OSI-027 induces apoptosis in malignant lymphoid cell lines and clinical lymphoma samples. (A-C) Jeko and Mino cells were treated for 48 hours with the indicated concentrations of OSI-027 (B) or rapamycin (C) and assayed using annexin V and PI as illustrated in panel A. Values in panels B and C represent the percentage of total cells that were negative for staining with annexin V and PI. (D) Jurkat were treated for 48 hours with the indicated concentrations of OSI-027, lysed in citric acid under conditions where fragmented DNA is extracted, stained with propidium iodide (PI) and subjected to flow microfluorimetry. Results of this and additional assays are summarized in Figure 6A. (E-G) Jeko, SeAx and Jurkat cells were treated for 24-96 hours with the indicated concentrations of OSI-027 (E-F) or 10nM rapamycin (G) and assayed as illustrated in panel D. (H) After treatment with diluent or OSI-027 for 48 hours, lysates from Jeko cells, Mino cells, or MCL samples from 3 separate patients were subjected to SDS-PAGE and probed with antibody to PARP1. Arrow, previously described caspase-induced PARP cleavage fragment.48 Error bars in panels B, C, E, and F, mean ± SEM of 3 independent assays.
