Godlewski Michat Marek, Gajkowska Barbara, Lamparska-Przybysz Monika, Motyl Tomasz
Department of Physiological Sciences, Faculty of Veterinary Medicine, Warsaw Agricultural University, 02-776 Warsaw, Poland.
Anticancer Drugs. 2002 Nov;13(10):1017-29. doi: 10.1097/00001813-200211000-00006.
Cyclooxygenase (COX)-2 inhibitors that belong to non-steroid anti-inflammatory drug family have been shown to have an apoptosis-inducing effect on neoplastic cells. In the present study the effect of nimesulide (NIM), a specific COX-2 inhibitor, on apoptosis and interactions between BCL-2 family death promoters BAX and BID and BAX and VDAC-1 were examined in human colon adenocarcinoma COLO 205 cells. Laser scanning cytometry was applied for the measurement of expression and aggregation of apoptosis-related proteins and quantitative analysis of NIM-induced apoptosis. Double-staining immunoconfocal and immunoelectron microscopy were used for subcellular colocalization of examined proteins. NIM induced apoptosis of COLO 205 cells in a dose-dependent manner. This was accompanied by: (1) a decrease in intracellular prostaglandin (PG) E content; (2) subcellular redistribution and aggregation of BAX and BID on organellar membranes and within the nucleus; (3) colocalization of BAX with BID and BAX with VDAC-1 on organelles; and (4) survival of cells with the highest BCL-2 aggregation. A similar pattern of subcellular redistribution and colocalization of BAX with BID and BAX with VDAC-1 suggests that BAX (in association with BID) controls the function of VDAC-1 and its permeability for apoptogenic factors released from mitochondria of COLO 205 cells stimulated to apoptosis with NIM.
属于非甾体抗炎药家族的环氧化酶（COX）-2抑制剂已被证明对肿瘤细胞具有诱导凋亡的作用。在本研究中，检测了特异性COX-2抑制剂尼美舒利（NIM）对人结肠腺癌COLO 205细胞凋亡以及BCL-2家族促凋亡蛋白BAX与BID之间、BAX与VDAC-1之间相互作用的影响。应用激光扫描细胞术测量凋亡相关蛋白的表达和聚集，并对NIM诱导的凋亡进行定量分析。采用双染免疫共聚焦和免疫电子显微镜对检测蛋白进行亚细胞共定位。NIM以剂量依赖性方式诱导COLO 205细胞凋亡。这伴随着：（1）细胞内前列腺素（PG）E含量降低；（2）BAX和BID在细胞器膜上和细胞核内的亚细胞重新分布和聚集；（3）BAX与BID以及BAX与VDAC-1在细胞器上的共定位；（4）BCL-2聚集程度最高的细胞存活。BAX与BID以及BAX与VDAC-1的亚细胞重新分布和共定位的相似模式表明，BAX（与BID结合）控制VDAC-1的功能及其对用NIM刺激诱导凋亡的COLO 205细胞线粒体释放的凋亡因子的通透性。
