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目的:研究TRAIL诱导激素非依赖前列腺癌细胞株PC-3M过程中核因子kappaB(NF-κB)的活化和失活现象。方法:当不同浓度的TRAIL和LPS作用于细胞后,我们通过细胞免疫组化染色和凝胶电泳迁移试验(EMSA)来检测NF-κB核转位的情况。并通过RT-PCR的方法粗略评定二硫代氨基甲酸吡咯烷(PDTC)对抑制蛋白IκB的影响。结果:EMSA和免疫组化分析显示PC-3M细胞中NF-κB的核转位可被TRAIL或LPS明显地激活。以PDTC预处理可以上调抑制蛋白IκB的表达,阻断NF-κB的核转位。结论:TRAIL的作用于激素非依赖前列腺癌细胞时主要的负作用在于其可以显著地刺激NF-κB的活化。另一方面,在PC-3M细胞凋亡过程中NF-κB的核转位可以被PDTC有力地抑制,同时IκB的表达升高和降解减少(PDTC引起的)是抑制NF-κB活化的潜在因素,为我们提出了增强TRAIL疗效的可能性方案。
AIM: To investigate the activation and inactivation of nuclear factor kappaB (NF-κB) during the TRAIL-induced hormone-independent prostate cancer cell line PC-3M. Methods: After different concentrations of TRAIL and LPS were applied to cells, we detected the nuclear translocation of NF-κB by immunohistochemical staining and electrophoretic mobility shift assay (EMSA). The effect of pyrrolidine dithiocarbamate (PDTC) on the inhibitory protein IκB was roughly evaluated by RT-PCR. Results: EMSA and immunohistochemical analysis showed that nuclear translocation of NF-κB in PC-3M cells was significantly activated by TRAIL or LPS. Pretreatment with PDTC upregulated the expression of IκB, and blocked the nuclear translocation of NF-κB. Conclusion: The main negative effect of TRAIL on hormone-independent prostate cancer cells is that it can significantly stimulate the activation of NF-κB. On the other hand, nuclear translocation of NF-κB during PC-3M cell apoptosis can be strongly inhibited by PDTC while increased expression of IκB and decreased degradation (caused by PDTC) are potential factors that inhibit NF-κB activation , Proposed for us the possibility of enhancing the efficacy of TRAIL program.