Consequently, ABT promotes cell death following treatment with agents that induce proapoptotic signals. ABT also exhibits significant singleagent activity against myeloma, leukemia, and small cell lung cancer cells in vitro is observed in many other cancer cells types. We have investigated the activity of ABT in ovarian cancer cells.All cell lines were grown in RPMI supplemented with FCS with the exception of OVCAR cells, which were grown in RPMI with FCS, and IGROV, which were cultured in DMEM with FCS.The proportion of the cells stained with the antibodies was determined by counting between and cells per ascites sample.ABT or its inactive enantiomer ABTE was prepared as a mmolL solution in DMSO.After h, the culture medium was removed and the cells fixed in TCA on ice for min.The following day, the cells were treated with drug or solvent and the volume adjusted to AL with FCSRPMI.After h, nucleosomes were quantified according to the manufacturers instructions.Detached cells were collected and the adherent cells collected by trypsinization.Both populations of cells were recombined, washed with PBS, and the cells lysed as described. Tumors were established from tissuecultured cells released from plastic flasks by a short exposure to trypsin, washed, and resuspended in PBS.Dosing Pregnenolone commenced when tumors were well established. Tumors were measured across two perpendicular diameters and volumes calculated from the following formula: V pd d as previously described. These include the ovarian cancer cell lines in the NCI panel and possess a range of sensitivities to carboplatin. These cells were rendered resistant to cisplatin by prolonged exposure of A cells to cisplatin in vitro. ABT inhibited the growthsurvival of each of the cell lines with comparable potencies between the cell lines.ABTE also inhibited cell growthsurvival, but to fold less potently than ABT. ABT sensitizes ovarian cancer cells to carboplatin.For initial combination experiments, ovarian cancer cell lines were treated simultaneously with solvent or ABT at a concentration selected to inhibit growthsurvival by, after which the potency of either paclitaxel or carboplatin were then determined. There was no increase in sensitivity to paclitaxel in any of the cell lines treated with ABT. In contrast, in most of the cell lines, ABT increased the sensitivity to carboplatin, and most prominently in IGROV, OVCAR, and OVCAR cells in which an ffold increase in sensitivity was observed. To confirm this, and in many of the subsequent experiments, IGROV cells were used because they showed the greatest increase in sensitivity to carboplatin when treated with ABT. IGROV cells were treated with a high concentration of carboplatin and examined by photomicroscopy.Inclusion of ABT decreased the time at which cell death became evident; after hof treatment with ABT and carboplatin, almost all of the cells were detached or displayed plasma membrane Diethylstilbestrol blebbing, whereas cells treated with carboplatin appeared to be morphologically unchanged. However, after h, almost all the cells treated with carboplatin alone were also detached.To quantify this, cells were treated with carboplatin and the attached cells were collected by trypsinization and counted.Inclusion of ABT significantly decreased the number of attached cells measured at both and h, consistent with the increase in drug sensitivity previously observed.

Leave a Reply