Journal: Frontiers in Oncology

Article Title: Daemonorops draco Blume Induces Apoptosis Against Acute Myeloid Leukemia Cells via Regulation of the miR-216b/c-Jun

doi: 10.3389/fonc.2022.808174

Figure Lengend Snippet: Daemonorops draco Blume (DD) exerted a cytotoxic effect on acute myeloid leukemia cells. The cytotoxicity of DD in (A) U937 and (B) THP-1 cells. The cells were treated with DD (i.e., 12.5, 25, 50, 100, or 200 μg/ml) for 24 h. Cell viability assay was performed using EZ-Cytox. Values above represent the means of three experiments. Means ± standard deviation; ** p < 0.01 and *** p < 0.001 compared with the untreated groups.

Article Snippet: The AML cell lines THP-1 and U937 were purchased from Korean Cell Line Bank (Seoul, South Korea).

Techniques: Viability Assay, Standard Deviation

Journal: Frontiers in Oncology

Article Title: Daemonorops draco Blume Induces Apoptosis Against Acute Myeloid Leukemia Cells via Regulation of the miR-216b/c-Jun

doi: 10.3389/fonc.2022.808174

Figure Lengend Snippet: Daemonorops draco Blume (DD) reduced mitochondrial membrane potential and induced apoptosis in U937 and THP-1 cells treated with DD. (A, B) Cells were pretreated at JC-1 (20 μM) and DD (15 and 30 μg/ml) for 4 h. Green monomeric fluorescence form changed to red fluorescent aggregates in a concentration-dependent manner, which was measured using a microplate reader. (C–F) After treatment with DD (15 and 30 μg/ml) for 24 h, the cells were subjected to Western blotting due to the expression of apoptosis-related proteins, such as cleaved caspase-3, cleaved poly(ADP-ribose)polymerase, CCAAT/enhancer-binding protein, p-ATF4, p-c-Jun, and β-actin, in (C, E) U937 and (D, F) THP-1 cells. Fluorescein isothiocyanate (excitation/emission = 540/570) and rhodamine (excitation/emission = 540/570). Values above represent the means of three experiments. Means ± standard deviation; * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the untreated groups.

Article Snippet: The AML cell lines THP-1 and U937 were purchased from Korean Cell Line Bank (Seoul, South Korea).

Techniques: Fluorescence, Concentration Assay, Western Blot, Expressing, Binding Assay, Standard Deviation

Journal: Frontiers in Oncology

Article Title: Daemonorops draco Blume Induces Apoptosis Against Acute Myeloid Leukemia Cells via Regulation of the miR-216b/c-Jun

doi: 10.3389/fonc.2022.808174

Figure Lengend Snippet: Daemonorops draco Blume (DD) increased DNA damage in U937 and THP-1 cells. (A, B) The effect of DD on H2A.X, which was treated with various concentrations of DD (i.e., 15 and 30 μg/ml for 24 h), in U937 and THP-1 cells, which were subjected to Western blotting with the antibodies of p-H2A.X and β-actin. (C, D) Confocal images of AML cells represented live cells (left panel), dead cells (middle panel), and the combination of both (right panel). AML cells were treated with DD (30 μg/ml) for 24 h, and double dyes were incubated at 37°C for 30 min. AML cells were stained with calcium AM (excitation/emission = 494/517) and ethidium homodimer-1 (excitation/emission = 528/617). Scale bar = 100 μm.

Article Snippet: The AML cell lines THP-1 and U937 were purchased from Korean Cell Line Bank (Seoul, South Korea).

Techniques: Western Blot, Incubation, Staining

Journal: Frontiers in Oncology

Article Title: Daemonorops draco Blume Induces Apoptosis Against Acute Myeloid Leukemia Cells via Regulation of the miR-216b/c-Jun

doi: 10.3389/fonc.2022.808174

Figure Lengend Snippet: Daemonorops draco Blume (DD) increased reactive oxygen species (ROS) accumulation and N-Acetyl-L-cysteine (NAC) pretreatment reversed the cytotoxic effect of DD in U937 and THP-1 cells. (A, B) Both cells were incubated with 20-μM 2’,7’-dichlorofluorescin diacetate (DCFDA) for 30 min at 37°C in the dark and subjected to ROS assay. The cells were exposed to 30-μg/ml DD for 4 h. DCFDA fluorescence was determined using a dual microplate reader. (C, D) Both cells were exposed to NAC (5 mM) pretreatment for 60 min and subjected to ROS measurement. (E, F) A cell viability assay was conducted with absorbance measurement using an optical spectrometer. (Ex/Em = 450/570). Values represent the means of three experiments. Means ± standard deviation; * p < 0.05, ** p < 0.01, and *** p < 0.001 compared to untreated control group. # p < 0.05 and ### p < 0.001 between the two groups.

Article Snippet: The AML cell lines THP-1 and U937 were purchased from Korean Cell Line Bank (Seoul, South Korea).

Techniques: Incubation, ROS Assay, Fluorescence, Viability Assay, Optical Spectrometry, Standard Deviation

Journal: Frontiers in Oncology

Article Title: Daemonorops draco Blume Induces Apoptosis Against Acute Myeloid Leukemia Cells via Regulation of the miR-216b/c-Jun

doi: 10.3389/fonc.2022.808174

Figure Lengend Snippet: Anticancer effect of Daemonorops draco Blume (DD) and reactivation of c-Jun and CCAAT/enhancer-binding protein-mediated apoptosis in acute myeloid leukemia cells. DD elevated the expression of miR-216b in (A) U937 and (B) THP-1 cells. AML cells were transfected with miR-216b inhibitor. (C–H) A total of 1 × 10 5 cells/ml were seeded into 6-well plates and allowed to reach approximately 50% density of transfection. The cells were transfected with miR-216b inhibitor for 48 h and exposed to the indicated doses of DD (i.e., 15 and 30 μg/ml) for 24 h. Following transfection for 48 h, miRNA was isolated and adopted to quantitative analysis of miRNA expression level or cell viability (E, F) , and Western blotting (G, H) was performed. Values represent the means of three experiments. Means ± standard deviation; * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with untreated control group. # p < 0.05, and ### p < 0.001 between the two groups.

Article Snippet: The AML cell lines THP-1 and U937 were purchased from Korean Cell Line Bank (Seoul, South Korea).

Techniques: Binding Assay, Expressing, Transfection, Isolation, Western Blot, Standard Deviation