molt16 cells  (Broad Institute Inc)

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Synergistic Drug Combinations with a CDK4/6 Inhibitor in T-cell Acute Lymphoblastic Leukemia

doi: 10.1158/1078-0432.CCR-15-2869

Figure Lengend Snippet: A) Ten T-ALL cell lines were grown in a range of LEE011 concentrations and viability evaluated at day 6 by an ATP-based assay as the percentage of viable cells relative to a DMSO control. Shown are the mean ± standard deviation (SD) of 4 replicates. B) Western immunoblotting showing expression of ICN1, RB, Cyclin D3, CDK4 and CDK6 in a panel of T-ALL cell lines. C) Table showing calculated IC50 values for LEE011 treatment from the dose-response curves in panel A. Also annotated is the ICN1 and RB1 status for each cell line. D) Western immunoblotting showing a decrease in phosphorylation of RB with increasing concentrations of LEE011 in ICN1 positive (MOLT4) and negative (MOLT16) cell lines. E) Cell cycle analysis in MOLT4 and MOLT16 cells treated with increasing concentrations of LEE011. F) Ratio of Annexin V positive cells to control with increasing concentrations of LEE011 treatment of MOLT4 and MOLT16 cells. Shown are the mean ± standard error of mean (SEM) of 2 separate experiments. G) Correlation of ratio of Annexin V positive cells (relative to control) versus ratio of Cell Titer-Glo (CTG) Luminescence (relative to control) of cells treated with increasing concentrations of LEE011 at the indicated doses. Numbers indicate LEE011 concentration in μM units.

Article Snippet: Cell culture, cell viability and apoptosis assays The human cell line MOLT16 was purchased from Leibniz-Institut DSMZ-German collection of microorganisms and cell cultures.

Techniques: ATP Assay, Control, Standard Deviation, Western Blot, Expressing, Phospho-proteomics, Cell Cycle Assay, Concentration Assay

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Synergistic Drug Combinations with a CDK4/6 Inhibitor in T-cell Acute Lymphoblastic Leukemia

doi: 10.1158/1078-0432.CCR-15-2869

Figure Lengend Snippet: Combination index analysis for the combinations of LEE011 with A) methotrexate, B) mercaptopurine, C) L-asparaginase and D) doxorubicin in MOLT4 and MOLT16 cells treated for 6 days in replicates of 4.

Article Snippet: Cell culture, cell viability and apoptosis assays The human cell line MOLT16 was purchased from Leibniz-Institut DSMZ-German collection of microorganisms and cell cultures.

Techniques:

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Synergistic Drug Combinations with a CDK4/6 Inhibitor in T-cell Acute Lymphoblastic Leukemia

doi: 10.1158/1078-0432.CCR-15-2869

Figure Lengend Snippet: A) A luciferized MOLT16 mouse model was treated with vehicle, dexamethasone (D) 15 mg/kg, LEE011 (L) 75 mg/kg, everolimus (E) 5mg/kg, or the combination of LEE011 with dexamethasone (L+D) or LEE011 with everolimus (L+E) after detectable disease was established. Spleen weight was analyzed in 3 mice per group after 5 days of treatment. Shown is the average spleen weight; error bars represent SD for 3 mice per group. B) Kaplan Meier curves showing overall survival of mice (n=10 per group) treated with vehicle, dexamethasone, LEE011 or the combination. * represents P-value <0.05 calculated using log-rank test. C) Kaplan Meier curves showing overall survival of mice (n=10 per group) treated with vehicle, everolimus, LEE011 or the combination. * shows P-value <0.05 calculated using log-rank test, *** shows P-value <0.001 calculated using log-rank test, **** shows P-value <0.0001 calculated using log-rank test. D) Mean fluorescence intensity (MFI) of p-4E-BP1 and E) pRB levels from peripheral blood cells measured by flow cytometry from mice treated for 5 days with drugs as indicated. Vehicle-treated mice are shown in black and drug-treated mice are shown in red. Each bar represents a single mouse. A luciferized MOLT4 mouse model was treated with vehicle, dexamethasone (D) 15 mg/kg, LEE011 (L) 75 mg/kg, or the combination of LEE011 with dexamethasone (L+D) after detectable disease was established. F) Spleen weight was analyzed from 3 mice per group after 5 days of treatment. Shown is the average spleen weight; error bars represent SD for 3 mice. * represents P-value <0.05 and ** represents P-value <0.005 calculated by paired t-test, n=3. G) Kaplan-Meier curves showing overall survival of mice (n=10 per group). P-value calculated using log-rank test. P-value <0.0001 between group treated with combination of LEE011 and dexamethasone and any individual treatment groups. H) Histology of mice treated with vehicle, LEE011, dexamethasone and combination of LEE011 and dexamethasone. Mice were treated for 5 days and tissue collected 3 hours after dosing. Shown are representative sections of bone marrow stained with p-RB S807/811. Images taken at 200× magnification.

Article Snippet: Cell culture, cell viability and apoptosis assays The human cell line MOLT16 was purchased from Leibniz-Institut DSMZ-German collection of microorganisms and cell cultures.

Techniques: Fluorescence, Flow Cytometry, Staining