human leukemic cell lines hl60  (ATCC)

Journal: Clinical Epigenetics

Article Title: H19 overexpression promotes leukemogenesis and predicts unfavorable prognosis in acute myeloid leukemia

doi: 10.1186/s13148-018-0486-z

Figure Lengend Snippet: The biological role of H19 on leukemic cell line HL60. a The underlying role of H19 in leukemogenesis determined by Coremine Medical online database ( http://www.coremine.com/medical/ ). b H19 expression in eight common leukemic cell lines. The dotted line indicated the cutoff value to define H19 overexpression. c H19 expression mediated by siRNA-based H19 knockdown. H19 expression was significantly downregulated in siRNA-based H19 knockdown (siH19-1 and siH19-2) and control (siNC) groups. d The effect of H19 knockdown on cell proliferation. The si H19 groups (si H19 -1 and si H19 -2) showed significantly lower proliferation capacity than the siNC group at 48 and 72 h. e The effect of H19 knockdown on cell apoptosis. The si H19 groups (siH19-1 and siH19-2) showed significantly higher apoptosis rate than the siNC group at 48 h. f – h Flow-type apoptosis figures for siNC, si H19 -1, and si H19 -2, respectively. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: Human leukemic cell line HL60 (American Type Culture Collection, Manassas, VA, USA) was cultured in RPMI 1640 medium (BOSTER, Wuhan, China) containing 10% fetal calf serum (ExCell Bio, Shanghai, China) and grown at 37 °C in 5% CO 2 humidified atmosphere.

Techniques: Expressing, Over Expression, Knockdown, Control

Journal: Clinical Epigenetics

Article Title: H19 overexpression promotes leukemogenesis and predicts unfavorable prognosis in acute myeloid leukemia

doi: 10.1186/s13148-018-0486-z

Figure Lengend Snippet: Relationship between H19 and ID2 in AML. a ID2 mRNA expression in after siRNA-based H19 knockdown in HL60 cell line. ID2 mRNA expression was significantly downregulated after siRNA-based H19 knockdown (siH19-1 and siH19-2) and control (siNC) groups. ** P < 0.01, *** P < 0.001. b Correlation between H19 and ID2 expressions in AML patients. Correlation analysis was performed by Spearman test

Article Snippet: Human leukemic cell line HL60 (American Type Culture Collection, Manassas, VA, USA) was cultured in RPMI 1640 medium (BOSTER, Wuhan, China) containing 10% fetal calf serum (ExCell Bio, Shanghai, China) and grown at 37 °C in 5% CO 2 humidified atmosphere.

Techniques: Expressing, Knockdown, Control

Journal: Leukemia

Article Title: Heterogeneous sensitivity of human Acute Myeloid Leukemia to ß-catenin down-modulation

doi: 10.1038/leu.2011.17

Figure Lengend Snippet: (A) Silencing of ß-catenin (left panel) at the mRNA level in HL60 sorted GFP + cells at D4 post lentiviral transduction. 293T cells were used as an untransduced positive control; (mid panel) protein level obtained by flow cytometry in HL60 GFP + cells. ß-catenin fluorescence intensity is shown for the isotype control (filled line), sh ß-catenin GFP + (grey bold line) and control GFP + (black line); (right panel) protein level obtained by Western Blot. Scramble construct was used as control, sh22 served as sh ß-catenin. (B) HL60 GFP + proliferation kinetics at day 4, 6 and 8 (left panel). sh ß-catenin was sh47 versus respective scramble control. * p <0.05; (right panel) Cell cycle analysis at day 8. (C) Annexin V + fraction in GFP + HL60 cells as determined by flow cytometry on the same proliferation kinetics as in panel B. (D) Effect of ß-catenin knockdown on morphology, number of colonies ( p =0.01) in CFU assay. Original magnification ×90 and (E) Myeloid differentiation assay. Cells were sorted for GFP + for control or sh ß-catenin and exposed to 1μM ATRA for 4 days. Control: scramble construct; sh ß-catenin: sh22. Means are shown ± standard error of mean (SEM). * p <0.05.

Article Snippet: The HL60 human leukemic cell line was obtained from American Type Culture Collection, USA.

Techniques: Transduction, Positive Control, Flow Cytometry, Fluorescence, Control, Western Blot, Construct, Cell Cycle Assay, Knockdown, Colony-forming Unit Assay, Differentiation Assay

Journal: Leukemia

Article Title: Heterogeneous sensitivity of human Acute Myeloid Leukemia to ß-catenin down-modulation

doi: 10.1038/leu.2011.17

Figure Lengend Snippet: NOD/SCID mice were injected intra tibia with HL60 cell line after double transduction either with scramble control and luciferase (Luc) reporter constructs or with sh ß-catenin and luciferase reporter constructs. (A) Representative ventral view of BLI kinetics in a mouse injected with control GFP + Luc + transduced HL60 cells (left panel). Ventral and dorsal views of the same mouse at week 4 are shown on the right panel. Optical signal was expressed as photon flux (photons per second (p/s)) per cm 2 body surface analyzed. (B) Relative BLI signal fold change versus day 0 (H24). The values are the mean of the sum of measured signal from both ventral and dorsal positions at each time point (weeks 1 to 4), divided by the initial basal value at 24 hours. Control: HL60 scramble GFP + Luc + mice (n=6). sh ß-catenin: HL60 sh22 GFP + Luc + mice (n=6). * p <0.05. (C) Effect of ß-catenin silencing on week 4 engraftment of HL60 in NOD/SCID mice ( p =0.002). The engraftment was defined by the presence of CD45 + CD33 + CD19 − population. * p <0.05.

Article Snippet: The HL60 human leukemic cell line was obtained from American Type Culture Collection, USA.

Techniques: Injection, Transduction, Control, Luciferase, Construct

Journal: Journal of Hematology & Oncology

Article Title: Developing aptamer probes for acute myelogenous leukemia detection and surface protein biomarker discovery

doi: 10.1186/1756-8722-7-5

Figure Lengend Snippet: Aptamer recognition of cultured NB4 and HL60 leukemic cells. (a) . Comparison of aptamer recognition of cultured NB4 and HL60 leukemic cells. Individually synthesized biotin-labelled aptamers and PE-streptavidin were analyzed with flow cytometry in order to compare their ability to recognize NB4 and HL60 cells. Single-stranded library DNA was used as a negative control. The binding of selected aptamers with cells is illustrated as the following: negative control (black); JH6 (green); JH19 (blue); K19 (red). The final concentration of these aptamers in binding buffer was 150 nM. (b) . Determination of the aptamer binding affinities to NB4 cells. The biotin-labeled aptamers and PE-labeled streptavidin were used for the binding assays. The background binding was measured by using unselected single-stranded library DNA. The fluorescence intensity geometric means of bound aptamers was determined by flow cytometry. The equilibrium dissociation constants (Kd) of the fluorescent ligands were obtained by fitting the dependence of specific binding fluorescence intensity on the concentration of the ligands to the Equation Y = Bmax*X/(Kd + X) using the GraphPad Software (San Diego, CA, USA) as described in previous studies .

Article Snippet: NB4 and HL60 human leukemic cell lines were obtained from ATCC (American Type Culture Collection, Manassas, Virginia) and were cultured in RPMI 1640 medium (Thermo Scientific HyClone, South Logan, Utah) supplemented with 10% fetal bovine serum (FBS) (heat inactivated, Thermo Scientific HyClone, South Logan, Utah), and antibiotics (100 units/ml penicillin-Streptomycin from Fisher BioReagents, Fairlawn, NJ).

Techniques: Cell Culture, Comparison, Synthesized, Flow Cytometry, Negative Control, Binding Assay, Concentration Assay, Labeling, Fluorescence, Software