Weilbacher A, Gutekunst M, Oren M, Aulitzky W E, van der Kuip H
Dr Margarete Fischer-Bosch Institute of Clinical Pharmacology and University of Tuebingen, Auerbachstrasse 112, Stuttgart, Germany.
Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
Cell Death Dis. 2014 Jul 10;5(7):e1318. doi: 10.1038/cddis.2014.284.
Significant advances have been made in the development of small molecules blocking the p53/MDM2 interaction. The Mdm2 inhibitor Nutlin-3 is restricted to tumors carrying wtp53. In contrast, RITA, a compound that binds p53, has recently been shown also to restore transcriptional functions of mtp53. As more than 50% of solid tumors carry p53 mutations, RITA promises to be a more effective therapeutic strategy than Nutlin-3. We investigated effects of RITA on apoptosis, cell cycle and induction of 45 p53 target genes in a panel of 14 cell lines from different tumor entities with different p53 status as well as primary lymphocytes and fibroblasts. Nine cell strains expressed wtp53, four harbored mtp53, and three were characterized by the loss of p53 protein. A significant induction of cell death upon RITA was observed in 7 of 16 cell lines. The nonmalignant cells in our panel were substantially less sensitive. We found that in contrast to Nultin-3, RITA is capable to induce cell death not only in tumor cells harboring wtp53 and mtp53 but also in p53-null cells. Importantly, whereas p53 has a central role for RITA-mediated effects in wtp53 cells, neither p53 nor p63 or p73 were essential for the RITA response in mtp53 or p53-null cells in our panel demonstrating that besides the known p53-dependent action of RITA in wtp53 cells, RITA can induce cell death also independently of p53 in cells harboring defective p53. We identified an important role of both p38 and JNK/SAPK for sensitivity to RITA in these cells leading to a typical caspase- and BAX/BAK-dependent mitochondrial apoptosis. In conclusion, our data demonstrate that RITA can induce apoptosis through p38 and JNK/SAPK not only in tumor cells harboring wtp53 and mtp53 but also in p53-null cells, making RITA an interesting tumor-selective drug.
在开发阻断p53/MDM2相互作用的小分子方面已经取得了重大进展。Mdm2抑制剂Nutlin-3仅限于携带野生型p53的肿瘤。相比之下,一种与p53结合的化合物RITA最近也被证明能恢复突变型p53的转录功能。由于超过50%的实体瘤携带p53突变,RITA有望成为比Nutlin-3更有效的治疗策略。我们研究了RITA对14种来自不同肿瘤实体、具有不同p53状态的细胞系以及原代淋巴细胞和成纤维细胞中细胞凋亡、细胞周期和45个p53靶基因诱导的影响。9个细胞株表达野生型p53,4个含有突变型p53,3个以p53蛋白缺失为特征。在16个细胞系中的7个中观察到RITA诱导细胞死亡显著增加。我们实验中的非恶性细胞对RITA的敏感性明显较低。我们发现,与Nutlin-3不同,RITA不仅能够在携带野生型p53和突变型p53的肿瘤细胞中诱导细胞死亡,还能在p53缺失的细胞中诱导细胞死亡。重要的是,虽然p53在野生型p53细胞中对RITA介导的效应起核心作用,但在我们实验的突变型p53或p53缺失的细胞中,p53、p63或p73对RITA反应都不是必需的,这表明除了RITA在野生型p53细胞中已知的p53依赖性作用外,RITA在携带缺陷型p53的细胞中也能独立于p53诱导细胞死亡。我们确定了p38和JNK/SAPK在这些细胞对RITA的敏感性中都起重要作用,导致典型的半胱天冬酶和BAX/BAK依赖性线粒体凋亡。总之,我们的数据表明,RITA不仅能通过p38和JNK/SAPK在携带野生型p53和突变型p53的肿瘤细胞中诱导凋亡,还能在p53缺失的细胞中诱导凋亡,使RITA成为一种有吸引力的肿瘤选择性药物。