McEleny Kevin, Coffey Ronan, Morrissey Colm, Williamson Kate, Zangemeister-Wittke Uwe, Fitzpatrick John M, Watson R William G
Department of Surgery, Mater Misericordiae University Hospital and Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Ireland.
Prostate. 2004 Jun 1;59(4):419-25. doi: 10.1002/pros.10371.
The inhibitors of apoptosis (IAP) proteins are a family of structurally homologous caspase inhibitors. We synthesized an antisense oligonucleotide (AO) to target a region within the BIR domain of cIAP-1 and examined its ability to facilitate apoptosis in prostate cancer cells.
We transfected the IAP AO into PC3 and DU145 cells and determined alterations in IAP expression using Western blotting. Apoptosis and viability were assessed using propidium iodide (PI) DNA incorporation with flow cytometry. Pacitaxel, caffeic acid phenethyl ester (CAPE), Fas antibody, and TNFalpha were used as 'second hit' agents in association with the AO.
Western blotting showed a down-regulation in cIAP-1 expression and higher levels of spontaneous apoptosis in both cell types with no alteration in overall cell viability. AO sensitized PC3 cells, to Fas antibody and TNFalpha-mediated apoptosis, but not to apoptosis mediated by paclitaxel or CAPE.
cIAP-1 down-regulation increased spontaneous apoptosis in prostate cancer cells and sensitized PC3 cells to receptor-mediated apoptosis.
凋亡抑制蛋白（IAP）家族是一类结构同源的半胱天冬酶抑制剂。我们合成了一种反义寡核苷酸（AO），靶向cIAP-1的BIR结构域内的一个区域，并检测其促进前列腺癌细胞凋亡的能力。
我们将IAP AO转染到PC3和DU145细胞中，并用蛋白质免疫印迹法检测IAP表达的变化。使用碘化丙啶（PI）掺入法结合流式细胞术评估细胞凋亡和活力。紫杉醇、咖啡酸苯乙酯（CAPE）、Fas抗体和肿瘤坏死因子α（TNFα）与AO联合用作“二次打击”试剂。
蛋白质免疫印迹法显示，两种细胞类型中cIAP-1表达均下调，自发凋亡水平更高，总体细胞活力无变化。AO使PC3细胞对Fas抗体和TNFα介导的凋亡敏感，但对紫杉醇或CAPE介导的凋亡不敏感。
cIAP-1下调增加了前列腺癌细胞的自发凋亡，并使PC3细胞对受体介导的凋亡敏感。
