human leukemia hl 60 aml  (ATCC)

Journal: EJHaem

Article Title: GSK3 inhibitor enhances gemtuzumab ozogamicin‐induced apoptosis in primary human leukemia cells by overcoming multiple mechanisms of resistance

doi: 10.1002/jha2.600

Figure Lengend Snippet: Cytotoxic effect of gemtuzumab ozogamicin (GO) was enhanced by GSK3α/β inhibitors treatment in acute myeloid leukemia (AML) cell lines. For all experiments, specific apoptosis was determined after 48 h of incubation. The resulting data were expressed as means ± standard deviation (SD) of three independent experiments. (A) U937 and MARIMO cells were treated with the indicated concentrations of CHIR99021 (CHIR). Apoptosis was determined by staining with Annexin V and Propidium Iodide (PI), followed by flow cytometry. (B) U937 and MARIMO cells were treated with the indicated concentrations of CHIR and then analyzed for accumulation of β‐catenin by western blotting. (C) Cells were treated with 2.5 μg/ml, 0.5 μg/ml (for THP‐1), or 0.25 μg/ml (for NB4) of GO alone, or in combination with CHIR. *, **, and *** indicate p < 0.05, p < 0.01, and p < 0.001, respectively. (D) Cells were treated with GO, CHIR, or GO+CHIR. Whole cell lysates were analyzed by western blotting for poly(ADP‐ribose) polymerase (PARP). β‐actin was used as a loading control. (E) MARIMO, KO52, and U937 cells were treated with GO, AZD2858 (A2848), or GO+A2858. The statistical significance of differences observed between GO and GO+A2858 were determined using Student's t ‐test. *and *** indicate p < 0.05 and p < 0.001, respectively. (F) MARIMO, KO52, and U937 cells were treated with GO, AZD1080 (A1080), or GO+A1080.The statistical significance of differences observed between GO and GO+A1080 was determined using Student's t ‐test. *, ** and *** indicate p < 0.05, p < 0.01, and p < 0.001, respectively.

Article Snippet: Human AML cell lines human promyelocyte leukemia cell line (HL‐60) and KO52 (JCRB Cell Bank), U937 American Type Culture Collection (ATCC), human monocytic leukemia cell line (THP‐1) (ECACC), MARIMO (kind gift from Dr. Yosuke Minami at the Department of Hematology/Oncology, Kobe University, Kobe, Japan), and NB4 (kind gift from Dr. Hideki Nakakuma at the Department of Hematology/Oncology, Wakayama Medical University, Wakayama, Japan) were cultured as previously described [ , ].

Techniques: Incubation, Standard Deviation, Staining, Flow Cytometry, Western Blot, Control

Journal: EJHaem

Article Title: GSK3 inhibitor enhances gemtuzumab ozogamicin‐induced apoptosis in primary human leukemia cells by overcoming multiple mechanisms of resistance

doi: 10.1002/jha2.600

Figure Lengend Snippet: GSK3α/β inhibitor increases CD33 expression on acute myeloid leukemia (AML) cell lines. (A) Flow cytometric analysis of CD33 expression after 48 h at different concentrations of CHIR99021. (B) The bar graph shows CD33 expression at different drug concentrations. The heights of the bars are CD33 mean fluorescence intensity (MFI) from CHIR‐treated cells normalized to CD33 MFI of untreated cells. Data are expressed as means ± SD of three independent experiments. *, ** and *** indicate p < 0.05, p < 0.01, and p < 0.001, respectively. (C) U937 cells were treated with CHIR 5 μM and analyzed CD33 expression by confocal microscopy.

Article Snippet: Human AML cell lines human promyelocyte leukemia cell line (HL‐60) and KO52 (JCRB Cell Bank), U937 American Type Culture Collection (ATCC), human monocytic leukemia cell line (THP‐1) (ECACC), MARIMO (kind gift from Dr. Yosuke Minami at the Department of Hematology/Oncology, Kobe University, Kobe, Japan), and NB4 (kind gift from Dr. Hideki Nakakuma at the Department of Hematology/Oncology, Wakayama Medical University, Wakayama, Japan) were cultured as previously described [ , ].

Techniques: Expressing, Fluorescence, Confocal Microscopy

Journal: Heliyon

Article Title: Combination of coumarin and doxorubicin induces drug-resistant acute myeloid leukemia cell death

doi: 10.1016/j.heliyon.2021.e06255

Figure Lengend Snippet: Western bloting of Cas-3, P53, PARP and Actin in HL60/ADR after co-treatment of 0.1 μg/ml of doxirubcin with (100, 250, 500, and 1000 μg/ml) coumarin) and (B-E) a quantification of western blot data using Image J softrware.

Article Snippet: Both cell lines used (Acute human myeloid leukemia (AML) HL60 and HL60/ADR) were purchased from ATCC (American Tissue Culture Collection).

Techniques: Western Blot

Journal: Heliyon

Article Title: Combination of coumarin and doxorubicin induces drug-resistant acute myeloid leukemia cell death

doi: 10.1016/j.heliyon.2021.e06255

Figure Lengend Snippet: (A) cytotoxic effect of coumarin and doxrubcin treatment on HL60 cells using trypane blue exclusion dye. Apoptotic and necrotic cell death were assessed in HL60 using Annexin V and PI staining and analyzed using flow cytometer in ( B) negative control, (C) treated with 0.1 μg/ml of doxirubcin as a positive control, and (D -G) treated with (100, 250, 500, and 1000 μg/ml) coumarin.

Article Snippet: Both cell lines used (Acute human myeloid leukemia (AML) HL60 and HL60/ADR) were purchased from ATCC (American Tissue Culture Collection).

Techniques: Staining, Flow Cytometry, Negative Control, Positive Control

Journal: Heliyon

Article Title: Combination of coumarin and doxorubicin induces drug-resistant acute myeloid leukemia cell death

doi: 10.1016/j.heliyon.2021.e06255

Figure Lengend Snippet: Cell viability was assessed in HL60/ADR cells using trypane blue exclusion dye; (A) treated with different coumarin concentrations and (B) co-treated with 100 ng/ml doxrubcin with different coumarin concentrations.

Article Snippet: Both cell lines used (Acute human myeloid leukemia (AML) HL60 and HL60/ADR) were purchased from ATCC (American Tissue Culture Collection).

Techniques:

Journal: Heliyon

Article Title: Combination of coumarin and doxorubicin induces drug-resistant acute myeloid leukemia cell death

doi: 10.1016/j.heliyon.2021.e06255

Figure Lengend Snippet: Apoptotic and necrotic cell death were assessed in HL60/ADR cells using Annexin V and PI staining and analyzed using flow cytometer in ( B) (negative control), (C-F) HL60/ADR cells treated with (100, 250, 500, and 1000 μg/ml) coumarin , and (G-J) combination of 0.1 μg/ml of doxirubcin with (100, 250, 500, and 1000 μg/ml) coumarin and (K) quantification of Flowcytometry results.

Article Snippet: Both cell lines used (Acute human myeloid leukemia (AML) HL60 and HL60/ADR) were purchased from ATCC (American Tissue Culture Collection).

Techniques: Staining, Flow Cytometry, Negative Control

Journal: Heliyon

Article Title: Combination of coumarin and doxorubicin induces drug-resistant acute myeloid leukemia cell death

doi: 10.1016/j.heliyon.2021.e06255

Figure Lengend Snippet: Confocal imaging of ROS in HL60/ADR after treatment with; (A) negative control, (B) 0.1 μg/ml of doxirubcin, (C) 500 μM Pyocyanin, (D-G) treated with (100, 250, 500, and 1000 μg/ml) coumarin, and (H-K) combination of 0.1 μg/ml of doxirubcin with (100, 250, 500, and 1000 μg/ml) coumarin and (L) quantification of the intensity of fluorescence using imge J software.

Article Snippet: Both cell lines used (Acute human myeloid leukemia (AML) HL60 and HL60/ADR) were purchased from ATCC (American Tissue Culture Collection).

Techniques: Imaging, Negative Control, Fluorescence, Software