Splicing reprogramming of TRAIL/DISC-components sensitizes lung cancer cells to TRAIL-mediated apoptosis

摘要

a Cell viability was evaluated using MTT assays in A549 and Calu-1 human NSCLC cell lines and normal nontransformed primary human lung fibroblasts (LF) pretreated with 10, 25, or 50 μM apigenin or DMSO (diluent control, noted as 0) or for 18 h followed by treatment with TRAIL (25, 50, or 100 ng/ml) for additional 6 h. Cell viability was also evaluated in A549 cells pretreated with 10, 25, or 50 μM genistein or diluent for 18 h followed by TRAIL treatment for additional 6 h. b Cell viability was evaluated in PHLECs isolated from paired tumor tissue biopsies (PHLEC-ACCs) and nontumor tissue biopsies (nontumor PHLECs) from three subjects (P1–P3) and pretreated with 10, 25, or 50 μM apigenin or diluent for 18 h followed by 25, 50, or 100 ng/ml TRAIL for an additional 6 h. c–f The percentage of cells stained with AnnexinV/7-AAD (c) or active caspase-3 (e), which are markers of apoptosis, was evaluated in PHLEC-ACCs isolated from P1 to P3. Cells were pretreated with 50 μM apigenin or DMSO (indicated as -) for 18 h followed by 100 ng/ml TRAIL for 6 h, and then quantification was performed with flow cytometry. A representative flow cytometry plot of cells isolated from P1 stained with AnnexinV/7-AAD (d) or active caspase-3 and 7-AAD (f). All data are presented as the mean ± SEM (n = 3; **P < 0.01, and ***P < 0.001; and analyses were performed with two-way ANOVA).