Journal: eLife

Article Title: BRAF V600E induces reversible mitotic arrest in human melanocytes via microrna-mediated suppression of AURKB

doi: 10.7554/eLife.70385

Figure Lengend Snippet: ( A ) Representative image of 15 H&E-stained melanocytic nevus. Border colors indicate two consecutive magnifications. Arrowheads indicate bi- (red) or multi- (blue) nucleation. ( B ) Example images of skin specimen containing a melanocytic nevus (circled) and phase-contrast microscopy of melanocytes isolated from nevus portion. Scale bar=400 µm. ( C ) Images of MLANA (purple) and Hoechst (green) co-staining of melanocytic nevus derived melanocytes in culture. Zoomed images show representative 2 N, 4 N, and 8 N cells. ( D ) Representative images of QPI coupled with fluorescence microscopy to identify adjacent mCherry-expressing and mCherry-negative melanocytes. Individual cells were identified at time zero, tracked, and monitored for division for 96 hr. ( E ) Mean and individual matched data points for the percent of n=3 mCherry-negative or mCherry-positive cells identified at time zero that divided over 96 hr. P value from paired t-test. ( F ) Mean and standard deviation for 48 hr growth rates of indicated cells treated with Barasertib. P values from unpaired t-tests comparing the 30 nM samples of melanoma lines to primary melanocytes (n=3). ( G ) Mean and standard deviation for relative QPI-derived dry mass per 501MEL cell at initial time point (black) compared to 48 hr treatment with Barasertib (gray) (n = 3). P values from unpaired t-tests. QPI, quantitative phase imaging.

Article Snippet: For each experiment, 100,000 melanocytes, 60,000 501Mel cells, or 150,000 HCIMel019 cells were plated per well and media containing indicated concentrations of barasertib (AURKB inhibitor; AZD1152-HQPA | AZD2811, Selleckchem, A1147) was added.

Techniques: Staining, Microscopy, Isolation, Derivative Assay, Fluorescence, Expressing, Standard Deviation, Imaging

Journal: eLife

Article Title: BRAF V600E induces reversible mitotic arrest in human melanocytes via microrna-mediated suppression of AURKB

doi: 10.7554/eLife.70385

Figure Lengend Snippet:

Article Snippet: For each experiment, 100,000 melanocytes, 60,000 501Mel cells, or 150,000 HCIMel019 cells were plated per well and media containing indicated concentrations of barasertib (AURKB inhibitor; AZD1152-HQPA | AZD2811, Selleckchem, A1147) was added.

Techniques: Derivative Assay, Western Blot, Recombinant, Sequencing, Control

Journal: Oncotarget

Article Title: Discovery of BPR1K871, a quinazoline based, multi-kinase inhibitor for the treatment of AML and solid tumors: Rational design, synthesis, in vitro and in vivo evaluation

doi: 10.18632/oncotarget.13369

Figure Lengend Snippet: Anti-proliferative activity of BPR1K871 on a panel of in-house leukemia cell lines

Article Snippet: Many research groups have reported small molecule dual FLT3 and AURK inhibitors that could be beneficial for the treatment of AML; for example, barasertib developed by AstraZeneca and currently in phase II/III clinical trials [ ]; CCT241736 developed by the Cancer Research UK Cancer Therapeutics Unit and currently in the preclinical stage [ ]; an indolinone derivative from Chern et al. [ ]; and a pyrrolopyrimidine derivative from our laboratory [ ].

Techniques: Activity Assay

Journal: Oncotarget

Article Title: Discovery of BPR1K871, a quinazoline based, multi-kinase inhibitor for the treatment of AML and solid tumors: Rational design, synthesis, in vitro and in vivo evaluation

doi: 10.18632/oncotarget.13369

Figure Lengend Snippet: Anti-proliferative activity of BPR1K871 on a panel of in-house non-leukemia cancer cell lines

Article Snippet: Many research groups have reported small molecule dual FLT3 and AURK inhibitors that could be beneficial for the treatment of AML; for example, barasertib developed by AstraZeneca and currently in phase II/III clinical trials [ ]; CCT241736 developed by the Cancer Research UK Cancer Therapeutics Unit and currently in the preclinical stage [ ]; an indolinone derivative from Chern et al. [ ]; and a pyrrolopyrimidine derivative from our laboratory [ ].

Techniques: Activity Assay

Journal: bioRxiv

Article Title: An antibiotic that mediates immune destruction of senescent cancer cells

doi: 10.1101/2024.09.05.611241

Figure Lengend Snippet: (a). Senolytic indexes of OXPHOS and mitochondrial complex inhibitors, mitochondrial function inhibitors, ER stress modulatory drugs and calcium regulatory drugs are determined by dividing the IC50 value of the drugs in parental A549 cells by the IC50 value in alisertib-induced senescent A549 cells. (b). Cell viability of alisertib-, barasertib-, etoposide- and CFI-400945-induced senescent A549 cells treated with salinomycin was assessed by colony formation assay. Proliferating cells were taken as a control. The cell viability was quantified (right panel). n = 3 independent experiments. Error bars represent mean ± standard deviation. Statistical significance was calculated with Two-Way ANOVA but is not shown in the graph. (c). Bobble plot showing pathway enrichment of calcium channel activity, calcium channel complex, calcium signaling pathway, mitochondrial complex homeostasis, OXPHOS, PERK, IRE1a, ATF6, UPR response to calcium ion and cytosolic calcium regulation in alisertib-induced senescent A549 treated with 2.5 µM salinomycin for 48 hours compared to untreated cells. The size of the circle indicates the p-value. (d). Volcano plot showing differentially expressed proteins related to OXPHOS subunits (red) and Unfolded Protein Response and ER stress (green) in alisertib-induced senescent A549 cells treated with 2.5 µM salinomycin for 48 hours compared to untreated cells. Thresholds for significance are -1.2 and 1.2 for abundance difference and 1.3 for -log 10 P. (e). Flow cytometry was used to determine the intensity of Ca 2+ dye Rhod-2 in parental and alisertib induced senescent A549 cells treated with 2.5 µM salinomycin for 4 hours. Untreated cells were taken as a control. n = 3 independent experiments. Error bars represent the mean ± standard deviation. Two-tailed unpaired student’s t-test was performed to calculate statistical significance. (f). Oxygen consumption rate (OCR) in proliferating A549 treated after 24 hours of 2.5 µM salinomycin treatment was measured using Seahorse assay. Untreated cells were taken as a control. Basal OCR and maximal respiratory capacity were quantified. For both n = 3 independent experiments, error bars represent mean ± standard deviation. Two-tailed unpaired student’s t-test was performed to calculate statistical significance. (g). Flow cytometry determined the percentage of ROS-positive cells in proliferating and alisertib-induced senescent A549 cells after 24 hours of 2.5 µM salinomycin treatment. Untreated proliferating cells were taken as a control. n = 3 independent experiments. Error bars represent the mean ± standard deviation. One-way ANOVA was performed to calculate statistical significance.

Article Snippet: Alisertib (#201931), barasertib (#200420), etoposide (#100330), PF-06873600 (#206997), Q-VD-OPh, (#526845), Z-VAD-FMK (#471029), necrosulfonamide (#525389), MCC950 (#522637) and 3-methyladenine (#406971) were purchased from MedKoo Biosciences Inc. CFI-400945 (#S7552) was purchased from Selleck Chemicals.

Techniques: Colony Assay, Control, Standard Deviation, Activity Assay, Flow Cytometry, Two Tailed Test