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BACKGROUND: The endothelial-to-hematopoietic transition (EHT) process during definitive hematopoiesis is highly conserved in vertebrates. Stage-specific expression of transposable elements (TEs) has been detected during zebrafish EHT and may promote hematopoietic stem cell (HSC) formation by activating inflammatory signaling. However, little is known about how TEs contribute to the EHT process in human and mouse. RESULTS: We reconstructed the single-cell EHT trajectories of human and mouse and resolved the dynamic expression patterns of TEs during EHT. Most TEs presented a transient co-upregulation pattern along the conserved EHT trajectories, coinciding with the temporal relaxation of epigenetic silencing systems. TE products can be sensed by multiple pattern recognition receptors, triggering inflammatory signaling to facilitate HSC emergence. Interestingly, we observed that hypoxia-related signals were enriched in cells with higher TE expression. Furthermore, we constructed the hematopoietic cis-regulatory network of accessible TEs and identified potential TE-derived enhancers that may boost the expression of specific EHT marker genes. CONCLUSIONS: Our study provides a systematic vision of how TEs are dynamically controlled to promote the hematopoietic fate decisions through transcriptional and cis-regulatory networks, and pre-train the immunity of nascent HSCs.
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Elementos Transponibles de ADN , Hematopoyesis , Células Madre Hematopoyéticas , Análisis de la Célula Individual , Animales , Elementos Transponibles de ADN/genética , Análisis de la Célula Individual/métodos , Ratones , Hematopoyesis/genética , Humanos , Células Madre Hematopoyéticas/metabolismo , Células Endoteliales/metabolismoRESUMEN
INTRODUCTION: This systematic review and meta-analysis aims to explore the influence of ferumoxytol versus placebo on iron deficiency anemia. METHODS: We search for randomized controlled trials (RCTs) assessing the effect of ferumoxytol on iron deficiency anemia on PubMed, EMbase, Web of science, EBSCO, and Cochrane library databases. This meta-analysis is performed using the random-effects model. RESULTS: Four RCTs are included in the meta-analysis. Compared with the control group for iron deficiency anemia, intravenous ferumoxytol can significantly improve the proportion of patients with a ≥20 g/L hemoglobin (Hb) increase (RR = 18.43; 95% CI = 7.29-46.57; p < 0.00001), the proportion of patients with an Hb level ≥120 g/L (RR = 18.55; 95% CI = 8.66-39.72; p < 0.00001), transferrin saturation (mean difference = 11.08; 95% CI = 9.86-12.31; p < 0.00001) and FACIT-fatigue score (mean difference = 4.60; 95% CI = 3.21-6.00; p < 0.00001), but has no remarkable influence on adverse events (RR = 1.33; 95% CI = 0.84-2.10; p = 0.22), serious adverse events (RR = 1.22; 95% CI = 0.74-2.02; p = 0.44), and death (RR = 0.32; 95% CI = 0.05-1.95; p = 0.22). CONCLUSIONS: Intravenous ferumoxytol can provide the important benefits for iron deficiency anemia.
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Anemia Ferropénica , Óxido Ferrosoférrico/uso terapéutico , Anemia Ferropénica/sangre , Anemia Ferropénica/tratamiento farmacológico , Anemia Ferropénica/fisiopatología , Óxido Ferrosoférrico/efectos adversos , Hemoglobinas/metabolismo , Humanos , Ensayos Clínicos Controlados Aleatorios como AsuntoRESUMEN
BACKGROUND T cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematologic malignancy caused by abnormal proliferation of immature T cell progenitors. Chemotherapy of T-ALL usually consists of induction, consolidation, and long-term maintenance. Diacetyl hexamethylene diamine (CAHB) is a newly developed agent that induces the differentiation of malignant cells and deprives their clonal growth ability. Since its effect on T-ALL has not been previously determined, we evaluated its potential function in the Jurkat cell line. MATERIAL AND METHODS MTT assay was conducted to evaluate the cytotoxicity and anti-proliferative effect of CAHB. The apoptosis level of CAHB-treated Jurkat cells was evaluated using flow cytometry via staining with Annexin V/PI or cleaved-caspase-3. The alteration of mitochondrial membrane potential was determined by flow cytometry. The expression of Bax and Bcl-2 was evaluated by RT-PCR and Western blot. Western blot was also used to assess the activation of Akt. RESULTS CAHB inhibited the proliferation and promoted the apoptosis of Jurkat cells in a time- and dose-dependent manner by decreasing activation of Akt, reducing the mitochondrial membrane potential, and downregulating the Bcl-2/Bax ratio. CONCLUSIONS Our data suggest that CAHB might be regarded as a novel treatment agent for T-ALL since it can induce apoptosis and inhibit proliferation of the T-ALL cell line at a relatively low level.
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Diaminas/farmacología , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo , Antineoplásicos Fitogénicos/farmacología , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Diacetil , Diaminas/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Humanos , Células Jurkat , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Fosfatidilinositol 3-Quinasas/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamiento farmacológico , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
The effect of Sargassum pallidum (brown seaweed) aqueous extract on the immunity function and antioxidant activities in was studied gastric cancer rats. Treatment with Sargassum pallidum aqueous extract at oral doses 400, 600 or 800 mg/kg body weight was found to provide a dose-dependent protection against N-methyl-N'-nitro-Nnitrosoguanidine (MNNG)-induced immunity damage and oxidative injury by enhancing serum interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-10 (IL-10) levels, decreasing interleukin-6 (IL-6), interleukin-1ß (IL-1ß), tumor necrosis factor-alpha (TNF-α) levels, preserving normal antioxidant enzymes activities, and by inhibiting lipid peroxidation in gastric mucosa. It can be concluded that Sargassum pallidum aqueous extract may enhance the immunity and antioxidant activities in gastric cancer rats.
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Antioxidantes/uso terapéutico , Inmunidad , Extractos Vegetales/uso terapéutico , Sargassum/química , Neoplasias Gástricas/tratamiento farmacológico , Neoplasias Gástricas/inmunología , Agua/química , Animales , Antioxidantes/farmacología , Catalasa/metabolismo , Mucosa Gástrica/efectos de los fármacos , Mucosa Gástrica/enzimología , Mucosa Gástrica/patología , Glutatión/sangre , Glutatión Peroxidasa/metabolismo , Inmunidad/efectos de los fármacos , Interleucinas/sangre , Masculino , Malondialdehído/metabolismo , Fitoterapia , Extractos Vegetales/farmacología , Ratas , Ratas Wistar , Neoplasias Gástricas/sangre , Superóxido Dismutasa/metabolismoRESUMEN
Background: Diffuse large B-cell lymphoma (DLBCL), which is considered to be the most common subtype of lymphoma, is an aggressive tumor. Necroptosis, a novel type of programmed cell death, plays a bidirectional role in tumors and participates in the tumor microenvironment to influence tumor development. Targeting necroptosis is an intriguing direction, whereas its role in DLBCL needs to be further discussed. Methods: We obtained 17 DLBCL-associated necroptosis-related genes by univariate cox regression screening. We clustered in GSE31312 depending on their expressions of these 17 genes and analyzed the differences in clinical characteristics between different clusters. To investigate the differences in prognosis across distinct clusters, the Kaplan-Meier method was utilized. The variations in the tumor immune microenvironment (TME) between distinct necroptosis-related clusters were investigated via "ESTIMATE", "Cibersort" and single-sample geneset enrichment analysis (ssGSEA). Finally, we constructed a 6-gene prognostic model by lasso-cox regression and subsequently integrated clinical features to construct a prognostic nomogram. Results: Our analysis indicated stable but distinct mechanism of action of necroptosis in DLBCL. Based on necroptosis-related genes and cluster-associated genes, we identified three groups of patients with significant differences in prognosis, TME, and chemotherapy drug sensitivity. Analysis of immune infiltration in the TME showed that cluster 1, which displayed the best prognosis, was significantly infiltrated by natural killer T cells, dendritic cells, CD8+ T cells, and M1 macrophages. Cluster 3 presented M2 macrophage infiltration and the worst prognosis. Importantly, the prognostic model successfully differentiated high-risk from low-risk patients, and could forecast the survival of DLBCL patients. And the constructed nomogram demonstrated a remarkable capacity to forecast the survival time of DLBCL patients after incorporating predictive clinical characteristics. Conclusion: The different patterns of necroptosis explain its role in regulating the immune microenvironment of DLBCL and the response to R-CHOP treatment. Systematic assessment of necroptosis patterns in patients with DLBCL will help us understand the characteristics of tumor microenvironment cell infiltration and aid in the development of tailored therapy regimens.
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BACKGROUND: Multiple myeloma (MM) is a malignant hematopoietic disease that is usually incurable. RNA-binding proteins (RBPs) are involved in the development of many tumors, but their prognostic significance has not been systematically described in MM. Here, we developed a prognostic signature based on eight RBP-related genes to distinguish MM cohorts with different prognoses. METHOD: After screening the differentially expressed RBPs, univariate Cox regression was performed to evaluate the prognostic relevance of each gene using The Cancer Genome Atlas (TCGA)-Multiple Myeloma Research Foundation (MMRF) dataset. Lasso and stepwise Cox regressions were used to establish a risk prediction model through the training set, and they were validated in three Gene Expression Omnibus (GEO) datasets. We developed a signature based on eight RBP-related genes, which could classify MM patients into high- and low-score groups. The predictive ability was evaluated using bioinformatics methods. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and gene set enrichment analyses were performed to identify potentially significant biological processes (BPs) in MM. RESULT: The prognostic signature performed well in the TCGA-MMRF dataset. The signature includes eight hub genes: HNRNPC, RPLP2, SNRPB, EXOSC8, RARS2, MRPS31, ZC3H6, and DROSHA. Kaplan-Meier survival curves showed that the prognosis of the risk status showed significant differences. A nomogram was constructed with age; B2M, LDH, and ALB levels; and risk status as prognostic parameters. Receiver operating characteristic (ROC) curve, C-index, calibration analysis, and decision curve analysis (DCA) showed that the risk module and nomogram performed well in 1, 3, 5, and 7-year overall survival (OS). Functional analysis suggested that the spliceosome pathway may be a major pathway by which RBPs are involved in myeloma development. Moreover, our signature can improve on the R-International Staging System (ISS)/ISS scoring system (especially for stage II), which may have guiding significance for the future. CONCLUSION: We constructed and verified the 8-RBP signature, which can effectively predict the prognosis of myeloma patients, and suggested that RBPs are promising biomarkers for MM.
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Background: Diffuse large B-cell lymphoma (DLBCL) is a common aggressive B-cell non-Hodgkin lymphoma (B-NHL). While combined chemotherapy has improved the outcomes of DLBCL, it remains a highly detrimental disease. Pyroptosis, an inflammatory programmed cell death, is considered to have both tumor-promoting and tumor-suppressing effects. The role of pyroptosis in DLBCL has been gradually appreciated, but its value needs further investigation. Methods: We analyzed mutations and copy number variation (CNV) alterations of pyroptosis-related genes (PRGs) from The Cancer Genome Atlas (TCGA) cohort and evaluated the differences in expression in normal B cells and DLBCL patients in two Gene Expression Omnibus (GEO) datasets (GSE12195 and GSE56315). Based on the expression of 52 PRGs, we divided 421 DLBCL patients from the GSE31312 dataset into distinct clusters using consensus clustering. The Kaplan-Meier method was used to prognosis among the three clusters, and GSVA was used to explore differences in the biological functions. ESTIMATE and single-sample gene-set enrichment analysis (ssGSEA) were used to analyze the tumor immune microenvironment (TME) in different clusters. A risk score signature was developed using a univariate survival analysis and multivariate regression analysis, and the reliability and validity of the signature were verified. By combining the signature with clinical factors, a nomogram was established to predict the prognosis of DLBCL patients. The alluvial diagram and correlation matrix were used to explore the relationship between pyroptosis risk score, clinical features and TME. Results: A large proportion of PRGs are dysregulated in DLBCL and associated with the prognosis. We found three distinct pyroptosis-related clusters (cluster A, B, and C) that differed significantly with regard to the prognosis, biological process, clinical characteristics, chemotherapeutic drug sensitivity, and TME. Furthermore, we developed a risk score signature that effectively differentiates high and low-risk patients. The nomogram combining this signature with several clinical indicators showed an excellent ability to predict the prognosis of DCBCL patients. Conclusions: This work demonstrates that pyroptosis plays an important role in the diversity and complexity of the TME in DLBCL. The risk signature of pyroptosis is a promising predictive tool. A correct and comprehensive assessment of the mode of action of pyroptosis in individuals will help guide more effective treatment.
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Most randomized trials for acute promyelocytic leukemia (APL) have investigated highly selected patients under idealized conditions, and the findings need to be validated in the real world. We conducted a population-based study of all APL patients in Zhejiang Province, China, with a total population of 82 million people, to assess the generalization of all-trans retinoic acid (ATRA) and arsenic as front-line treatment. The outcomes of APL patients were also analyzed. Between January 2015 and December 2019, 1,233 eligible patients were included in the final analysis. The rate of ATRA and arsenic as front-line treatment increased steadily from 66.2% in 2015 to 83.3% in 2019, with no difference among the size of the center (≥5 or <5 patients per year, p = 0.12) or age (≥60 or <60 years, p = 0.35). The early death (ED) rate, defined as death within 30 days after diagnosis, was 8.2%, and the 3-year overall survival (OS) was 87.9% in the whole patient population. Age (≥60 years) and white blood cell count (>10 × 109/L) were independent risk factors for ED and OS in the multivariate analysis. This population-based study showed that ATRA and arsenic as front-line treatment are widely used under real-world conditions and yield a low ED rate and a high survival rate, which mimic the results from clinical trials, thereby supporting the wider application of APL guidelines in the future.
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Acute myelogenous leukemia (AML) is a class of malignant tumors derived from hematopoietic stem or progenitor cells. The H2.0-like homeobox gene (HLX) encodes transcription factors that function in promoting normal hematopoietic cell proliferation and tumor immunity. The present study analyzed the effect of downregulating the HLX on cell cycle distribution and cell proliferation in AML. Moreover, the current study detected changes in the expression of genes and proteins in the Janus kinase (JAK)/STAT signaling pathway to investigate the mechanism of the action of HLX in tumor immunity in AML. HLX expression in AML cell lines was silenced using small interfering siRNA, and MTS/PMS-assay colorimetric assays were used to assess the effect of knockdown of HLX on AML cell proliferation. Flow cytometry was used to analyze changes in cell cycle distribution, while reverse transcription-quantitative PCR and western blotting were used to detect changes in the expression levels of key components of the JAK/STAT signaling pathway, such as p21-activated kinase 1 (PAK1), neuropilin 1 (NRP1), B-cell translocation gene 1 (BTG1) and STAT5. It was found that HLX was differentially expressed in AML cell lines of various subtypes, and HLX expression was higher in the AML/M3 subtype NB4 cell line compared with the control group. Knockdown of HLX in NB4 cells significantly inhibited cell proliferation and arrested cells in the G0/G1 phase. Moreover, STAT5 protein expression, as well as NRP1 and PAK1 expression levels were downregulated, while BTG1 expression was upregulated when HLX was knocked out by siRNA. Collectively, the results suggested that downregulation of HLX may cause G0/G1 phase arrest and inhibit the proliferation of AML cells by activating the JAK/STAT signaling pathway.
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Objective: To investigate the hyaluronic acid (HA) modified, doxorubicin (DOX) and gallic acid (GA) co-delivered lipid-polymeric hybrid nano-system for leukemia therapy. Methods: We produced a kind of lipid-polymer hybrid nanoparticle (LPHN) with a core-shell structure in which DOX and GA were co-loaded. In vitro and in vivo leukemia therapeutic effects of the HA modified, DOX and GA co-delivered LPHNs (HA-DOX/GA-LPHNs) were evaluated in DOX resistant human HL-60 promyelocytic leukemia cells (HL-60/ADR cells), DOX resistant human K562 chronic myeloid leukemia cells (K562/ADR cells), and HL-60/ADR cells bearing mouse model. Results: The sizes and zeta potentials of HA modified LPHNs were about 160 nm and -40 mV. HA-DOX/GA-LPHNs showed the most prominent cytotoxicity and the best synergistic effect was obtained when DOX/GA ratio was 2/1. In vivo studies revealed that HA-DOX/GA-LPHNs inhibited tumor growth from 956 mm3 to 213 mm3, with an inhibition rate of 77.7%. Conclusion: In summary, the study showed that HA-DOX/GA-LPHNs can be applied as a promising leukemia therapy system.
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Antibióticos Antineoplásicos/farmacología , Doxorrubicina/farmacología , Sistemas de Liberación de Medicamentos , Ácido Gálico/farmacología , Ácido Hialurónico/farmacología , Leucemia Mieloide Aguda/tratamiento farmacológico , Animales , Antibióticos Antineoplásicos/administración & dosificación , Antibióticos Antineoplásicos/química , Proliferación Celular/efectos de los fármacos , Doxorrubicina/administración & dosificación , Doxorrubicina/química , Liberación de Fármacos , Resistencia a Antineoplásicos/efectos de los fármacos , Ensayos de Selección de Medicamentos Antitumorales , Ácido Gálico/administración & dosificación , Ácido Gálico/química , Células HL-60 , Humanos , Ácido Hialurónico/administración & dosificación , Ácido Hialurónico/química , Inyecciones Intravenosas , Células K562 , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/patología , Lípidos/química , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Nanopartículas/química , Neoplasias Experimentales/tratamiento farmacológico , Neoplasias Experimentales/metabolismo , Neoplasias Experimentales/patología , Tamaño de la Partícula , Polímeros/química , Propiedades de SuperficieRESUMEN
BACKGROUND: We studied the expression of CD200 in a series of 101 patients with diagnosis of myelodysplastic syndrome (MDS), to evaluate its impact on outcome and its possible association with other known prognostic factors. MATERIAL/METHODS: The CD200 was detected by flow cytometry, and the chromosome karyotypes were determined by G banding respectively. The Mann-Whitney U test was used to analyze the association among CD200 expression and clinical features. In addition, the overall survival and AML transformation of the MDS patients according to the expression level of CD200 was also explored. RESULTS: Overall, the flow cytometric analyses confirmed that expression of CD200 was high in this patient cohort compared to normal BM (p<0.01). The levels of CD200 in RCUD (20.3%±4.3%), RCMD (25.0%±4.5%), RAEB-1 (39.2%±4.9%), and RAEB-2 (43.2%±5.8%) groups were obviously higher than that of RARS group (6.8%±1.7%, P<0.05). Significant differences of CD200 expression were observed in the 4 groups of MDS according to IPSS risk(P<0.01). After 45-month follow-up, Kaplan-Meier analysis of patients with MDS in our study indicated that patients with high expression level of CD200 had a shorter overall survival and a high Leukemic transformation than those with low expression (p<0.01). CONCLUSIONS: In conclusion, our findings provide firstly the evidence that CD200 is up-regulated and emerging as both a prognostic factor and a potential target of novel therapeutic approaches for MDS.
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Antígenos CD/genética , Síndromes Mielodisplásicos/diagnóstico , Adulto , Anciano , Anciano de 80 o más Años , Transformación Celular Neoplásica , Femenino , Citometría de Flujo , Humanos , Cariotipificación , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/clasificación , Síndromes Mielodisplásicos/genética , Síndromes Mielodisplásicos/mortalidad , Pronóstico , Tasa de Supervivencia , Regulación hacia ArribaRESUMEN
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OBJECTIVE: To investigate the effects of anti-CD44 mAb A3D8 on the cell proliferation of human acute monocytic leukemia cell line THP-1 and its mechanism. METHODS: Cell proliferation was assayed with MTT method, the expression of CD33, CD15, CD11b, CD14, Annexin-V, caspase-3 and cell cycle with flow cytometry, and the expression of p-Akt, p-ERK, bcl-2 and p27kip1 with Western blot. RESULTS: A3D8 could remarkably inhibit the proliferation capacity of the THP-1 cells in a dosage- and time-dependent manner. THP-1 differentiation was observed when treated with A3D8 (2.0 µg/ml) for one to six days. Expression of CD33 (68.9 ± 2.0 vs 39.3 ± 1.5), CD15 (61.7 ± 5.5 vs 12.9 ± 2.6), CD11b (67.3 ± 3.8 vs 14.0 ± 2.0) and CD14 (83.0 ± 5.7 vs 8.0 ± 1.0) was significantly increased at day 4 compared with the control group (all P < 0.01). Cell cycle of the THP-1 cells was arrested in G(0)/G(1). Expression of the Annexin-V \[(32.5 ± 2.5)% vs (2.4 ± 0.3)%\] and caspase-3 \[(33.3 ± 2.5)% vs (3.6 ± 0.3)%\] was much higher than that in normal controls (all P < 0.01), and apoptosis was observed in THP-1 cells at day 5. Expression of p-Akt (0.24 ± 0.06 vs 1.20 ± 0.15), p-ERK (0.32 ± 0.05 vs 1.24 ± 0.09), and bcl-2 (0.11 ± 0.05 vs 0.65 ± 0.07) was much lower than that of the controls (all P < 0.01), while p27kip1 (1.08 ± 0.09 vs 0.10 ± 0.02) was significantly increased at day 4 (P < 0.05). CONCLUSION: Anti-CD44 antibody can induce the differentiation and apoptosis of THP-1 cell through inhibiting PI3K/AKt and ERK1/2 signaling pathway.
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Anticuerpos Monoclonales/farmacología , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Anticuerpos Monoclonales/inmunología , Línea Celular Tumoral , Humanos , Receptores de Hialuranos/inmunología , Leucemia Monocítica Aguda/patología , Transducción de SeñalRESUMEN
OBJECTIVE: To investigate the expression of CD123 and its significance in lymphocytic leukemia. METHODS: CD123 expression in 139 lymphocytic leukemia patients and in lymphocytes from 10 normal bone marrows (BM) was analyzed by multi-parameter flow cytometry. Cytogenetic and minimal residual disease (MRD) analysis were performed in acute B-lymphocytic leukemia (B-ALL) patients. RESULTS: CD123 expression was absent in B lymphoid lineage stem-progenitor cells, mature B and T lymphocytes from 10 normal BM. Among 139 lymphocytic leukemia patients, CD123 was negative in 5 T-ALL and 23 B-CLL patients. However, among 111 B-ALL patients, CD123 was expressed in 106 (12 pro B-ALL, 57 common B-ALL and 37 Pre B-ALL) (95.49%) but not in 5 mature B-ALL patients. There was a positive correlation between CD123 and p-Akt expression, and CD123 expression was much higher in hyperdiploid than in non-hyperdiploid B-ALL patients. A statistically significant difference in relapse rate within 12 months (MRD positive group: 63.04% vs MRD negative group 21.56%)and in disease free survival (DFS) time was found beween patients with MRD\[(36.06 +/- 2.62)%\] or not \[(48.23 +/- 1.82)%\] (P < 0.01). Moreover, stable CD123 expression could be observed in B-ALL patients in relapse. CONCLUSIONS: CD123 was predominantly expressed in B-ALL patients and remained in patients in relapsec, indicating that it may be an useful MRD marker in B-ALL patients.
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Neoplasia Residual , Leucemia-Linfoma Linfoblástico de Células Precursoras B , Citometría de Flujo , Humanos , Leucemia Linfocítica Crónica de Células B , Leucemia-Linfoma Linfoblástico de Células T PrecursorasRESUMEN
The aim of this study was to investigate the effect of rapamycin on cell growth and apoptosis in the myelodysplastic syndrome (MDS) cell line MUTZ-1 and possible mechanism. MUTZ-1 cells were treated with rapamycin, cell proliferation capability was determined with MTT, protein expression including Annexin V/PI, caspase 3, PTEN, p-Akt, p-mTOR and the cell cycle were analyzed with flow cytometry. The results indicated that the proliferation of MUTZ-1 cells was inhibited by rapamycin in concentration-and time-dependent manners (r=0.67, 0.61, 0.72). After treatment with rapamycin for 24-72 hours, cell count in G0/G1 were significantly higher than that of the control (p<0.01), and this effect showed a time-and concentration-dependency (r=0.94, 0.93, 0.92), the cell cycle was blocked in G0/G1 phase. As compared with control group, the proportion of Annexin V+PI-MUTZ-1 cells and the cellular PTEN levels increased in the treated group dramatically and in time-and dose-dependent manners (p<0.01). To the contrary, level of p-mTOR expression markedly decreased as compared with control group (p<0.05). It is concluded that the rapamycin inhibits the proliferation of MUTZ-1 cells, down-regulates the PTEN/PI3K-Akt/mTOR signaling pathway by interaction with mTOR, which induces the apoptosis of mUTZ-1 cells.
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Apoptosis/efectos de los fármacos , Síndromes Mielodisplásicos/patología , Transducción de Señal/efectos de los fármacos , Sirolimus/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica , Humanos , Síndromes Mielodisplásicos/metabolismo , Serina-Treonina Quinasas TOR/metabolismoRESUMEN
OBJECTIVE: To investigate the relationship between PTEN gene expression and Akt phosphorylation (p-Akt) in myelodysplastic syndrome (MDS) and to explore the progression of MDS and the mechanism of high risk transformation to acute myeloid leukemia. METHODS: RT-PCR was used to detect the PTEN mRNA expression in leukemia cell lines K562 (as negative control) and Jurkat (as positive control) and 65 MDS and MDS/AML patients. Flow cytometry was used to detect p-Akt in HL-60 and Jurkat cells and 30 MDS patients. RESULTS: (1) K562 cells present PTEN gene expression while Jurkat cells did not. Of 65 MDS and MDS/AML patients, 27 (41.5%) expressed PTEN mRNA, being significantly lower than that in normal group (85.7%) (P < 0.01). (2) Jurkat cell showed high expression (86.9%) of p-Akt, while HL-60 cell as negative control did not express. P-Akt levels of 30 MDS patients were increased (1.35% - 58.23%), being much higher as compared with that of the normal contrast group (0.54% - 2.34%) (P < 0.01). Moreover, with the rate of blast cells increasing, the p-Akt level was rising up. There is a positive correlation (r = 0.93, P < 0.01) between the low expression rate of PTEN and the positive rate of p-Akt. CONCLUSION: The loss of PTEN gene expression is one of the important factors of p-Akt high expression in MDS patients, moreover, it may speed up the progress of the MDS or transformation to acute myeloid leukemia.
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Síndromes Mielodisplásicos/metabolismo , Fosfohidrolasa PTEN/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Adulto , Anciano , Células de la Médula Ósea/metabolismo , Femenino , Células HL-60 , Humanos , Células Jurkat , Células K562 , Masculino , Persona de Mediana Edad , Fosforilación , ARN Mensajero/metabolismoRESUMEN
To evaluate the sensitivity and specificity analysis of the lineage related antibodies in acute leukemia immunophenotyping by flow cytometry (FCM), immunophenotyping in 184 patients with acute leukemia was performed by FCM analysis. The results showed that in the lineage-related antibodies of acute myelocytic leukemia (AML), the sensitivity of CD13 and CD33 was higher (95.5% and 91.2%, respectively), the specificity of them was deficient (72.5% and 62.2%, respectively); the sensitivity of MPO was low (69.1%), but the specificity was high (100%); the sensitivity and specificity of CD117 were high (88.2% and 100%, respectively); the sensitivity of CD14 and CD15 was low (18.4% and 27.2%, respectively); the specificity of CD14 with monocytes was high. As the lineage-related antibodies of B-lineage ALL were concerned, CD19 showed high sensitivity and low specificity (100% vs 83.4%); the sensitivity and specificity of CD79a (96.4% vs 100%) and CD22 (100% vs 100%) were high; the sensitivity and specificity of CD10 (53.6% vs 82.5%) and CD20 (70.4% vs 87.5%) were low. In T-lineage ALL, the specificity of CD3 was high (97.5%), but the sensitivity was below the mark (80.0%); the sensitivity of CD7 was high (100%), but the specificity was low (77.9%); while the sensitivity and specificity of CD5, CD2 and CD1a were all deficient. In conclusion, the sensitivity and specificity analysis of the lineage-related antibodies in acute leukemia immunophenotyping are coincident with St Jude immunophenotyping project. It seems only that CD117 is superior to MPO in defining AML, but the sensitivity and specificity analysis of CD22 and CD79 are similar in defining B-lineage ALL, therefore, anyone of them may be selected as your need.
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Citometría de Flujo/métodos , Inmunofenotipificación/métodos , Leucemia/inmunología , Enfermedad Aguda , Adolescente , Adulto , Anciano , Anticuerpos Monoclonales/inmunología , Anticuerpos Antineoplásicos/inmunología , Antígenos CD79/análisis , Antígenos CD79/inmunología , Niño , Preescolar , Femenino , Humanos , Leucemia/clasificación , Leucemia Eritroblástica Aguda/inmunología , Leucemia Monocítica Aguda/inmunología , Leucemia Mieloide/inmunología , Leucemia Mieloide Aguda/inmunología , Leucemia Promielocítica Aguda/inmunología , Masculino , Persona de Mediana Edad , Proteínas Proto-Oncogénicas c-kit/análisis , Proteínas Proto-Oncogénicas c-kit/inmunología , Reproducibilidad de los Resultados , Lectina 2 Similar a Ig de Unión al Ácido Siálico/análisis , Lectina 2 Similar a Ig de Unión al Ácido Siálico/inmunologíaRESUMEN
To investigate the changes of platelet activated state and platelet activated function by trace whole blood flow cytometry (FCM), and to explore the mechanism of hemorrhage and infiltration in adults with acute leukemia, the expression percentage and changes of these expressions of CD62p and PAC-1 on platelet surface were determined by FCM of trace whole blood after platelet activated by ADP in patients with new diagnosed AL (group I), complete remission (CR, group II) and continuously complete remission (CCR, group III). Healthy adults were used as control group. The result showed that the expression of CD62p in group I and II was higher than that in control group, before and after platelet activated by ADP (P < 0.01). The expression of PAC-1 in group I was higher than that in control group (P < 0.01), the expression of PAC-1 in group II was lower than that in control group (P > 0.01), There was no significant difference in expression of CD62p and PAC-1 between group III and control group (P > 0.01), and no significant difference was found between AL group with megakaryocyte malignant pathological changes and AL group without megakaryocyte malignant pathological changes before platelet activated by ADP (P > 0.01). After platelet activated by ADP, the expression of PAC-1 in the former was lower than that in the latter (P < 0.01). It is concluded that (1) high level activated platelet in peripheral blood of AL patients show that interaction between activated platelet and leukemia cells can be one of reason resulting in widespread hemorrhage and infiltration AL patiens; (2) the decrease of number and activted function of platelet at the first stage of AL patients may be caused by malignant hyperplasia of leukemia cells and damage of megakaryopoiesis in bone marrow.
Asunto(s)
Plaquetas/metabolismo , Leucemia/sangre , Activación Plaquetaria/fisiología , Enfermedad Aguda , Adenosina Difosfato/farmacología , Adolescente , Adulto , Anciano , Plaquetas/citología , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Femenino , Citometría de Flujo , Humanos , Leucemia/patología , Masculino , Persona de Mediana Edad , Selectina-P/biosíntesis , Activación Plaquetaria/efectos de los fármacos , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/biosíntesisRESUMEN
In order to study the influence of trichosanthin (TCS) on apoptosis and growth inhibition of human NB4 cells in vitro, the expression of annexin V and the change of DeltaPsim of NB4 cells induced by TCS was analyzed by FACS, and MTT assay was adopted to measure the growth inhibition ratio of NB4 cells treated with TCS. Apoptosis was assayed by agarose gel electrophoresis. The results showed the higher concentration of TCS and the longer the acting time, the stronger growth inhibition of NB4 cells. The expression of annexin V was positive, and the positive ratio was greatly enhanced with prolongation of acting time. DeltaPsim reduced gradually while the apoptosis cells increasing. DNA agarose gel electrophoresis showed a gradient, which confirmed that TCS could induce NB4 cells apoptosis. In conclusion, taken together, data show that TCS can inhibit NB4 growth in vitro, and induce apoptosis. Experiment provides an important evidence for application of TCS in clinical treatment of acute promyelocytic leukemia.