patients with AML in a Chinese population were enrolled in our study. Peripheral blood samples drawn at the time of diagnosis were analyzed for the presence of JAK2 A830G by matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS). The results showed that frequencies of the AA genotype were higher in the group 40-60 years old, higher white blood cell (WBC) patient group, homoharringtonine and Ara-C (HA) regimen group, and good therapy response group, and patients with the GG genotype were significantly associated with a higher rate of early death. We conclude that the AA and GG genotypes of JAK2 A830G might be important markers for therapy outcomes of patients with AML in a Chinese population.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Janus Quinase 2/genética , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/genética , Polimorfismo de Nucleotídeo Único , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Povo Asiático/genética , China , Citarabina/administração & dosagem , Feminino , Frequência do Gene , Genótipo , Humanos , Leucemia Mieloide Aguda/etnologia , Masculino , Pessoa de Meia-Idade , Indução de Remissão , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Resultado do Tratamento , Adulto JovemRESUMO
This study was aimed to explore the potential therapy of Gambogic acid (GA) combined with magnetic nanoparticle of Fe3O4 (Fe3O4-MNP) on leukemia. The proliferation of U937 cells and the cytotoxicity were evaluated by MTT assay. Cell apoptosis was observed and analyzed by microscopy and flow cytometry respectively. The expressions of gene and protein were detected by quantitative real-time polymerase chain reaction and Western blot respectively. The results showed that GA enhanced the cytotoxicity for U937 cells in dose- and time-dependent manners. The Fe3O4-MNP itself had not cytotoxicity, but could enhance the inhibitory effect of GA on proliferation of U937 cells. The apoptotic rate of U937 cells induced by combination of GA with Fe3O4-MNP was higher than that by GA alone. The typical apoptotic features of cells treated with GA and Fe3O4-MNP were observed. The expression levels of caspase-3 and bax after co-treatment of GA and Fe3O4-MNP were higher than that exposed to GA or Fe3O4-MNP alone, but the expressions of bcl-2, NF-kappaB and survivin were down-regulated. It is concluded that Fe3O4-MNP can promote GA-induced apoptosis in U937 cells, and the combination of GA with Fe3O4-MNP may be a safer and less toxic new therapy for leukemia.
Assuntos
Apoptose/efeitos dos fármacos , Compostos de Ferro/administração & dosagem , Compostos de Ferro/farmacologia , Xantonas/farmacologia , Humanos , Magnetismo , Nanopartículas , Células U937RESUMO
In many instances, multidrug resistance (MDR) is mediated by increasing the expression at the cell surface of the MDR1 gene product, P-glycoprotein (P-gp), a 170-kD energy-dependent efflux pump. The aim of this study was to investigate the potential benefit of combination therapy with magnetic Fe(3)O(4) nanoparticle [MNP (Fe(3)O(4))] and MDR1 shRNA expression vector in K562/A02 cells. For stable reversal of "classical" MDR by short hairpin RNA (shRNA) aiming directly at the target sequence (3491-3509, 1539-1557, and 3103-3121 nucleotide) of MDR1 mRNA. PGC silencer-U6-neo-GFP-shRNA/MDR1 called PGY1-1, PGY1-2, and PGY1-3 were constructed and transfected into K562/A02 cells by lipofectamine 2000. After transfected and incubated with or without MNP (Fe(3)O(4)) for 48 hours, the transcription of MDR1 mRNA and the expression of P-gp were detected by quantitative real-time PCR and Western-blot assay respectively. Meanwhile intracellular concentration of DNR in K562/A02 cells was detected by flow cytometry (FCM). PGC silencer-U6-neo-GFP-shRNA/MDR1 was successfully constructed, which was confirmed by sequencing and PGY1-2 had the greatest MDR1 gene inhibitory ratio. Analysis of the reversal ratio of MDR, the concentration of daunorubicin (DNR) and the transcription of MDR1 gene and expression of P-gp in K562/A02 showed that combination of DNR with either MNP (Fe(3)O(4)) or PGY1-2 exerted a potent cytotoxic effect on K562/A02 cells, while combination of MNP (Fe(3)O(4)) and PGY1-2 could synergistically reverse multidrug resistance. Thus our in vitro data strongly suggested that a combination of MNP (Fe(3)O(4)) and shRNA expression vector might be a more sufficient and less toxic anti-MDR method on leukemia.
Assuntos
Daunorrubicina/administração & dosagem , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Resistência a Múltiplos Medicamentos/genética , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/genética , Genes MDR , Nanopartículas de Magnetita/administração & dosagem , RNA Interferente Pequeno/administração & dosagem , RNA Interferente Pequeno/genética , Subfamília B de Transportador de Cassetes de Ligação de ATP , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/antagonistas & inibidores , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Antibióticos Antineoplásicos/administração & dosagem , Sequência de Bases , Primers do DNA/genética , Doxorrubicina/administração & dosagem , Vetores Genéticos , Humanos , Células K562 , Dados de Sequência Molecular , Nanomedicina , TransfecçãoRESUMO
Apoptosis is a common pathway that finally mediated the killing functions of anticancer drugs, which is an important cause of multidrug resistance (MDR). The aim of this study was to investigate the potential benefit of combination therapy with magnetic nanoparticle of Fe(3)O(4) (MNP(Fe(3)O(4))) and 5-bromotetrandrin (BrTet). Analysis of the apoptosis percentage showed that combination of daunorubicin (DNR) with either MNP(Fe(3)O(4)) or BrTet exerted a potent cytotoxic effect on K562/A02 cells, while MNP(Fe(3)O(4)) and BrTet cotreatment can synergistically enhance DNR-induced apoptosis. Importantly, we confirmed that the distinct synergism effect of that composite on reverse multidrug resistance may owe to the regulation of various proliferative and antiapoptotic gene products, including P53 and caspase-3. Thus our in vitro data strongly suggests a potential clinical application of MNP(Fe(3)O(4)) and BrTet combination on CML.
Assuntos
Apoptose/efeitos dos fármacos , Benzilisoquinolinas/química , Daunorrubicina/administração & dosagem , Portadores de Fármacos/química , Compostos Férricos/química , Leucemia/fisiopatologia , Nanopartículas/química , Antibióticos Antineoplásicos/administração & dosagem , Antibióticos Antineoplásicos/química , Benzilisoquinolinas/efeitos da radiação , Daunorrubicina/química , Portadores de Fármacos/efeitos da radiação , Compostos Férricos/efeitos da radiação , Humanos , Células K562 , Nanopartículas/administração & dosagem , Nanopartículas/efeitos da radiaçãoRESUMO
In this paper we establish the xenograft leukemia model with stable multidrug resistance in nude mice and to investigate the reversal effect of 5-bromotetrandrine (5-BrTet) and magnetic nanoparticle of Fe(3)O(4) (MNP-Fe(3)O(4)) combined with daunorubicin (DNR) in vivo. Two subclones of K562 and K562/A02 cells were inoculated subcutaneously into the back of athymic nude mice (1 x 10(7) cells/each) respectively to establish leukemia xenograft models. Drug-resistant and sensitive tumor-bearing nude mice were assigned randomly into five groups which were treated with normal saline; DNR; NP-Fe(3)O(4) combined with DNR; 5-BrTet combined with DNR; 5-BrTet and MNP-Fe(3)O(4) combined with DNR, respectively. The incidence of formation, growth characteristics, weight, and volume of tumors were observed. The histopathologic examination of tumors and organs were detected. For resistant tumors, the protein levels of Bcl-2, and BAX were detected by Western blot. Bcl-2, BAX, and caspase-3 genes were also detected. For K562/A02 cells xenograft tumors, 5-BrTet and MNP-Fe(3)O(4) combined with DNR significantly suppressed growth of tumor. A histopathologic examination of tumors clearly showed necrosis of the tumors. Application of 5-BrTet and MNP-Fe(3)O(4) inhibited the expression of Bcl-2 protein and upregulated the expression of BAX and caspase-3 proteins in K562/A02 cells xenograft tumor. It is concluded that 5-BrTet and MNP-Fe(3)O(4) combined with DNR had a significant tumor-suppressing effect on a MDR leukemia cells xenograft model.
Assuntos
Apoptose/efeitos dos fármacos , Benzilisoquinolinas/química , Daunorrubicina/administração & dosagem , Portadores de Fármacos/química , Compostos Férricos/química , Leucemia/tratamento farmacológico , Nanopartículas/química , Animais , Antibióticos Antineoplásicos/administração & dosagem , Antibióticos Antineoplásicos/química , Benzilisoquinolinas/efeitos da radiação , Daunorrubicina/química , Portadores de Fármacos/efeitos da radiação , Resistência a Múltiplos Medicamentos , Compostos Férricos/efeitos da radiação , Humanos , Células K562 , Leucemia/patologia , Camundongos , Camundongos Nus , Nanopartículas/administração & dosagem , Nanopartículas/efeitos da radiação , Resultado do TratamentoRESUMO
Multidrug resistance (MDR) is a major obstacle to cancer chemotherapy. We evaluated the effect of daunorubicin (DNR)-loaded magnetic nanoparticles of Fe3O4 (MNPs-Fe3O4) on K562-n/VCR cells in vivo. K562-n and its MDR counterpart K562-n/VCR cell were inoculated into nude mice subcutaneously. The mice were randomly divided into four groups: group A received normal saline, group B received DNR, group C received MNPs-Fe3O4, and group D received DNR-loaded MNPs-Fe3O4. For K562-n/VCR tumor, the weight was markedly lower in group D than that in groups A, B, and C. The transcriptions of Mdr-1 and Bcl-2 gene were significantly lower in group D than those in groups A, B, and C. The expression of Bcl-2 was lower in group D than those in groups A, B, and C, but there was no difference in the expression of P-glycoprotein. The transcriptions and expressions of Bax and caspase-3 in group D were increased significantly when compared with groups A, B, and C. In conclusion, DNR-loaded MNPs-Fe3O4 can overcome MDR in vivo.
Assuntos
Antibióticos Antineoplásicos/administração & dosagem , Daunorrubicina/administração & dosagem , Óxido Ferroso-Férrico/administração & dosagem , Nanopartículas Metálicas/administração & dosagem , Animais , Sequência de Bases , Primers do DNA/genética , Sistemas de Liberação de Medicamentos , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Feminino , Expressão Gênica/efeitos dos fármacos , Humanos , Células K562 , Leucemia Experimental/tratamento farmacológico , Leucemia Experimental/genética , Leucemia Experimental/patologia , Magnetismo , Camundongos , Camundongos Nus , Nanomedicina , Transplante de Neoplasias , RNA Neoplásico/genética , Transplante HeterólogoRESUMO
This study aims to evaluate the multidrug resistance (MDR) reversal activity by magnetic nanoparticles of Fe3O4 (MNPs-Fe3O4) and 5-bromotetrandrine (BrTet) MDR cell line K562/A02 solitarily or symphysially. The proliferation of K562 and K562/A02 cells and the cytotoxicity on peripheral blood mononuclear cells (PMBCs) were evaluated by MTT assay. Cellular accumulation of daunorubicin (DNR) was analyzed by flow cytometry. Real-time polymerase chain reaction and Western blotting analyses were performed to examine the mRNA and protein levels of mdr1, respectively. The results showed that the combination of MNPs-Fe3O4 and BrTet with effective concentrations significantly increased cytotoxicity against MDR cell line K562/A02. Both BrTet and MNPs-Fe3O4 increased the intracellular DNR accumulation in the K562/A02 cell line, and downregulated the level of mdr1 gene and expression of P-glycoprotein. Furthermore, the combination did not have significant cytotoxicity in PMBCs. We propose that MNPs-Fe3O4 conjugated with DNR and BrTet probably have synergetic effects on MDR reversal.
Assuntos
Benzilisoquinolinas/administração & dosagem , Óxido Ferroso-Férrico/administração & dosagem , Nanopartículas Metálicas/administração & dosagem , Subfamília B de Transportador de Cassetes de Ligação de ATP , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/antagonistas & inibidores , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Antibióticos Antineoplásicos/administração & dosagem , Antibióticos Antineoplásicos/farmacocinética , Sequência de Bases , Sobrevivência Celular/efeitos dos fármacos , Daunorrubicina/administração & dosagem , Daunorrubicina/farmacocinética , Regulação para Baixo/efeitos dos fármacos , Resistência a Múltiplos Medicamentos/genética , Resistencia a Medicamentos Antineoplásicos/genética , Sinergismo Farmacológico , Genes MDR/efeitos dos fármacos , Humanos , Células K562 , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Leucemia Mielogênica Crônica BCR-ABL Positiva/genética , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Magnetismo , Nanomedicina , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Neoplásico/genética , RNA Neoplásico/metabolismoRESUMO
The present study was aimed to evaluate the MDR reversal activity of bromotetrandrine (BrTet) in vitro and in vivo. The inhibitory effects of adriamycin (ADM) used alone or in combination with BrTet or Tet on the proliferation of K562 and K562/A02 cells were evaluated by MTT assay. The ADM accumulation and the protein levels of P-glycoprotein (P-gp) were detected by flow cytometry. The mRNA levels of P-gp were determined by RT-PCR. The in vivo effect of BrTet and Tet was investigated by using nude mice grafted with sensitive human leukemia cell line K562 and MDR cell line K562/A02. The results showed that BrTet at 0.25, 0.5 and 1 micromol/L reversed the resistance to ADM in MDR K562/A02 cells in a dose-dependent manner. Flow cytometry suggested that BrTet significantly increased the intracellular accumulation of ADM in K562/A02 cells in a dose-dependent manner. BrTet also inhibited the overexpression of P-gp in K562/A02 cells, and down-regulated mdr1 expression. In nude mice bearing K562 xenografts on the left flank and K562/A02 xenografts on the right flank, intraperitoneal injection of 10 mg/kg BrTet significantly enhanced the antitumor activity of ADM against K562/A02 xenografts with inhibitory rates of 26.1%, while ADM alone inhibited the growth of K562/A02 xenografts only by 5.8%. No enhancement effect by BrTet was seen in K562 xenografts. It is concluded that BrTet shows significant MDR reversal activity in vitro and in vivo. Its activity may be related to the inhibition of P-gp overexpression and the increase intracellular accumulation of anticancer drugs. BrTet may be a promising-MDR modulator for eventual assessment in the clinic.
Assuntos
Benzilisoquinolinas/farmacologia , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Subfamília B de Transportador de Cassetes de Ligação de ATP , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Animais , Resistência a Múltiplos Medicamentos/genética , Resistencia a Medicamentos Antineoplásicos/genética , Feminino , Humanos , Células K562 , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Drug resistance is a primary hindrance for efficiency of chemotherapy. To investigate whether Fe3O4-magnetic nanoparticles (Fe3O4-MNPs) loaded with adriamycin (ADM) and tetrandrine (Tet) would play a synergetic reverse role in multidrug resistant cell, we prepared the drug-loaded nanoparticles by mechanical absorption polymerization to act with K562 and one of its resistant cell line K562/A02. The survival of cells which were cultured with these conjugates for 48 h was observed by MTT assay. Using cells under the same condition described before, we took use of fluorescence microscope to measure fluorescence intensity of intracellular ADM at an excitation wavelength of488 nm. P-glycoprotein (P-gp) was analyzed with flow cytometer. The expression ofmdrl mRNA was measured by RT-PCR. The results showed that the growth inhibition efficacy of both the two cells increased with augmenting concentrations of Fe3O4-MNPs which were loaded with drugs. No linear correlation was found between fluorescence intensity of intracellular adriamycin and augmenting concentration of Fe3O4-MNPs. Tet could downregulate the level of mdr-1 gene and decrease the expression of P-gp. Furthermore, Tet polymerized with Fe3O4-MNPs reinforced this downregulation, causing a 100-fold more decrease in mdrl mRNA level, but did not reduce total P-gp content. Our results suggest that Fe3O4-MNPs loaded with ADM or Tet can enhance the effective accumulation of the drugs in K562/A02. We propose that Fe3O4-MNPs loaded with ADM and Tet probably have synergetic effect on reversal in multidrug resistance.
Assuntos
Benzilisoquinolinas/administração & dosagem , Sobrevivência Celular/efeitos dos fármacos , Doxorrubicina/administração & dosagem , Portadores de Fármacos/administração & dosagem , Compostos Férricos/química , Nanopartículas/administração & dosagem , Antineoplásicos/administração & dosagem , Proliferação de Células , Doxorrubicina/química , Portadores de Fármacos/química , Resistência a Múltiplos Medicamentos , Humanos , Células K562 , Magnetismo/uso terapêutico , Nanopartículas/química , Nanopartículas/ultraestrutura , Tamanho da PartículaRESUMO
OBJECTIVE: To study the effect of tetrandrine (Tet) in combination with droloxifen (DRL) on the expression of nuclear factor kappa B (NF-kappaB) in K562 and K562/A02 cell lines and its reversal mechanism. METHODS: The activation of NF-kappaB in K562 and K562/A02 cell lines and the effect of Tet or DRL alone or in combination on NF-kappaB protein expression were determined with immunocytochemistry and Western blotting respectively. RESULTS: (1) K562/A02 cells displayed higher level of NF-kappaB protein expression than K562 cells. (2) The application of Tet or DRL alone or in combination had no effect on NF-kappaB expression in K562 cells at 6 h and 12 h (P > 0.05). (3) Tet and DRL alone or in combination could significantly down-regulate NF-kappaB protein expression in nuclei of K562/A02 cells. The effect was more significant in combination than either alone. This effect was more significant at 12 h than at 6 h. CONCLUSIONS: (1) Activation of NF-kappaB may be involved in the mechanism of MDR of K562/A02 cell line. (2)Inhibition of NF-kappaB activation may be involved in the reversal of multidrug resistance in K562/A02 cells by Tet and DRL.
Assuntos
Benzilisoquinolinas/farmacologia , NF-kappa B/metabolismo , Tamoxifeno/análogos & derivados , Resistência a Múltiplos Medicamentos , Humanos , Células K562 , Tamoxifeno/farmacologiaRESUMO
In this study, we have explored the possibility of the combination of the high reactivity of nano Fe3O4 or Au nanoparticles and daunomycin, one of the most important antitumor drugs in the treatment of acute leukemia clinically, to inhibit MDR of K562/A02 cells. Initially, to determine whether the magnetic nanoparticle Fe3O4 and Au can facilitate the anticancer drug to reverse the resistance of cancer cells, we have explored the cytotoxic effect of daunomycin (DNR) with and without the magnetic nano-Fe3O4 or nano-Au on K562 and K562/A02 cells by MTT assay. Besides, the intracellular DNR concentration and apoptosis of the K562/A02 cells was further investigated by flow cytometry and confocal fluorescence microscopic studies. The MDR1 gene expression of the K562/A02 cells was also studied by RT-PCR method. Our results indicate that 5.0 x 10(-7) M nano-Fe3O4 or 2.0 x 10(-8) M nano-Au is biocompatible and can apparently raise the intracellular DNR accumulation of the K562/A02 cells and increase the apoptosis of tumor cells. Moreover, our observations illustrate that although these two kinds of nanoparticles themselves could not lower the MDRI gene expression of the K562/A02 cells, yet they could degrade the MDR1 gene level when combining with anticancer drug DNR. This raises the possibility to combine the nano-Fe3O4 or nano-Au with DNR to reverse the drug resistance of K562/A02 cells, which could offer a new strategy for the promising efficient chemotherapy of the leukemia patients.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Daunorrubicina/administração & dosagem , Compostos Férricos/administração & dosagem , Ouro/administração & dosagem , Leucemia/patologia , Leucemia/fisiopatologia , Nanopartículas/administração & dosagem , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Compostos Férricos/química , Ouro/química , Humanos , Células K562 , Nanopartículas/químicaRESUMO
This study was aimed to investigate the cytogenetic changes of MDS cell line (MUTZ-1) with chromosome 5q deletion. R-banding analysis was used to identify chromosome aberrations in MDS cell line and Vysis Spectra Vysion M-FISH was used to further characterize chromosomal complex karyotype. The results indicated that M-FISH exhibited obvious chromosomal aberrations with high frequency including translocation, insertion, breakage and rearrangement, deletion and increasement of chromosome number, the complex karyotype of MUTZ-1 was shown as 50, xx, der (1) t (1;2), ins (1;14), +der (2) t(2;19), der (3) t (3;5), der (3) (3::5::22), 5q-, der (6) t (3;6), der (7) (18::7::17), +8, +der (9) t (1;9), der (10) t (1;10), +11, +12, der (?13) (10::13::5::8), der (14) t (8;14), der (14) t (14, 15), der (15) t (15;21) x 2, +17, +18, -21, -22. It is concluded that M-FISH analysis revealed obvious changes in complex karyotype of MDS cell line MUTZ-1, and the M-FISH technique can increase accuracy of detection for chromosomal complex karyotype, and help diagnosis and prognostic evaluation of MDS.
Assuntos
Aberrações Cromossômicas , Cromossomos Humanos Par 5 , Hibridização in Situ Fluorescente , Síndromes Mielodisplásicas/genética , Pré-Escolar , Deleção Cromossômica , Feminino , Humanos , Hibridização in Situ Fluorescente/métodos , Cariotipagem , Translocação Genética , Células Tumorais CultivadasRESUMO
To explore whether the complete donor chimerism could be achieved and graft-versus-host disease could be alleviated by donor lymphocyte infusion which was sensitized by the skin of the recipient, female C57BL/6 mice (H-2(b), B6) as recipients received total body irradiation (TBI) of 5.5 Gy ((60)Co gamma-ray) on day 0 followed by allogeneic hematopoietic stem cell transplantation (allo-HSCT). The allo-grafts consisted of 2 x 10(7) peripheral hematopoietic stem cells from mobilized male BALB/c (H-2(d)) donor mice with the granulocyte colony-stimulating factor (G-CSF). Day 2 after allo-HSCT, the recipient mice were given 200 mg/kg cyclophosphamide intraperitoneally. Afterwards these recipient mice were infused 2 x 10(6) sensitized or unsensitized-donor lymphocytes at the 28 days after transplantation. The results showed that the mice receiving sensitized-donor lymphocyte infusion did not suffer from GVHD and the phenotypic character of the recipient mice (black color) converted to that of the donor mice (white color), and to become full-donor chimerism. It was found that the ratio of CD4(+)/CD8(+) T lymphocytes of them decreased at the earlier period and increased after half month, but which were also lower than that of the normal value. While various grades of acute GVHD was observed in that of the control group and the mixed-chimeras were maintained, though it increased a little, and the ratio of CD4(+)/CD8(+) T lymphocytes increased at first, then decreased to the normal level half month later. It is concluded that sensitized DLI converted mixed to complete donor chimerism without GVHD, and the rate of CD4(+)/CD8(+) has close relation to the incidence of GVHD.