Journal: Bioengineered

Article Title: Circular RNA circ_0004277 Inhibits Acute Myeloid Leukemia Progression Through MicroRNA-134-5p / Single stranded DNA binding protein 2

doi: 10.1080/21655979.2022.2059609

Figure Lengend Snippet: Circ_0004277 expression is remarkably up-regulated in AML. A. Heat map showed the differentially expressed circRNAs (bone marrow mononuclear cells of AML patients v.s. bone marrow mononuclear cells of healthy controls, GSE94591), and the filter criteria was P < 0.05,Log2(Change fold)> 2.5. B. The expression of circ_0004277 in bone marrow mononuclear cells of AML patients was significantly down-regulated compared with that of healthy controls (GSE94591). C. qRT-PCR showed that circ_0004277 expression was lower in the bone marrow samples of 37 AML patients than that in 37 healthy controls. D. qRT-PCR revealed that circ_0004277 expression in AML cells (KG-1a, THP-1, K562 and U937) was down-regulated compared with the human normal stromal cells (HS-5). E. qRT-PCR showed that circ_0004277 was resistant to RNase R. ** P < 0.01, and *** P < 0.001.

Article Snippet: Human bone marrow stromal cell line HS-5 and human AML cell lines (KG-1a, THP-1, K562 and U937) were procured from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Quantitative RT-PCR

Journal: Bioengineered

Article Title: Circular RNA circ_0004277 Inhibits Acute Myeloid Leukemia Progression Through MicroRNA-134-5p / Single stranded DNA binding protein 2

doi: 10.1080/21655979.2022.2059609

Figure Lengend Snippet: Circ_0004277 directly targets miR-134-5p. A. Bioinformatics was employed to predict the binding site between circ_0004277 and miR-134-5p. B. Dual-luciferase reporter gene assay assays demonstrated that miR-134-5p could negatively regulate the luciferase activity of circ_0004277-WT, rather than circ_0004277-MUT in U937 and KG-1a cells. C. RIP assay indicated that circ_0004277 and miR-134-5p were enriched in anti-Ago2 group, suggesting circ_0004277 and miR-134-5p were directly interacted. D. RNA pull-down assay showed that circ_0004277 and miR-134-5p could combine with each other. E. qRT-PCR indicated that miR-134-5p expression was decreased in the circ_0004277 overexpression plasmid group compared with the empty plasmid group. F. qRT-PCR indicated that miR-134-5p expression was increased in the bone marrow samples of 37 AML patients compared with that of 37 healthy donors. G. circ_0004277 expression in AML tissues was negatively correlated with miR-134-5p expression in bone marrow samples of the patients with AML. ** P < 0.01, and *** P < 0.001.

Article Snippet: Human bone marrow stromal cell line HS-5 and human AML cell lines (KG-1a, THP-1, K562 and U937) were procured from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Binding Assay, Luciferase, Reporter Gene Assay, Activity Assay, Pull Down Assay, Quantitative RT-PCR, Expressing, Over Expression, Plasmid Preparation

Journal: Bioengineered

Article Title: Circular RNA circ_0004277 Inhibits Acute Myeloid Leukemia Progression Through MicroRNA-134-5p / Single stranded DNA binding protein 2

doi: 10.1080/21655979.2022.2059609

Figure Lengend Snippet: MiR-134-5p inhibits the proliferation and aggressiveness of KG-1a and U937 cells. A. qRT-PCR showed that miR-134-5p expression was up-regulated in AML cell lines (KG-1a, THP-1, K562 and U937) compared with the human normal stromal cells (HS-5). B. qRT-PCR revealed that miR-134-5p expression was down-regulated in U937 and KG-1a cells transfected with miR-134-5p inhibitor. C. CCK-8 assay showed the viability of KG-1a and U937 cells was significantly decreased after the transfection of anti-miR-134-5p. D. Transwell assay indicated that the migration and invasion of KG-1a and U937 cells were decreased after the transfection of anti-miR-134-5p. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Human bone marrow stromal cell line HS-5 and human AML cell lines (KG-1a, THP-1, K562 and U937) were procured from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Quantitative RT-PCR, Expressing, Transfection, CCK-8 Assay, Transwell Assay, Migration

Journal: Bioengineered

Article Title: Circular RNA circ_0004277 Inhibits Acute Myeloid Leukemia Progression Through MicroRNA-134-5p / Single stranded DNA binding protein 2

doi: 10.1080/21655979.2022.2059609

Figure Lengend Snippet: Circ_0004277 modulates the biological behaviors of AML cells by sponging miR-134-5p. A. qRT-PCR showed that the miR-134-5p mimics have no effect on the expression of circ_0004277. B. qRT-PCR indicated that miR-134-5p mimics could up-regulate the expression of miR-134-5p. C. CCK-8 assay showed that miR-134-5p mimics could reverse the effect of circ_0004277 overexpression plasmid on the viability of U937 and KG-1a cells. D. Transwell assay showed that the migration and invasion of U937 and KG-1a cells transfected with circ_0004277 overexpression plasmid was reversed by miR-134-5p mimics. ** P < 0.01, *** P < 0.001.

Article Snippet: Human bone marrow stromal cell line HS-5 and human AML cell lines (KG-1a, THP-1, K562 and U937) were procured from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Quantitative RT-PCR, Expressing, CCK-8 Assay, Over Expression, Plasmid Preparation, Transwell Assay, Migration, Transfection

Journal: Frontiers in immunology

Article Title: NAD metabolism-related genes provide prognostic value and potential therapeutic insights for acute myeloid leukemia.

doi: 10.3389/fimmu.2024.1417398

Figure Lengend Snippet: FIGURE 6 Exploration of effective specific small-molecule inhibitors for the NADM8high patients. (A) Bar plot shows the distribution of chemotherapy response rate between the NADM8high and NADM8low patients in all the RJAML cohort and (B) the patients who received the “3 + 7”-based regimens as initial induction in the development cohort. (C) ROC curve indicates the prediction power of NADM8 for chemotherapy resistance in the univariate logistic regression model (AUC=0.67, P<0.001). (D) Illustrative diagram of ex vivo drug sensitivity data in AML patients from the BeatAML cohort. The freshly isolated mononuclear cells from patients were exposed to 155 small-molecule inhibitors at different concentrations and the drug sensitivity of these patient-derived cells was further determined. (E, F) Pearson’s correlation analysis identifies the potential drugs, GDC-0941 and AT-7519, that exert therapeutic effect in NADM8high patients. Significant inhibitors were determined using the threshold of p-value < 0.05. (G, H) GSEA shows the enrichment of PI3K-AKT-mTOR signaling and MAPK signaling pathway in NADM8high patients. (I) Heatmap depicts the expression level of the 8 NAD-related genes in AML cell lines. (J) The Pearson’s correlation plot shows that the NADM8 scores of the 9 AML cell lines are negatively associated with the IC50 score of GDC-0941 generated from in vitro drug sensitivity assays. (K) Boxplot showing IC50(mM) between samples of NADM8high and NADM8low patients against GDC-0941.

Article Snippet: A panel of AML cell lines (HL-60, U937, OCI-AML3, Kasumi-1, ME1, NB4, MOLM13, OCI-AML2, K562) were exposed to varying concentrations of GDC-0941 (Selleckchem, S2065) in 96-well plates at a density of 2 × 104 cells per well.

Techniques: Ex Vivo, Isolation, Derivative Assay, Expressing, Generated, In Vitro

Journal: Frontiers in immunology

Article Title: NAD metabolism-related genes provide prognostic value and potential therapeutic insights for acute myeloid leukemia.

doi: 10.3389/fimmu.2024.1417398

Figure Lengend Snippet: FIGURE 7 The functional exploration of SLC25A51 in AML. (A) Boxplot showing the average CRISPR scores across AML cell lines of the 8 genes in the NADM8 model. (B) Kaplan-Meier estimates of overall survival (OS) according to the expression level of SLC25A51 in the whole RJAML training cohort. (C) The overall survival status of the RJAML intermediate-risk patients between the two risk groups categorized by the median expression level of SLC25A51. (D, F) The knockdown efficiency of the SLC25A51 gene expression level in U937 cell line (D) and THP-1 cell line (F). (E, G) The proliferative capacity of U937 and THP-1 cells following stable knockdown of SLC25A51 as measured by CCK8 assays, with red and blue lines representing the knockdown and control vector group, respectively. (H, I) Knockdown of SLC25A51 could strongly induce apoptosis of U937 and THP-1 cells, as measured by annexin V/PI flow cytometric analyses. The histogram depicts the quantification of the total proportion of cells in the early (annexin V +/PI–) and late apoptotic (annexin V+/PI+) stages. All plots are representative of at least three independent experiments performed in duplicate and presented as the means ± standard deviation. (*p < 0.05, **p <0.01 and ***p < 0.001).

Article Snippet: A panel of AML cell lines (HL-60, U937, OCI-AML3, Kasumi-1, ME1, NB4, MOLM13, OCI-AML2, K562) were exposed to varying concentrations of GDC-0941 (Selleckchem, S2065) in 96-well plates at a density of 2 × 104 cells per well.

Techniques: Functional Assay, CRISPR, Expressing, Knockdown, Gene Expression, Control, Plasmid Preparation, Standard Deviation

Journal: Oncotarget

Article Title: Role of drug transporters in the sensitivity of acute myeloid leukemia to sorafenib

doi: 10.18632/oncotarget.25494

Figure Lengend Snippet: Characteristics of acute myeloid leukemia (AML) and K-562 cell lines

Article Snippet: Human cell lines HL-60, MOLM-13, HEL, OCI-AML2, derived from AML, and K-562 derived from chronic myelogenous leukemia in blast phase (CML-BP), were purchased from the DSMZ or ATCC cell line repositories and were grown in RPMI medium with 10% heat-inactivated fetal bovine serum (FBS), except OCI-AML2 cells in alpha-MEM medium and 20% FBS, supplemented with 100 U/ml penicillin and 100 μg/ml streptomycin, at 37°C and 5% CO 2 .

Techniques: