沃度保兒謨ニ因セル濕疹

Evaluation of CNA-IC50 correlations by known cancer driver mutations. Distributions of PCCs for cancer cell lines with mutated or wild-type proto-oncogenes or tumor suppressors as depicted in each figure. The rank position of negatively correlated drugs (p < 0.10) is shown. (EPS 975 kb

A screen identifies PDMP as a drug that can synergistically inhibit the growth of human leukemia cells when combined with ABT-263.

<p>(<b>A</b>) Three human leukemia cell lines were screened against the LOPAC1280 compound library with and without the IC30 of ABT-737. A synergy score was calculated for each compound in the library and the scores of each compound are plotted as a component of each cell line. A lower score indicates a higher level of synergy. U937 cells are plotted on the x-axis, RPMI8226 cells are plotted on the y-axis and HL60 cells are plotted on the z-axis. The point on the graph representing PDMP is indicated. (<b>B</b>) Dose response curves of U937 cells were determined by treating the cells with either increasing doses of PDMP (350 nM to 45 µM) plus vehicle or increasing doses of PDMP (350 nM to 45 µM) and a constant dose of ABT-263 (2 µM). Inset values are the calculated IC50 from each curve. (<b>C</b>) Dose response curves of U937 cells were determined by treating the cells with either increasing doses of ABT-263 (8 nM to 18 µM) plus vehicle or increasing doses of ABT-263 (8 nM to 18 µM) and a constant dose of PDMP (45 µM). Arrows in (<b>B</b>) and (<b>C</b>) represent the equivalent doses of the respective drugs (2 µM ABT-263, 45 µM PDMP) and isobologram analysis indicated that the combination of the two drugs was synergistic with CI = <0.1. Inset values are the calculated IC50 from each curve. (<b>D</b>) Cells were treated with ABT-263 (2 µM), PDMP (45 µM) or the combination of drugs for 2, 4, or 8 hours and western blots for cleaved CASP3 were performed. (<b>E</b>) Cells were treated with either ABT-263 (2 µM), PDMP (45 µM) or the combination of drugs and 24 hours post treatment cells were stained with anti-AnnexinV antibody and 7AAD to determine the percent of cells undergoing apoptosis.</p

Treatment of U937 cells with PDMP, but not AMP-DNM causes accumuliation of ceramide and sphingosine and a decrease in glucosylceramide.

<p>(<b>A</b>) U937 cells were treated with ABT-263 (2 µM), PDMP (45 µM) or the combination of both drugs from two hours. Cells were harvested, lipids were extracted and the amounts of ceramide (Cer), hexosyceramine (HexCer) and lactosylceramide (LacCer) were quantitated. (<b>B</b>) U937 cells were treated with ABT-263 (2 µM), PDMP (45 µM) or the combination of both drugs from two hours. Cells were harvested, lipids were extracted and the amounts of sphingosine (Sph) and spingosine-1 phosphate (S1P) were quantitated. (<b>C</b>) U937 cells were treated with ABT-263 (2 µM), AMP-DNM (45 µM) or the combination of both drugs from two hours. Cells were harvested, lipids were extracted and the amounts of ceramide (Cer), hexosyceramine (HexCer) and lactosylceramide (LacCer) were quantitated. (<b>D</b>) U937 cells were treated with ABT-263 (2 µM), AMP-DNM (45 µM) or the combination of both drugs from two hours. Cells were harvested, lipids were extracted and the amounts of sphingosine (Sph) and spingosine-1 phosphate (S1P) were quantitated.</p

Increased ceramide and sphingosine are important for synergy with ABT-263.

<p>(<b>A</b>) Summary of the lipids that are altered following treatment of U937 cells with various inhibitors. (<b>B</b>) IC50 values of ABT-263 for individual human leukemia cell lines. Values were calculated in prism as the result of dose response curves determined by alamar blue assay 48 hours after ABT-263 treatment. (<b>C</b>) Total levels of basal ceramide (Cer) and sphingosine-1-phosphate (S1P) in four different cell lines as determined by HPLC-MS/MS. Data are normalized to the levels of lipids in U937. (<b>D</b>) Model depicting how different inhibitors affect sphingolipid metabolism.</p

Treatment of cells with AMP-deoxynojirimycin does not synergize with ABT-263.

<p>(<b>A</b>) and (<b>C</b>) Dose response curves of U937 cells and K562 were determined by treating the cells with either increasing doses of AMP (350 nM to 45 µM) plus vehicle or increasing doses of AMP (350 nM to 45 µM) and a constant dose of ABT-263 (2 µM or 60 nM, respectively). (<b>B</b>) and (<b>D</b>) Dose response curves of U937 cells and K562 cells were determined by treating the cells with either increasing doses of ABT-263 (8 nM to 18 µM for U937, 2.2 nM to 5 µM for K562) plus vehicle or increasing doses of ABT-263 (8 nM to 18 µM for U937, 2.2 nM to 5 µM for K562) and a constant dose of AMP (45 µM). Inset values are the calculated IC50 from each curve. (<b>E</b>) U937 cells were treated with ABT-263 (2 µM), PDMP (45 µM) or the combination of drugs for 8 hours and western blots for cleaved CASP3 were performed. (<b>F</b>) Cells were treated with either ABT-263 (60 nM), PDMP (45 µM) or the combination of drugs and 24 hours post treatment cells were stained with anti-AnnexinV antibody and 7AAD to determine the number of cells that were undergoing apoptosis or were already dead.</p

Validation of synergy in K562 cells, a line not used in the screen.

<p>(<b>A</b>) Dose response curves of K562 cells were determined by treating the cells with either increasing doses of PDMP (350 nM to 45 µM) plus vehicle or increasing doses of PDMP (350 nM to 45 µM) and a constant dose of ABT-263 (60 nM). Inset values are the calculated IC50 from each curve. (<b>B</b>) Dose response curves of K562 cells were determined by treating the cells with either increasing doses of ABT-263 (2.2 nM to 5 uM) plus vehicle or increasing doses of ABT-263 (2.2 nM to 5 µM) and a constant dose of PDMP (45 µM). Arrows in (A) and (B) represent equivalent doses of the respective drugs (60 nM ABT-263, 45 µM PDMP) and isobologram analysis indicated that the combination of the two drugs was synergistic with CI = <0.1. Inset values are the calculated IC50 from each curve. (<b>C</b>) Cells were treated with ABT-263 (60 nM), PDMP (45 µM) or the combination of drugs for 2, 4, or 8 hours and western blots for cleaved CASP3 were performed. (<b>D</b>) Cells were treated with either ABT-263 (60 nM), PDMP (45 µM) or the combination of drugs and 24 hours post treatment cells were stained with anti-AnnexinV antibody and 7AAD to determine the number of cells that were undergoing apoptosis or were already dead.</p

Drug combination analysis.

<p>A and B) Dose response curves for SAHA and Epirubicin; C) Median-effect plot; D) Algebraic estimate of the combinational index (CI) for the combination of epirubicin with SAHA relative to the fraction of affected cells.</p

Drugs-gene targets correlation and network.

<p>A) Drug name and number of high correlated genes mapped from the human PHEO and the murine MTT microarray data set. B) Connected node network representation in which all the targets of each drug are connected to the targets of all other drugs in the network.</p

Top-ranked compounds (hits) with high confidence antiproliferative activity in MTT PHEO cells.

<p>Top-ranked compounds (hits) with high confidence antiproliferative activity in MTT PHEO cells.</p

Validation of primary screening.

<p><i>Panel A and B:</i> Dose response curves of the selected compound in MPC, MTT and PC12 PHEO cell lines. The Y-axis represents signal intensity as a percent of the maximal value. The X-axis represents the log 10 concentrations of the respective compound (inside square brackets).</p