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European Review For Medical and... Feb 2018To detect the expression of long non-coding RNA-CRNDE in patients with acute myeloid leukemia and its effect on proliferation and apoptosis in acute myeloid leukemia...
OBJECTIVE
To detect the expression of long non-coding RNA-CRNDE in patients with acute myeloid leukemia and its effect on proliferation and apoptosis in acute myeloid leukemia cell line U937.
PATIENTS AND METHODS
81 cases of newly diagnosed acute myeloid leukemia (AML) were enrolled, and 35 non-malignant hematological patients were selected as controls. Quantitative RT-PCR (qRT-PCR) was performed to detect the expression of lncRNA-CRNDE in the bone marrow specimens of the subjects, and the difference between the two groups was also compared. The correlation between the expression of lncRNA-CRNDE and the sex, age, classification and total survival of clinical patients was analyzed according to the clinical data. U937 cells and monocytes isolated from normal people were cultured, and the expression of lncRNA-CRNDE in acute myeloid leukemia cell line U937 and normal monocytes was compared. SiRNA-CRNDE and pcDNA-CRNDE were transfected into U937 cells, and cell counting kit-8 (CCK-8) assay was performed to detect proliferation of U937 cells, Annexin V/PI flow cytometry was carried out to detect cell apoptosis. Cell cycle was measured by flow cytometry.
RESULTS
The expression of lncRNA-CRNDE in patients with AML and U937 cells was significantly higher than that in non-malignant hematological controls. Results of clinical data showed that the expression of lncRNA-CRNDE was associated with the classification and total survival of myeloid leukemia in clinical patients. After transfection of siRNA-CRNDE, the proliferation and cloning ability of U937 cells decreased, while the apoptosis increased (p < 0.01) and cells were arrested in G0-G1 phase. Meanwhile, after transfection of pcDNA-CRNDE, the proliferation ability of U937 cells increased significantly, which indicated that the expression of lncRNA-CRNDE might play an essential role in promoting the proliferation of U937 cells.
CONCLUSIONS
LncRNA-CRNDE is highly expressed in the bone marrow tissues of AML patients, and the expression level is negatively correlated with the total survival of those clinical patients. Meanwhile, the expression is higher in FAB type M4 and M5 than that in M1, M2 and M3. LncRNA-CRNDE promotes the proliferation and cell cycle of U937 cells, and inhibits cell apoptosis, which is expected to become a molecular marker for predicting and treating AML.
Topics: Adolescent; Adult; Biomarkers, Tumor; Cell Line, Tumor; Cell Proliferation; Disease Progression; Female; Gene Expression Regulation, Neoplastic; Humans; Leukemia, Myeloid, Acute; Male; Middle Aged; RNA, Long Noncoding; Survival Rate; U937 Cells; Young Adult
PubMed: 29461608
DOI: 10.26355/eurrev_201802_14310 -
International Immunopharmacology Jun 2024Patients with diabetes are particularly susceptible to Legionella pneumophila (LP) infection, but the exact pathogenesis of LP infection in diabetic patients is still...
BACKGROUND
Patients with diabetes are particularly susceptible to Legionella pneumophila (LP) infection, but the exact pathogenesis of LP infection in diabetic patients is still not fully understood. Herein, we investigated the effect of diabetes on immune function during LP infection in vitro and in vivo.
METHODS
The time course of LP infection in macrophages under normal and high-glucose (HG) conditions was examined in vitro. Western blot was used to determine nucleotide-binding oligomerization domain 1 (NOD1), kinase 1/2 (ERK1/2), mitogen-activated protein kinase p38 (MAPK p38), and c-Jun N-terminal kinases (JNK). Enzyme-linked immunosorbent assay (ELISA) was used to assess the secretion of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Cell Counting Kit-8 (CCK8) assay assessed U937 cell viability after treating cells with different concentrations of high sugar medium and ML130 (NOD1 inhibitor). For the in vivo study, normal and streptozocin-induced diabetic guinea pigs were infected with LP for 6, 24, and 72 h, after which NOD1, MAPK-related signals, TNF-α, and IL-6 expression in lung tissues were assessed using immunohistochemistry, western blot, and RT-PCR.
RESULTS
HG attenuated the upregulation of NOD1 expression and reduced TNF-α and IL-6 secretion caused by LP compared with LP-infected cells exposed to normal glucose levels (all p < 0.05). In diabetic guinea pigs, HG inhibited the upregulation of NOD1 expression in lung tissues and the activation of p38, ERK1/2, and cJNK caused by LP infection compared to control pigs (all p < 0.05).
CONCLUSION
HG attenuates the response of macrophages to LP infection by inhibiting NOD1 upregulation and the activation of MAPK signaling.
Topics: Nod1 Signaling Adaptor Protein; Animals; Humans; Macrophages; Legionella pneumophila; Glucose; Guinea Pigs; Male; Interleukin-6; Legionnaires' Disease; Diabetes Mellitus, Experimental; MAP Kinase Signaling System; U937 Cells; Tumor Necrosis Factor-alpha; Mice
PubMed: 38749333
DOI: 10.1016/j.intimp.2024.112254 -
Pulsating electromagnetic field stimulation prevents cell death of puromycin treated U937 cell line.Journal of Physiology and Pharmacology... Apr 2010Aim of study was to verify whether pulsating electromagnetic field (PEMF) can affect cancer cells proliferation and death. U937 human lymphoid cell line at densities...
Aim of study was to verify whether pulsating electromagnetic field (PEMF) can affect cancer cells proliferation and death. U937 human lymphoid cell line at densities starting from 1 x 10(6) cells/ml to 0.0625 x 10(6) cells/ml, were exposed to a pulsating magnetic field 50 Hz, 45+/-5 mT three times for 3 h per each stimulation with 24 h intervals. Proliferation has been studied by counting number of cells stimulated and non-stimulated by PEMF during four days of cultivation. Viability of cells was analyzed by APC labeled Annexin V and 7-AAD (7-amino-actinomycin D) dye binding and flow cytometry. Growing densities of cells increase cell death in cultures of U937 cells. PEMF exposition decreased amount of cells only in higher densities. Measurement of Annexin V binding and 7-AAD dye incorporation has shown that density-induced cell death corresponds with decrease of proliferation activity. PEMF potentiated density-induced death both apoptosis and necrosis. The strongest influence of PEMF has been found for 1 x 10(6)cells/ml and 0.5 x 10(6) cells/ml density. To eliminate density effect on cell death, for further studies density 0.25 x 10(6) cells/ml was chosen. Puromycin, a telomerase inhibitor, was used as a cell death inducer at concentration 100 microg/ml. Combined interaction of three doses of puromycin and three fold PEMF interaction resulted in a reduced of apoptosis by 24,7% and necrosis by 13%. PEMF protects U937 cells against puromycin- induced cell death. PEMF effects on the human lymphoid cell line depends upon cell density. Increased density induced cells death and on the other hand prevented cells death induced by puromycin.
Topics: Antimetabolites, Antineoplastic; Apoptosis; Cell Proliferation; Cell Survival; Electromagnetic Fields; Humans; Lymphoma, Large B-Cell, Diffuse; Necrosis; Puromycin; U937 Cells
PubMed: 20436221
DOI: No ID Found -
Ultrasonics Sonochemistry Mar 2009Three novel lipid-shell-type microbubbles (MBs), AS-0100, BG6356A and BG6356B, have been evaluated for their impact on ultrasound (US)-induced cell death and free... (Comparative Study)
Comparative Study
Three novel lipid-shell-type microbubbles (MBs), AS-0100, BG6356A and BG6356B, have been evaluated for their impact on ultrasound (US)-induced cell death and free radicals production. Previously studied and well-characterized US exposure conditions were employed in which human myelomonocytic lymphoma U937 cells were exposed to 1MHz pulsed US beam (0.3W/cm(2), 10% duty factor) for 1min with or without MBs. Three different concentrations of each MB were used. Apoptosis and cell lysis were assessed by examining phosphatidylserine externalization and by counting viable cells, respectively, 6h post-exposure. Free radicals production and scavenging activities were evaluated using electron paramagnetic resonance (EPR)-spin trapping. The results showed that only AS-0100 and BG6356A were able to enhance the US-induced apoptosis, mainly by increasing the secondary necrosis. Apoptosis and cell lysis seemed to depend more on mechanical forces exerted by oscillating MBs while free radicals played a trivial role. BG series MBs exhibited pronounced scavenging activities. Generally, despite the need for further optimization, AS-0100 and BG6356A appear to be promising as adjuncts in cases where US-induced cell death is required.
Topics: Cell Death; Contrast Media; Free Radicals; Humans; Microbubbles; Sonication; U937 Cells
PubMed: 19014893
DOI: 10.1016/j.ultsonch.2008.10.003 -
Cancer Research Nov 2006Stromal cell-derived factor-1 (SDF-1/CXCL12) and its receptor CXCR4 are implicated in the pathogenesis and prognosis of acute myelogenous leukemia (AML). Cellular...
Stromal cell-derived factor-1 (SDF-1/CXCL12) and its receptor CXCR4 are implicated in the pathogenesis and prognosis of acute myelogenous leukemia (AML). Cellular microparticles, submicron vesicles shed from the plasma membrane of various cells, are also associated with human pathology. In the present study, we investigated the putative relationships between the SDF-1/CXCR4 axis and microparticles in AML. We detected CXCR4-expressing microparticles (CXCR4(+) microparticles) in the peripheral blood and bone marrow plasma samples of normal donors and newly diagnosed adult AML patients. In samples from AML patients, levels of CXCR4(+) microparticles and total SDF-1 were elevated compared with normal individuals. The majority of CXCR4(+) microparticles in AML patients were CD45(+), whereas in normal individuals, they were mostly CD41(+). Importantly, we found a strong correlation between the levels of CXCR4(+) microparticle and WBC count in the peripheral blood and bone marrow plasma obtained from the AML patients. Of interest, levels of functional, noncleaved SDF-1 were reduced in these patients compared with normal individuals and also strongly correlated with the WBC count. Furthermore, our data indicate NH(2)-terminal truncation of the CXCR4 molecule in the microparticles of AML patients. However, such microparticles were capable of transferring the CXCR4 molecule to AML-derived HL-60 cells, enhancing their migration to SDF-1 in vitro and increasing their homing to the bone marrow of irradiated NOD/SCID/beta2m(null) mice. The CXCR4 antagonist AMD3100 reduced these effects. Our findings suggest that functional CXCR4(+) microparticles and SDF-1 are involved in the progression of AML. We propose that their levels are potentially valuable as an additional diagnostic AML variable.
Topics: Adult; Aged; Aged, 80 and over; Bone Marrow; Chemokine CXCL12; Chemokines, CXC; Female; HL-60 Cells; Humans; Leukemia, Myeloid, Acute; Leukocyte Count; Male; Middle Aged; Receptors, CXCR4; U937 Cells
PubMed: 17108140
DOI: 10.1158/0008-5472.CAN-06-2006 -
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi =... Nov 2021Objective To investigate the effect of lncRNA CRNDE on proliferation, apoptosis, and cell cycle of U937 cells and its mechanism. Methods The expression level of CRNDE in...
Objective To investigate the effect of lncRNA CRNDE on proliferation, apoptosis, and cell cycle of U937 cells and its mechanism. Methods The expression level of CRNDE in bone marrow cells of AML patients was analyzed by GEPIA database; the mRNA expression levels of miR-136-5p, CRNDE, and minichromosome maintenance 5(MCM5) in AML cell lines were detected by quantitative real-time PCR (qRT-PCR). The lentiviral vector with CRNDE knocked down was constructed and transfected into U937 cells which were randomized into CRNDE knockdown group (sh-CRNDE group) and negative control group (sh-NC group); miR-136-5p mimic and miR-136-5p inhibitor were transfected respectively to overexpress and knock down miR-136-5p in U937 cells which were randomized into miR-136-5p-mimic group, NC-mimic group, miR-136-5p-inhibitor group, and NC-inhibitor group. The effect of CRNDE and miR-136-5p on proliferation was detected by CCK-8 assay and cell counting assay, and the effect of them on cell cycle and apoptosis was detected by flow cytometry. The mRNA expressions of miR-136-5p, CRNDE, and MCM5 were detected by qRT-PCR, and the protein expressions of MCM5, Bcl2, cyclin D1, and cyclin A2 were detected by Western blotting. Results CRNDE was highly expressed in the bone marrow and cell lines of AML patients. Knockdown of CRNDE upregulated miR-136-5p, inhibited the MCM5 mRNA and protein expressions and the cell proliferation, promoted the cell apoptosis, and blocked the cell cycle in G1 phase. Overexpression of miR-136-5p also inhibited the expression of MCM5 at both mRNA and protein levels, while knockdown of miR-136-5p reversed those effects. Conclusion CRNDE promotes the proliferation and inhibits the apoptosis of U937 cells by downregulating miR-136-5p and upregulating MCM5.
Topics: Apoptosis; Cell Cycle Proteins; Cell Line, Tumor; Cell Proliferation; Humans; MicroRNAs; RNA, Long Noncoding; U937 Cells
PubMed: 34809738
DOI: No ID Found -
Clinical and Experimental Medicine May 2013Biomarkers on malignant cells may confer prognostic significance. Krüppel-like factor 4 (KLF4) has been reported to be expressed variably on leukemia cells, but its...
Biomarkers on malignant cells may confer prognostic significance. Krüppel-like factor 4 (KLF4) has been reported to be expressed variably on leukemia cells, but its expression patterns in leukemias with different differentiation stages and its relationships with clinical parameters remain to be elucidated. To examine the KLF4 expression pattern in human leukemias and its clinical significance, RT-PCR and real-time PCR were used to detect KLF4 expression in 9 leukemia cell lines and 96 pediatric leukemia patients. KLF4 mRNA expressed in 5/16 (31.25 %) of AML patients and 11/28 (39.29 %) ALL patients, significantly lower than that in control (9/11, 81.82 %, P < 0.05). The expressions of KLF4 mRNA were much lower in ALL (27/52) compared to AML (25/52) (P = 0.019), AML-M1 to other subtypes of AML (P = 0.001), C-ALL to Pre-B ALL (P = 0.004), Pro-T ALL to T-ALL (P = 0.048). Observation on leukemia cell lines showed the similar pattern. The relative expression of KLF4 mRNA was inversely associated with CD34-positive rates (r = -0.296, P = 0.037), but it was not associated with the blasts percentages in bone marrow (r = -0.222, P = 0.137) and the WBC counts (r = -0.058, P = 0.679). KLF4 mRNA expression level was not related to the overall survival (r = -0.063, P = 0.670), and the median survival times for the KLF4 (Lower) and KLF4 (Higher) groups were comparable (28 vs. 25 months, P = 0.265). Furthermore, no difference was found in KLF4 mRNA expression levels whether in leukemias with normal and abnormal karyotypes (P = 0.180), or in leukemias with normal and abnormal molecular cytogenetics (P = 0.591). We conclude that KLF4 translation level is associated with the differentiation stage of different leukemias and is independent of other parameters of risk stratification.
Topics: Adolescent; Antigens, CD34; Biomarkers, Tumor; Cell Differentiation; Child; Child, Preschool; Female; Gene Expression Regulation, Leukemic; Humans; Infant; K562 Cells; Kruppel-Like Factor 4; Kruppel-Like Transcription Factors; Leukemia; Male; Neoplasm Staging; Prognosis; Protein Biosynthesis; RNA, Messenger; RNA, Neoplasm; Severity of Illness Index; U937 Cells
PubMed: 22592916
DOI: 10.1007/s10238-012-0187-4 -
British Journal of Experimental... Oct 1979The deposition of quartz in the pulmonary alveoli creates a major demand for macrophages to replace those destroyed, but local proliferation of monocytes appeared to be...
The deposition of quartz in the pulmonary alveoli creates a major demand for macrophages to replace those destroyed, but local proliferation of monocytes appeared to be minimal and the role of systemic recruitment was therefore explored. Injected silica and lipids stimulated the phagocytic function of the mononuclear phagocytic system (MPS), whilst inhaled silica provoked lipid accumulation in the lung, thus suggesting that lipid might also induce a proliferative response in the marrow. Using marrow cultures, cells of the rat MPS were identified by size and phagocytic capacity for latex microspheres, and then subjected to kinetic analysis in litter-mate pairs by single and double labelling autoradiography, under normal conditions and after administration of lipid extracted from rat lungs consolidated by silica-induced alveolar lipo-proteinosis. Treatment of the results by a new device facilitated distinction of promonocytes from monocytes and thus afforded a more precise means of assessing MPS kinetics. The duration of DNA synthesis and the cell-cycle time of promonocytes were reduced and the rate of entry into DNA synthesis increased as a result of i.v. injection of lung lipid. These findings support the involvement of systemic recruitment of monocytes from the marrow by a positive feed-back mechanism when a powerful irritant persists in the lungs and the results are discussed in the overall context of silicotic fibrogenesis.
Topics: Animals; Bone Marrow Cells; Cell Count; Feedback; Kinetics; Lipids; Macrophages; Male; Mitosis; Phagocytosis; Pulmonary Alveoli; Rats; Silicosis
PubMed: 518823
DOI: No ID Found -
The Anatomical Record Nov 1997Milky spots in the human greater omentum are preformed specific accumulations of primarily macrophages within the stroma of the greater omentum. To obtain a better...
BACKGROUND
Milky spots in the human greater omentum are preformed specific accumulations of primarily macrophages within the stroma of the greater omentum. To obtain a better understanding of milky spots in the human greater omentum, the development and the earliest forms of milky spots in the human greater omentum were studied, with special attention to the macrophage population.
METHODS
Specimens of human greater omentum were obtained from fetuses of 20 to 40 weeks gestation and one newborn three days old (n = 6). Using mature macrophages (RFD 7), activated macrophages (RFD 1), B-lymphocytes (CD 22), and T-lymphocytes (CD 2), and immunoperoxydase labeling, the percentage of these cells in developing milky spots and the development of milky spots were studied by light microscopy. A time-dependent increase in the percentage of positive staining cells and the size of clusters was analyzed using the non-parametric Spearman rank correlation test.
RESULTS
Small accumulations of cells with about 50% monocytes/macrophages were present at 20 weeks of gestation. With increasing gestational age the number of clusters of cells increased significantly (P < 0.01) as well as their size (P < 0.01). Starting at 29 weeks, vascularized clusters of cells were seen; true milky spots were present at 35 weeks. A significant (P < 0.05) increase in the percentage of mature macrophages was found in developing milky spots, whereas no activated macrophages were seen. The percentage of B-lymphocytes and T-lymphocytes found in the clusters of cells and milky spots increased significantly (P < 0.05) but did not exceed 10% of the total number of cells.
CONCLUSIONS
From our data it can be concluded that milky spots are specific structures in the greater omentum formed between the 20th and 35th week of gestation. Further, we concluded that immature cells (promonocytes) mature locally in developing milky spots.
Topics: Cell Aggregation; Cell Count; Embryo, Mammalian; Embryonic and Fetal Development; Humans; Immunohistochemistry; Lymphocytes; Macrophages; Omentum; Staining and Labeling
PubMed: 9372174
DOI: 10.1002/(SICI)1097-0185(199711)249:3<399::AID-AR11>3.0.CO;2-J -
Molecular Medicine Reports Nov 2015Homeobox genes encode transcription factors that are essential for embryonic morphogenesis and differentiation. Transcription factors containing the highly conserved...
Homeobox genes encode transcription factors that are essential for embryonic morphogenesis and differentiation. Transcription factors containing the highly conserved homeobox motif show considerable promise as potential regulators of hematopoietic maturation events. Previous studies have suggested that the increased expression levels of homeobox (HOX)A genes was correlated with the cytogenetic findings associated with poor prognosis in acute myeloid leukemia and mixed lineage leukemia. The aim of the present study was to investigate the role of HOXA5 in leukemia. The U937 human leukemia cell line was transfected with a HOXA5‑targeted short hairpin RNA (shRNA) to determine the effects of downregulation of the HOXA5 on proliferation, apoptosis, cell cycle distribution and chemoresistance in leukemia cells. Reverse transcription‑quantitative polymerase chain reaction and western blot analyses demonstrated that the mRNA and protein expression levels of HOXA5 were markedly suppressed following transfection with an shRNA‑containing vector. Knockdown of HOXA5 significantly inhibited cell proliferation, as determined by Cell Counting kit‑8 assay. Flow cytometry revealed that reduced HOXA5 expression levels resulted in cell cycle arrest at the G1 phase, and induced apoptosis. In addition, western blot analysis demonstrated that HOXA5 knockdown increased the expression levels of caspase‑3, and reduced the expression levels of survivin in the U937 cells. Furthermore, knockdown of HOXA5 in the U937 cells enhanced their chemosensitivity to cytarabine. The results of the present study suggested that downregulation of HOXA5 by shRNA may trigger apoptosis and overcome drug resistance in leukemia cells. Therefore, HOXA5 may serve as a potential target for developing novel therapeutic strategies for leukemia.
Topics: Antimetabolites, Antineoplastic; Cell Proliferation; Cytarabine; Drug Resistance, Neoplasm; Homeodomain Proteins; Humans; Leukemia, Myeloid, Acute; RNA Interference; RNA, Small Interfering; U937 Cells
PubMed: 26397212
DOI: 10.3892/mmr.2015.4331