human leukemic monocyte lymphoma cell line u937  (ATCC)

Journal: Frontiers in Oncology

Article Title: Curcumin in Combination With Omacetaxine Suppress Lymphoma Cell Growth, Migration, Invasion, and Angiogenesis via Inhibition of VEGF/Akt Signaling Pathway

doi: 10.3389/fonc.2021.656045

Figure Lengend Snippet: Combination of curcumin with HHT inhibited the proliferation of lymphoma cells. U937 and Raji cells were treated with 5 ng/mL HHT or/and 10 μM curcumin for 72 h. (A) CCK-8 assay was used to detect cell viability. (B) Relative fluorescence expression levels were quantified by Ki67 and DAPI staining. **P < 0.01 compared with control group; ## P < 0.01 compared with HHT group.

Article Snippet: Human umbilical vein endothelial cells (HUVECs), human leukemic monocyte lymphoma cell line U937, Burkitt’s lymphoma cell line Raji and human bone marrow stromal cell line HS-5 were purchased form American Type Culture Collection (ATCC, Rockville, Maryland, USA).

Techniques: CCK-8 Assay, Fluorescence, Expressing, Staining, Control

Journal: Frontiers in Oncology

Article Title: Curcumin in Combination With Omacetaxine Suppress Lymphoma Cell Growth, Migration, Invasion, and Angiogenesis via Inhibition of VEGF/Akt Signaling Pathway

doi: 10.3389/fonc.2021.656045

Figure Lengend Snippet: Combination of curcumin with HHT suppressed the migration and invasion abilities of lymphoma cells. U937 and Raji cells were treated with 5 ng/mL HHT or/and 10 μM curcumin, or treated with HHT, curcumin and VEGF for 24 h. (A, B) Cell migration was detected by transwell migration assay. (C, D) Cell invasion was detected by transwell invasion assay. *P < 0.05, **P < 0.01 compared with control group; ## P < 0.01 compared with HHT group; ^^ P < 0.01 compared with HHT + curcumin group.

Article Snippet: Human umbilical vein endothelial cells (HUVECs), human leukemic monocyte lymphoma cell line U937, Burkitt’s lymphoma cell line Raji and human bone marrow stromal cell line HS-5 were purchased form American Type Culture Collection (ATCC, Rockville, Maryland, USA).

Techniques: Migration, Transwell Migration Assay, Transwell Invasion Assay, Control

Journal: Current Issues in Molecular Biology

Article Title: Hesperetin Induces Autophagy and Delayed Apoptosis by Modulating the AMPK/Akt/mTOR Pathway in Human Leukemia Cells In Vitro

doi: 10.3390/cimb45020102

Figure Lengend Snippet: Effect of hesperetin on human leukemic cell viability. HL-60 and U937 cell lines (4 × 10 4 cells/mL) were treated with hesperetin at 0, 12.5, 25, 50, and 100 µM for 24 h ( A ) and 48 h ( B ). Cell viability was evaluated using a CCK-8 assay. The Y -axis indicates the percentage of cell survival, and the X -axis indicates various concentrations of hesperetin. The mean ± SD of the three independent experiments performed in triplicate are shown. * p < 0.05; ** p < 0.01; *** p < 0.001 vs. the control group; ns, not significant; the results are representative of three independent experiments.

Article Snippet: The human leukemic monocyte lymphoma cell line U937 and the promyelocytic leukemia cell line HL-60 were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan; derived from ATCC CRL-1593.2 and ATCC CCL-240).

Techniques: CCK-8 Assay, Control

Journal: Current Issues in Molecular Biology

Article Title: Hesperetin Induces Autophagy and Delayed Apoptosis by Modulating the AMPK/Akt/mTOR Pathway in Human Leukemia Cells In Vitro

doi: 10.3390/cimb45020102

Figure Lengend Snippet: Effects of hesperetin compounds on apoptosis and cell cycle arrest of U937 cells. ( A ) U937 cell apoptosis was analyzed at 24 and 48 h by flow cytometry with annexin V-FITC/PI staining to distinguish early apoptotic (annexin V-FITC positive, PI negative; Q4-1 and Q4) from late apoptotic or necrotic cells (Annexin V-FITC positive, PI positive; Q2-1 and Q2). ( B ) The cell cycle was assessed using flow cytometry in U937 cells with or without hesperetin treatment. ( C ) U937 cells were treated with hesperetin at 0, 12.5, 25, 50, and 100 µM for 48 h. Cleaved-PARP-1, Bcl-2, Bax, and GAPDH expressions were analyzed with Western blotting by using the cell lysates. The mean ± SD of the three independent experiments performed in triplicate are shown. * p < 0.05; ** p < 0.01; *** p < 0.001 vs. the control group; ns, not significant; the results are representative of three independent experiments.

Article Snippet: The human leukemic monocyte lymphoma cell line U937 and the promyelocytic leukemia cell line HL-60 were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan; derived from ATCC CRL-1593.2 and ATCC CCL-240).

Techniques: Flow Cytometry, Staining, Western Blot, Control

Journal: Current Issues in Molecular Biology

Article Title: Hesperetin Induces Autophagy and Delayed Apoptosis by Modulating the AMPK/Akt/mTOR Pathway in Human Leukemia Cells In Vitro

doi: 10.3390/cimb45020102

Figure Lengend Snippet: Effects of hesperetin on autophagy in U937 cells. ( A ) To evaluate the change in the number of AVOs in cells treated with hesperetin, the cells were treated with different concentrations of hesperetin at 24 and 48 h. Next, the cells were stained with acridine orange (1 μg/mL) at 37 °C for 15 min in the dark. The cells were analyzed using a FACScan flow cytometer. The data were analyzed using BD Cell Quest software. ( B ) LC3-I/II, Beclin-1, Atg5, p62, and GAPDH protein expression for 24 and 48 h ( C ) were analyzed with Western and semiquantified in representative results of the same pattern from three independent experiments are shown. ( D ) U937 cells were exposed to the indicated concentration of hesperetin in the presence or absence of autophagy inhibitor 3-MA (left panel) or Baf-A1 (right panel) for 24 h. Cell viability was assessed using the CCK-8 assay. The mean ± SD of the three independent experiments performed in triplicate are shown. * p < 0.05; ** p < 0.01; *** p < 0.001 vs. the control group; ns, not significant; the results are representative of three independent experiments.

Article Snippet: The human leukemic monocyte lymphoma cell line U937 and the promyelocytic leukemia cell line HL-60 were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan; derived from ATCC CRL-1593.2 and ATCC CCL-240).

Techniques: Staining, Flow Cytometry, Software, Expressing, Western Blot, Concentration Assay, CCK-8 Assay, Control

Journal: Current Issues in Molecular Biology

Article Title: Hesperetin Induces Autophagy and Delayed Apoptosis by Modulating the AMPK/Akt/mTOR Pathway in Human Leukemia Cells In Vitro

doi: 10.3390/cimb45020102

Figure Lengend Snippet: Effect of hesperetin on the AMPK/Akt/mTOR signaling pathway in U937 cells. U937 cells (5 × 10 5 ) were treated with hesperetin at 0, 12.5, 25, 50, and 100 μM for 24 h ( A ) and 48 h ( B ). The expression of phospho-AMPK Thr172 , AMPK, phospho-mTOR Ser2448 , mTOR, Phospho-Akt Ser473 , Akt and GAPDH was analyzed with Western blotting and semiquantified; representative results of the same pattern from three independent experiments are shown. The mean ± SD of the three independent experiments performed in triplicate are shown. * p < 0.05; ** p < 0.01; *** p < 0.001 vs. the control group; ns, not significant; the results are representative of three independent experiments.

Article Snippet: The human leukemic monocyte lymphoma cell line U937 and the promyelocytic leukemia cell line HL-60 were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan; derived from ATCC CRL-1593.2 and ATCC CCL-240).

Techniques: Expressing, Western Blot, Control

Journal: Current Issues in Molecular Biology

Article Title: Hesperetin Induces Autophagy and Delayed Apoptosis by Modulating the AMPK/Akt/mTOR Pathway in Human Leukemia Cells In Vitro

doi: 10.3390/cimb45020102

Figure Lengend Snippet: Schematic diagram of hesperetin molecular mechanism in leukemia cells. Hesperetin modulates AMPK/Akt/mTOR signaling and induces autophagy and delayed apoptosis by regulating the AMPK/Akt/mTOR pathway through AMPK activation, Akt and mTOR downregulation, inactivating and activating various target proteins, such as cleaved-PARP-1, Bcl-2, Bax, LC3-I/II, Beclin-1, Atg5, and p62; hesperetin promoted cell death in the human leukemic cell line U937 by inducing a low degree of slight apoptosis, cell cycle arrest, and autophagy, which increases the anticancer effect on leukemia (created with BioRender.com).

Article Snippet: The human leukemic monocyte lymphoma cell line U937 and the promyelocytic leukemia cell line HL-60 were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan; derived from ATCC CRL-1593.2 and ATCC CCL-240).

Techniques: Activation Assay

Journal: Oncotarget

Article Title: xCT increases tuberculosis susceptibility by regulating antimicrobial function and inflammation

doi: 10.18632/oncotarget.9052

Figure Lengend Snippet: ( A ) xct expression in macrophages infected with H37Ra (MOI = 5), H37Rv (MOI = 5) or stimulated with Mtb lysates 10 μg/ml, 19 kD protein (1 μg/ml), LPS (100 ng/ml). ( B ) xCT expression in macrophage treated without or with anti-TLR2 mAb in the absence or presence of Mtb lysate stimulation for 24 hours. ( C ) qPCR detection of xCT in U937 macrophage infected with Mtb at MOI of 5 for 24 h in the presence of different kinase inhibitors, SB203580 (10 μM), SP600125 (10 μM), U0126 (10 μM), LY294002 (10 μM), BAY117082 (10 μM). ( D ) Western blot analysis of xCT expression in U937 macrophage infected with Mtb at MOI of 5 for 24 h in the presence of different kinase inhibitors, SB203580 (10 μM), LY294002 (10 μM). ( E ) Western blot of the signaling pathways, p38 and Akt, in U937 macrophage infected with Mtb at MOI of 5 for indicated time. Representative data of 3 experiments were expressed as mean ± SEM * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

Article Snippet: Human leukemic monocyte lymphoma cell line U937 cells (ATCC CRL 1593), were cultured in RPMI1640, supplemented with 10% fatal bovine serum (FBS), antibiotics (50 U/ml penicillin and 50 μg/ml streptomycin), 0.1 mM non-essential amino acids and 1 mM sodium pyruvate in 5% CO 2 at 37°C.

Techniques: Expressing, Infection, Western Blot, Protein-Protein interactions

Journal: Oncotarget

Article Title: xCT increases tuberculosis susceptibility by regulating antimicrobial function and inflammation

doi: 10.18632/oncotarget.9052

Figure Lengend Snippet: ( A, C, E ) Peritoneal macrophages from WT and xCT −/− mice were infected with Mtb at MOI of 5 in the absence or presence of NAC (500 μM). ( B, D, F ) U937 macrophages pretreated without or with SASP (200 μM), 4-CPG (100 μM) were infected with H37Ra at MOI of 5 in the absence or presence of NAC (500 μM). (A) CFUs in H37Ra or H37Rv infected macrophage lysates were enumerated at 3d after infection. (B) CFUs in H37Ra or H37Rv infected macrophage lysates were enumerated at 3d after infection. (C) The concentrations of intracellular GSH were evaluated by ELISA at 24 h after infection. (D) The concentrations of intracellular GSH were determined by ELISA at 24 h after infection. (E) CFUs in infected macrophage in the absence or presence of NAC (500 μM) were enumerated at 3 d after infection. (F) CFUs in infected macrophage in the absence or presence of NAC (500 μM) were enumerated at 3 d after infection. H37Ra survival rates were calculated by using mock or WT as a control. ( G, H ) Redox status of intracellular H37Rv population within macrophages. Immortalized C57BL/6 BMDM were infected with H37Rv that was engineered to express Mrx1-roGFP2, and ~30,000 cells were analyzed by flow cytometry by exciting at 405 and 488 nm lasers at a constant emission (510 nm). (G) FACS dot plot of infected macrophages with Mrx1-roGFP2-expressing H37Rv in the absence of SASP and presence of SASP at 48 h p.i. (H) Bar graph represents percentage of oxidized subpopulation of Mtb during different time points post macrophage infection. Values represent the mean ± S.E. of duplicate determinations of experiments repeated twice. Statistical significance was calculated using unpaired student's t test. Mean ± SD reported. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

Article Snippet: Human leukemic monocyte lymphoma cell line U937 cells (ATCC CRL 1593), were cultured in RPMI1640, supplemented with 10% fatal bovine serum (FBS), antibiotics (50 U/ml penicillin and 50 μg/ml streptomycin), 0.1 mM non-essential amino acids and 1 mM sodium pyruvate in 5% CO 2 at 37°C.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Control, Flow Cytometry, Expressing