RESUMEN
OBJECTIVES: The interferon regulatory factor (IRF) family of proteins are involved in tumor progression. However, the role of IRF5 in tumorigenesis remains unknown. In this study we aimed to elucidate the functions of IRF5 in the progression of hepatocellular carcinoma (HCC). METHODS: IRF5 expression in HCC was analyzed through quantitative polymerase chain reaction (qPCR), western blot, and immunohistochemistry (IHC), etc. The Cell Counting Kit 8 (CCK8) assay, anchorage-independent assay, and EdU assay were used to evaluate the role of IRF5. The molecular mechanisms were studied by analyzing the metabolites with mass spectrum and immunoprecipitation. RESULTS: IRF5 was upregulated in HCC. Interfering with IRF5 inhibited the proliferation and tumorigenic potential of HCC cells. When studying the molecular mechanism, IRF5 was found to upregulate the expression of lactate dehydrogenase A (LDHA) and promoted glycolysis. Additionally, tripartite motif containing 35 (TRIM35) interacted with IRF5, promoting its ubiquitination and degradation. In the clinically obtained HCC samples, TRIM35 was negatively correlated with the expression of IRF5. CONCLUSION: These findings reveal the oncogenic function of IRF5 in the progression of HCC by enhancing glycolysis, further supporting the potential of IRF5 as a viable target for HCC therapy.
Asunto(s)
Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/patología , Neoplasias Hepáticas/patología , Línea Celular Tumoral , Factores Reguladores del Interferón/genética , Factores Reguladores del Interferón/metabolismo , Glucólisis , Proliferación Celular , Regulación Neoplásica de la Expresión Génica , Proteínas Reguladoras de la Apoptosis/genéticaRESUMEN
OBJECTIVES: To study the correlation between CT imaging features of acceleration and deceleration brain injury and injury degree. METHODS: A total of 299 cases with acceleration and deceleration brain injury were collected and divided into acceleration brain injury group and deceleration brain injury group according to the injury mechanism. Subarachnoid hemorrhage (SAH) and Glasgow coma scale (GCS), combined with skull fracture, epidural hematoma (EDH), subdural hematoma (SDH) and brain contusion on the same and opposite sides of the stress point were selected as the screening indexes. χ2 test was used for primary screening, and binary logistic regression analysis was used for secondary screening. The indexes with the strongest correlation in acceleration and deceleration injury mechanism were selected. RESULTS: χ2 test showed that skull fracture and EDH on the same side of the stress point; EDH, SDH and brain contusion on the opposite of the stress point; SAH, GCS were correlated with acceleration and deceleration injury (P<0.05). According to binary logistic regression analysis, the odds ratio (OR) of EDH on the same side of the stress point was 2.697, the OR of brain contusion on the opposite of the stress point was 0.043 and the OR of GCS was 0.238, suggesting there was statistically significant (P<0.05). CONCLUSIONS: EDH on the same side of the stress point, brain contusion on the opposite of the stress point and GCS can be used as key indicators to distinguish acceleration and deceleration injury mechanism. In addition, skull fracture on the same side of the stress point, EDH and SDH on the opposite of the stress point and SAH were relatively weak indicators in distinguishing acceleration and deceleration injury mechanism.
Asunto(s)
Contusión Encefálica , Lesiones Encefálicas , Hematoma Epidural Craneal , Fracturas Craneales , Heridas no Penetrantes , Lesiones Encefálicas/diagnóstico por imagen , Hematoma Subdural/diagnóstico por imagen , Hematoma Subdural/etiología , Humanos , Modelos Logísticos , Fracturas Craneales/diagnóstico por imagen , Tomografía Computarizada por Rayos X , Heridas no Penetrantes/diagnóstico por imagenRESUMEN
BACKGROUND: The poly (ADP-ribose) polymerase (PARP) inhibitor olaparib has displayed superior clinical effect in metastatic castration-resistant prostate cancer (mCRPC) patients with the homologous recombination repair (HRR) genes mutations. However, when a patient's tumor tissue volume is insufficient for genomic profiling of HRR gene mutations, circulating tumor DNA (ctDNA) may be useful in helping to determine and monitor the efficacy of olaparib, as well as in abiraterone-combination treatment, and for understanding any resistance mechanism related to such mutations. CASE SUMMARY: A 61-year-old man who was diagnosed with metastatic prostate adenocarcinoma was initially hormone sensitivity, showing high Gleason score (5 + 5 = 10) and absolute positive rate (14/14 biopsied specimens). Following failure of several standard therapies, the patient progressed to mCRPC. Surprisingly, the patient showed good response to olaparib-abiraterone-prednisone combination treatment (an androgen-deprivation therapy, provided as the 'final choice' in China). Serum total prostate-specific antigen (TPSA) level reduced and symptoms remitted for 4 months. However, thereafter, serum TPSA levels began slowly increasing, indicating development of olaparib resistance. Subsequent comprehensive genomic profiling of ctDNA, screening 508 cancer-related genes by next-generation sequencing, identified 10 somatic variants as well as 3 copy number alterations. Two identified reverse missense mutations in partner and localizer of BRCA2 (PALB2) may have recovered the reading frame, restoring function of the primary germline PALB2 mutation and causing resistance to the PARP inhibitor olaparib. CONCLUSION: Reverse mutations in PALB2, discovered via genomic profiling of ctDNA, may represent a potential resistance mechanism against olaparib in mCRPC.
RESUMEN
As all-trans retinoic acid (ATRA) and arsenic trioxide (ATO) are widely accepted in treating acute promyelocytic leukemia (APL), deescalating toxicity becomes a research hotspot. Here, we evaluated whether chemotherapy could be replaced or reduced by ATO in APL patients at different risks. After achieving complete remission with ATRA-ATO-based induction therapy, patients were randomized (1:1) into ATO and non-ATO groups for consolidation: ATRA-ATO versus ATRA-anthracycline for low-/intermediate-risk patients, or ATRA-ATO-anthracycline versus ATRA-anthracycline-cytarabine for high-risk patients. The primary end point was to assess disease-free survival (DFS) at 3 y by a noninferiority margin of -5%; 855 patients were enrolled with a median follow-up of 54.9 mo, and 658 of 755 patients could be evaluated at 3 y. In the ATO group, 96.1% (319/332) achieved 3-y DFS, compared to 92.6% (302/326) in the non-ATO group. The difference was 3.45% (95% CI -0.07 to 6.97), confirming noninferiority (P < 0.001). Using the Kaplan-Meier method, the estimated 7-y DFS was 95.7% (95% CI 93.6 to 97.9) in ATO and 92.6% (95% CI 89.8 to 95.4) in non-ATO groups (P = 0.066). Concerning secondary end points, the 7-y cumulative incidence of relapse (CIR) was significantly lower in ATO (2.2% [95% CI 1.1 to 4.2]) than in non-ATO group (6.1% [95% CI 3.9 to 9.5], P = 0.011). In addition, grade 3 to 4 hematological toxicities were significantly reduced in the ATO group during consolidation. Hence, ATRA-ATO in both chemotherapy-replacing and -reducing settings in consolidation is not inferior to ATRA-chemotherapy (https://www.clinicaltrials.gov/, NCT01987297).
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Trióxido de Arsénico/administración & dosificación , Leucemia Promielocítica Aguda/tratamiento farmacológico , Tretinoina/administración & dosificación , Adulto , Anciano , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Trióxido de Arsénico/efectos adversos , Quimioterapia de Consolidación/efectos adversos , Citarabina/administración & dosificación , Citarabina/efectos adversos , Supervivencia sin Enfermedad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Inducción de Remisión , Resultado del Tratamiento , Tretinoina/efectos adversosRESUMEN
Bortezomib is a novel proteasome inhibitor, which has been successfully used to treat mantle cell lymphoma and multiple myeloma. However, the direct effects of bortezomib on acute promyelocytic leukaemia (APL) have not been fully investigated. In the present study, the WST-8 assay, western blotting, flow cytometry, monodansylcadaverine staining and transmission electron microscopy were performed. It was demonstrated that bortezomib treatment induced a time- and dose-dependent decrease in the viability of NB4 cells. Bortezomib treatment induced cell apoptosis in NB4 cells, as assessed by Annexin V/propidium iodide analysis, and the detection of cleaved caspase-3, cleaved poly(ADP-ribose) polymerase, Bax and Bcl-2 expression. Furthermore, bortezomib treatment induced autophagy in NB4 cells, as indicated by autophagosome formation, p62 degradation, LC3-I to LC3-II conversion and formation of acidic autophagic vacuoles. Notably, autophagy induced by bortezomib was initiated prior to apoptosis. Inhibition of autophagy by knocking down Beclin-1 expression increased bortezomib-induced apoptosis in NB4 cells. Therefore, the present study revealed that the combination of bortezomib and autophagy inhibition may be a potential treatment strategy for APL.
RESUMEN
OBJECTIVE: To investigate the predictive value of neutrophil/lymphocyte ratio (NLR) and platelet/lymphocyte ratio (PLR) for the patients with diffuse large B-cell lymphoma (DLBCL). METHODS: The clinical data of 57 DLBCL patients admitted in the First Affiliated hospital of Anhui Medical University were analyzed retrospectively. According to ROC curve, the cut-off value for NLR and PLR was deterimined, and the patients were divided into high and low NLR/PLR groups before first chamotherapy. Then the relation of NLR and PLR with overall survival (OS) and progression-free survival (PFS) was analyzed by univariate and multivariate COX regression. RESULTS: The optimal cut-off value for NLR and PLR was 2.915 and 270.27, respectively. NLR at the diagnosis was found to be an independent predictor for OS and PFS by univariate and multivariate analysis, while the PLR was an independent predictor for PFS, but did not affect the OS. CONCLUSION: NLR and PLR may provide additional prognostic information for DLBCL patients.
Asunto(s)
Plaquetas/citología , Linfocitos/citología , Linfoma de Células B Grandes Difuso/diagnóstico , Neutrófilos/citología , Supervivencia sin Enfermedad , Humanos , Recuento de Linfocitos , Análisis Multivariante , Pronóstico , Estudios RetrospectivosRESUMEN
A simple and highly-efficient approach to monitor the expression of P-glycoprotein (P-gp) in cells was urgently needed to demonstrate the drug resistance of cancer cells. Herein, a competitive method-based electrochemiluminescent (ECL) assay with a single ECL indicator was proposed for the first time to efficiently estimate the concentration ratio of two proteins. By converting the different proteins to partially coincident nucleotide sequences via a sandwich type immunoassay on magnetic beads, the concentration ratio related ECL signals could be obtained via competitive nucleotide hybridization on an electrode surface. This method could thoroughly overcome the limitations of simultaneous ECL assays via multiple ECL indicators with inevitable cross reactions. At the same time, rolling circle amplification was employed to improve the detection performances, especially the detection limit and sensitivity. With P-gp and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as a model, the proposed ECL assay was successfully employed to monitor the drug resistance of cancer cells. Compared with conventional technologies, improved sensitivity and accuracy were achieved with a correlation coefficient of 0.9928 and a detection limit of 0.52%. Success in the establishment of the competitive method-based ECL assay offered an efficient strategy to demonstrate the concentration ratio of two proteins and a potential approach for detecting other proteins and nucleotide sequences, revealing a new avenue for ultrasensitive biomolecule diagnostics, especially in cell function research.
RESUMEN
BACKGROUND: MicroRNAs (miR, miRNAs) play pivotal roles in numerous physiological and pathophysiological contexts. We investigated whether miR-362-5p act as an oncogene in chronic myeloid leukaemia (CML) and aimed to understand its potential underlying mechanisms. METHODS: We compared the miR-362-5p expression levels between CML and non-CML cell lines, and between fresh blood samples from CML patients and normal healthy controls using quantitative real-time PCR (qPCR). Cell counting kit-8 (CCK-8) and Annexin V-FITC/PI analyses were used to measure the effects of miR-362-5p on proliferation and apoptosis, and Transwell assays were used to evaluate migration and invasion. A xenograft model was used to examine in vivo tumourigenicity. The potential target of miR-362-5p was confirmed by a luciferase reporter assay, qPCR and western blotting. Involvement of the JNK1/2 and P38 pathways was investigated by western blotting. RESULTS: miR-362-5p was up-regulated in CML cell lines and fresh blood samples from CML patients, and was associated with Growth arrest and DNA damage-inducible (GADD)45α down-regulation. Inhibition of miR-362-5p simultaneously repressed tumour growth and up-regulated GADD45α expression in a xenograft model. Consistently, the knockdown of GADD45α expression partially neutralized the effects of miR-362-5p inhibition. Furthermore study suggested that GADD45α mediated downstream the effects of miR-362-5p, which might indirectly regulates the activation of the JNK1/2 and P38 signalling pathways. CONCLUSION: miR-362-5p acts as an oncomiR that down-regulates GADD45α, which consequently activates the JNK1/2 and P38 signalling. This finding provides novel insights into CML leukaemogenesis and may help identify new diagnostic and therapeutic targets.
Asunto(s)
Proteínas de Ciclo Celular/metabolismo , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , MicroARNs/fisiología , Proteínas Nucleares/metabolismo , Animales , Apoptosis/genética , Apoptosis/fisiología , Ciclo Celular/genética , Ciclo Celular/fisiología , Proteínas de Ciclo Celular/genética , Línea Celular Tumoral , Movimiento Celular/genética , Movimiento Celular/fisiología , Proliferación Celular/genética , Proliferación Celular/fisiología , Regulación hacia Abajo , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Regulación Neoplásica de la Expresión Génica/fisiología , Células HEK293 , Humanos , Células K562 , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , MicroARNs/genética , Proteínas Nucleares/genética , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
OBJECTIVE: To detect the expression of DNA methyltransferases (DNMT) mRNA in the patients with acute myelogenous leukemia (AML) and to analyze the retationship between the mRNA expression of DNMT and cellular and moleculogenetic risk stratifieation in AML patients, and to evaluate the role of the DNMT mRNA expression in AML prognosis and clinical treatment. METHODS: The mRNA expression of DNMT was detected by real-time PCR in 123 AML patients and 20 healthy people. RESULTS: the mRNA expression levels of DNMT were lower in the healthy people and higher in AML patients; the mRNA expression levels of DNMT in the patients after the consolidation therapy were lower than that in the patients of initial diagnosis and replapse; The mRNA expression levels of DNMT did not correlate with age, sex and the clinical characteristics at initial diagnosis, such as white blood cell count, FAB classification and chromosomal karyotype in AML patients. In CR patients after standard treatment, the initial mRNA expression level of DNMT3b was higher. Based on cellular and moleculogenetic risk stratificantion, the DNMT expression level in the intermediate risk AML patients was higher. CONCLUSION: The mRNA expression of DNMT may play an important role in AML pathogenesis and can serve as an index for evaluating AML prognosis and for instructing clinical treatment.
Asunto(s)
Regulación Leucémica de la Expresión Génica , Leucemia Mieloide Aguda , ADN , ADN (Citosina-5-)-Metiltransferasas , Metilación de ADN , Humanos , Cariotipificación , Pronóstico , ARN Mensajero , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Land-use change has a crucial influence on soil respiration, which further affects soil nutrient availability and carbon stock. We monitored soil respiration rates under different land-use types (tea gardens with three production levels, adjacent woodland, and a vegetable field) in Eastern China at weekly intervals over a year using the dynamic closed chamber method. The relationship between soil respiration and environmental factors was also evaluated. The soil respiration rate exhibited a remarkable single peak that was highest in July/August and lowest in January. The annual cumulative respiration flux increased by 25.6% and 20.9% in the tea garden with high production (HP) and the vegetable field (VF), respectively, relative to woodland (WL). However, no significant differences were observed between tea gardens with medium production (MP), low production (LP), WL, and VF. Soil respiration rates were significantly and positively correlated with organic carbon, total nitrogen, and available phosphorous content. Each site displayed a significant exponential relationship between soil respiration and soil temperature measured at 5 cm depth, which explained 84-98% of the variation in soil respiration. The model with a combination of soil temperature and moisture was better at predicting the temporal variation of soil respiration rate than the single temperature model for all sites. Q10 was 2.40, 2.00, and 1.86-1.98 for VF, WL, and tea gardens, respectively, indicating that converting WL to VF increased and converting to tea gardens decreased the sensitivity of soil respiration to temperature. The equation of the multiple linear regression showed that identical factors, including soil organic carbon (SOC), soil water content (SWC), pH, and water soluble aluminum (WSAl), drove the changes in soil respiration and Q10 after conversion of land use. Temporal variations of soil respiration were mainly controlled by soil temperature, whereas spatial variations were influenced by SOC, SWC, pH, and WSAl.
Asunto(s)
Productos Agrícolas/clasificación , Productos Agrícolas/crecimiento & desarrollo , Ecosistema , Suelo/química , China , Estaciones del Año , TemperaturaRESUMEN
IFN regulatory factor 4 binding protein (IBP) has been shown to play an important role in the progression of malignant tumors such as breast cancer cells, but its function in oral squamous cell carcinoma (OSCC) remains unclear. We found that IBP ectopically expressed in some OSCC specimens but not in normal oral mucosa epithelium tissues. IBP expression was significantly correlated with tumor size, differentiation, clinical stage, and distant metastasis. Furthermore, IBP markedly promoted OSCC cell proliferation, shortened the G1 interval in the cell cycle, and increased cyclin D1 expression. These findings suggest that IBP may be a potential therapeutic target for OSCC.
Asunto(s)
Biomarcadores de Tumor/agonistas , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patología , Proteínas de Unión al ADN/biosíntesis , Factores de Intercambio de Guanina Nucleótido/biosíntesis , Neoplasias de la Boca/metabolismo , Neoplasias de la Boca/patología , Proteínas Nucleares/biosíntesis , Animales , Western Blotting , Línea Celular Tumoral , Proliferación Celular , Femenino , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Humanos , Inmunohistoquímica , Masculino , Ratones , Ratones Endogámicos BALB C , Estadificación de Neoplasias , Análisis de Matrices Tisulares , Transfección , Trasplante HeterólogoRESUMEN
OBJECTIVE: To evaluate the efficacy and safety of thrombopoietin (TPO) on platelet engraftment in hematological malignancies patients after allogeneic haematopoietic stem cell transplantation (allo-HSCT). METHODS: One hundred and twenty patients were enrolled in a multicenter, open-label, randomized, controlled clinical trial, and were randomized into 4 treatment groups following allo-HSCT. Group A was the control arm without TPO, while group B, C and D were trial arms with received 300 U×kg(-1)×d(-1) of TPO starting from day +1, +4 and +7, respectively. A total of 89 cases were evaluated, of which 22 cases in group A, 23 in group B, 20 in group C and 24 in group D. Efficacy evaluation (the time of platelet engraftment, the number of platelet transfusion) and safety evaluation \[adverse events, routine blood tests, liver and renal function, coagulation function and occurrence of graft-versus-host disease (GVHD)\] were observed. RESULTS: The median platelet engraftment time in experimental groups (groups B, C and D) were on day (13.17 ± 2.89), day (12.15 ± 2.08), day (12.33 ± 1.76), respectively, and that in control group was on day (14.82 ± 5.05). There was statistically significant difference between two groups (P = 0.029), There were no statistically significant difference in the average amount of platelet transfusion, platelet engraftment time, and platelet nadir value among the 3 experimental groups. No significant adverse events were observed in experimental groups. CONCLUSIONS: TPO administration following allo-HSCT for patients with hematologic malignancies appears to shorten platelet engraftment time. TPO given starting from day +7 is effective and safe.
Asunto(s)
Neoplasias Hematológicas/cirugía , Trasplante de Células Madre Hematopoyéticas , Transfusión de Plaquetas/métodos , Trombopoyetina/uso terapéutico , Adolescente , Adulto , Plaquetas , Niño , Femenino , Humanos , Masculino , Persona de Mediana Edad , Trasplante Homólogo , Adulto JovenRESUMEN
OBJECTIVE: To investigate the effect of total astragalosides (TA) on proliferation and apoptosis in human leukemia NB4 cells in vitro. METHODS: The NB4 cells were treated with TA at different concentrations for 48 h in culture. Growth inhibition rates were measured by CCK-8 method. Flow cytometry was used to explore the cell apoptosis and the activity of NF-κB and Akt during apoptosis. RESULTS: TA at different concentrations (200, 400, 600, 800 mg/L) inhibited proliferation of NB4 cells in a dose-dependent manner (P < 0.05), and the inhibitory rates of TA on NB4 cells were (14.54 ± 3.20)%, (24.79 ± 3.98)%, (57.28 ± 4.71)% and (88.28 ± 4.65)%, respectively. In terms of the induction of apoptosis, there was a significant difference between the TA group and blank control [(1.80 ± 1.24)%, P < 0.05]. At TA doses of 200, 400 and 600 mg/L, the apoptotic rates of NB4 cells were (10.03 ± 3.31)%, (14.87 ± 3.65)%, (23.45 ± 1.90)%, respectively. Besides, TA induced apoptosis of NB4 cells in a dose-dependent manner in the groups of 200 mg/L, 400 mg/L, 600 mg/L (P < 0.05). But there was no significant difference in apoptotic rates between the groups of 800 mg/L and 600 mg/L [(23.45 ± 1.90)%, P > 0.05]. In the group of 800 mg/L, the necrotic cells increased highly and the necrotic rate reached (45.65 ± 3.16)%. After TA treatment of NB4 cells at different concentrations (200, 400, 600 mg/L), the expression of NF-κB protein was significantly decreased compared with that of the blank control (9.79 ± 0.95, P < 0.05), while Akt protein was not significantly decreased (P > 0.05). CONCLUSION: TA can inhibit the growth of NB4 cells and induce apoptosis in NB4 cells through an Akt-independent NF-κB signaling pathway.
Asunto(s)
Apoptosis/efectos de los fármacos , Astragalus propinquus/química , Medicamentos Herbarios Chinos/farmacología , Leucemia Promielocítica Aguda/patología , Saponinas/farmacología , Antineoplásicos Fitogénicos/administración & dosificación , Antineoplásicos Fitogénicos/aislamiento & purificación , Antineoplásicos Fitogénicos/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/aislamiento & purificación , Humanos , Leucemia Promielocítica Aguda/metabolismo , FN-kappa B/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Saponinas/administración & dosificación , Saponinas/aislamiento & purificaciónRESUMEN
Dehydrins (DHNs), proteins with protective functions encoded by the late embryogenesis abundant (LEA), are differentially up-regulated at the transcriptional level under environmental stresses such as water deficit, salinity, and low temperature or in response to ABA. Data involving protein structure and expression of dehydrins could elucidate their functional roles under dehydration conditions in plants. Dhn6 gene of 1767 bp with an intron (92 bp) in six-rowed barley, a member of LEA D11, was cloned into pMD18-T vector in the present study. It shared 93.18% identity with Hordeum vulgare ssp. vulgare (GenBank accession: AF043091), encoding a protein composed of 523 amino acid residues with the predicted molecular weight of 49.68 kDa and the theoretical isoelectric point of 8.04. Analyses of domain and structure indicated that the protein was dominantly composed of 83% hydrophillic amino acid residues, with numerous imperative curls and three loosed helixes. Three-dimensional measurement revealed that the water-soluble lipid-associating protein was attributed to its twisted cable formed by reverse paralleled chains. Moreover, the putative amphipathic α-helices formed by K-segments might play roles in protecting membrane structure in barley. Significantly, relatively high accumulations of Dhn6 gene were detected after 8 h of water deficit by real-time quantitative RT-PCR. These results possibly suggested that the accumulation of Dhn6 gene could be critical for resistant dehydration in plants.
Asunto(s)
Sequías , Regulación de la Expresión Génica de las Plantas , Hordeum/genética , Proteínas de Plantas/química , Proteínas de Plantas/genética , Secuencia de Aminoácidos , Cruzamiento , Clonación Molecular , Biología Computacional , Hordeum/fisiología , Modelos Moleculares , Datos de Secuencia Molecular , Proteínas de Plantas/metabolismo , Estructura Secundaria de Proteína , Análisis de Secuencia de ADN , Estrés Fisiológico/genéticaRESUMEN
This study was purposed to evaluate ABL tyrosine kinase point mutations in imatinib-treated chronic myeloid leukemia (CML) patients and their clinical significance. 51 bone marrow samples from 28 imatinib-resistant patients and 10 newly diagnosed CML patients were collected. ABL kinase domain of bcr-abl allele was amplified by nested reverse transcription-polymerase chain reaction, followed by purifying, directly sequencing and sequence homology analysis of amplified products in order to determine the existence and type of point mutation. The results showed that the point mutations were found in 12 of 38 patients, and all the 12 ones progressed to advanced disease or death. 2 patients showed Met351Thr mutation, 7 patients showed Glu252His, 2 patients showed Glu279Lys, the other types were Glu255Val and Glu355Gly, each of which was tested in one patient. The incidence of the point mutation was 17.6%, 45.5% and 44.4% in chronic, accelerated and blast phase respectively. The incidences of point mutation in hematologically and genetically resistant patients were 50% (5/10) and 44.4% (8/18), and the 95% confidence interval (CI) was 12.3% - 87.7% and 19% - 69.9% respectively. It is concluded that ABL kinase point mutation is an important mechanism of imatinib resistance, monitoring the ABL kinase domain point mutation is helpful to estimating the prognosis and adjusting the therapeutic strategy.
Asunto(s)
Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Mutación Puntual , Proteínas Tirosina Quinasas/genética , Adulto , Anciano , Benzamidas , Femenino , Humanos , Mesilato de Imatinib , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Masculino , Persona de Mediana Edad , Piperazinas/uso terapéutico , Pirimidinas/uso terapéutico , Resultado del Tratamiento , Adulto JovenRESUMEN
OBJECTIVE: To explore the impact of IL-2- and IL-15-activated donor natural killer (NK) cell infusion on graft-versus-host-disease (GVHD) and graft-versus-leukemia (GVL) effect post allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: The C57BL/6 mice splenic NK cells were selected by microbeads, and then expanded in the media containing IL-2 and IL-15. The killing activity of NK cells was detected. In the leukemia mouse model, recipients (BALB/c) were intravenously inoculated with EL9611 leukemia cells 8 days before transplantation. Lethally irradiated BALB/c recipient mice were transplanted with 5 x 10(6) bone marrow cells (BMCs), or 5 x 10(6) BMCs plus 1 x 10(7) splenocytes with or without 1 x 10(7) activated NK cells. Additionally, NK cell infusion group mice were intraperitoneally injected with a mixture of IL-2 and IL-15 post transplant. Survival time, GVHD occurrence, lineage chimerism, TRBV spectra-typing were observed post transplant. RESULTS: The purity of isolated splenic NK cells was 95.7% - 97.1%. The killing activity of NK cells after activation was increased by 3 times. GVHD did not occurred in allogeneic BMCs infusion group, whereas did from 1 week after transplant in allogeneic BMCs + splenocytes infusion group. The severity of GVHD in total body irradiation (TBI) experimental group was significantly lower than in splenocytes infusion group (P < 0.05). The survival time was 9.5 - 14.0 d in TBI alone conditioning group. In leukemia mouse model, 100 day survival rate was 10% the rest of them were died of leukemia while in experimental group, the more than 100 days survival rate was 80% (P < 0.01). PB NK cells at 2 week post-transplant were 4.8% in experimental group and 2.8% in control group. NK cells recovery in experimental group was earlier than that in control group (P < 0.05). TRBV reconstitution was faster in experimental group than in control group, moreover, the number of TRBV family expression was more in experimental group than in control group which mainly expressed monoclone or oligo-clone. CONCLUSIONS: Donor alloreactive NK cells can be efficiently expanded and activated with IL-2 and IL-15. Donor activated NK cell infusion and IL-2, IL-15 treatment can promote immune reconstitution, mitigate GVHD and reduce leukemia relapse.
Asunto(s)
Trasplante de Células Madre Hematopoyéticas , Interleucina-15/inmunología , Interleucina-2/inmunología , Células Asesinas Naturales/inmunología , Animales , Células Cultivadas , Enfermedad Injerto contra Huésped/prevención & control , Efecto Injerto vs Leucemia , Interleucina-15/farmacología , Interleucina-2/farmacología , Células Asesinas Naturales/citología , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BLRESUMEN
OBJECTIVE: To investigate the effect of HLA-Cw on haploidentical hematopoietic stem cell transplantation (HHSCT) without T-cell depletion. METHODS: HLA-Cw were detected with PCR-SSP, the clinical data of 21 cases of haploidentical hematopoietic stem cell transplantation, including 8 standard risk and 13 high risk cases from July 2002 to March 2006 were summarized, and the effect of HLA-Cw in HHSCT was analyzed. RESULTS: Twenty patients achieved sustained, full-donor-type engraftment. The HLA-Cw matched and mismatched groups attained neutrophil recovery at a median of 12 days and 13 days, and platelet recovery to more than 20 x 10(9)/L at a median of 20 days and 23 days respectively (P > 0.05). The cumulative incidences of grades II-IV acute GVHD were 76.9% in HLA-Cw matched group and 14.3% in the mismatched group(P < 0.05). The incidences of chronic GVHD were 85.7% in HLA-Cw matched group and 57.1% in the mismatched group(P > 0.05). The 28 months disease-free survival probabilities were 49.0% in HLA-Cw matched group, and 85.7% in the mismatched group (P > 0.05). The Karnofsky score of survival patients was over 90%. CONCLUSION: HLA-Cw mismatched in donor and recipient of HHSCT is beneficial for reducing II-IV aGVHD, and being in favor of long term survival.
Asunto(s)
Antígenos HLA-C/inmunología , Trasplante de Células Madre Hematopoyéticas , Adolescente , Adulto , Niño , Femenino , Estudios de Seguimiento , Enfermedad Injerto contra Huésped/inmunología , Humanos , Masculino , Persona de Mediana Edad , Tasa de Supervivencia , Trasplante Homólogo/inmunologíaRESUMEN
OBJECTIVE: To investigate the effects and prognosis of malignant hematological disease after HLA haploidentical hematopoietic stem cell transplantation (H-HSCT) without T-cell depletion. METHODS: The clinical data of 31 cases with malignant hemopoietic disease treated with H-HSCT from July 2002 to July 2006 were analyzed, including 11 cases of standard risk and 20 of high risk. RESULTS: 30 patients achieved engraftment of a median of 13 and 22 days for neutrophil and platelet, with an accumulative incidence of II - IV grade acute graft-versus-host disease (GVHD) 61.3%, and an accumulative incidence of chronic GVHD 41.9%. 13 patients survived with Karnofsky scale over 90.0% after a median follow-up of 24 months. 33 months of accumulative survival was 62.3% in the standard risk group and 35.0% in the high risk group. The CD(3)(+) T cells count of the graft and the disparity of HLA-A, B, DR loci were the major factors of impact on acute GVHD. CONCLUSION: HLA H-HSCT is an effective therapeutic method for malignant hematological disease, CD(3)(+) T cells count of the graft and the disparity of HLA-A, B, DR loci are the major factors of impact on acute GVHD.