p53-defective tumors with a functional apoptosome-mediated pathway: A new therapeutic target
p53-defective tumors with a functional apoptosome-mediated pathway: A new therapeutic target
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DOI：
10.1093/jnci/dji133
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发表时间：
2005-05-18
影响因子：
10.3
通讯作者：
Tsuruo, T
中科院分区：
文献类型：
--
作者：
Mashima, T;Oh-Hara, T;Tsuruo, T
背景资料。尽管癌细胞似乎维持着固有的凋亡机制，但在恶性转化过程中会出现该途径的缺陷，从而阻止细胞凋亡的发生。如何特异性地诱导癌细胞的凋亡仍不清楚。方法：采用细胞色素c依赖的半胱氨酸天冬氨酸氨基转移酶(Caspase)活性检测方法和DNA测序方法，分别检测正常人细胞和肺、结肠、胃、脑、乳腺癌细胞的凋亡体活性和P53状态，并用比较分析的方法鉴定凋亡体特异性激动剂。我们比较了NCI-H23(肺癌)、HCT-15(结肠癌)和SF268(脑癌)细胞在用乙酰辅酶A合成酶(ACS)抑制剂Triacsin c处理后的细胞死亡、细胞色素c释放和caspase激活。模拟、瞬时或稳定地将抗Triacsin c的ACSL5、显性阴性的caspase-9或凋亡蛋白激活因子-1敲除的基因导入细胞。我们测量了模拟和ACSL5转导的SF268细胞的ACS活性和心磷脂(一种线粒体磷脂)的水平。携带NCI-H23移植瘤(n=10)的裸鼠接受Triacsin c或赋形剂治疗，并评估移植瘤的生长情况。组间比较采用双侧学生t检验。结果：在分析的21个p53缺陷肿瘤细胞系中，有17个细胞的凋亡体活性高于正常细胞。Triacsin c选择性诱导肿瘤细胞凋亡体介导的死亡(经Triacsin c处理的SF268细胞的caspase活性，平均值分别为1020%和100%；差值分别为920%，95%CI=900%~940%；P
Background. Although cancer cells appear to maintain the machinery for intrinsic apoptosis, defects in the pathway develop during malignant transformation, preventing apoptosis from occurring. How to specifically induce apoptosis in cancer cells remains unclear. Methods: We determined the apoptosome activity and p53 status of normal human cells and of lung, colon, stomach, brain, and breast cancer cells by measuring cytochrome c-dependent caspase activation and by DNA sequencing, respectively, and we used COMPARE analysis to identify apoptosome-specific agonists. We compared cell death, cytochrome c release, and caspase activation in NCI-H23 (lung cancer), HCT-15 (colon cancer), and SF268 (brain cancer) cells treated with Triacsin c, an inhibitor of acyl-CoA synthetase (ACS), or with vehicle. The cells were mock, transiently, or stably transfected with genes for Triacsin c-resistant ACSL5, dominant negative caspase-9, or apoptotic protease activating factor-1 knockdown. We measured ACS activity and levels of cardiolipin, a mitochondrial phospholipid, in mock and ACSL5-transduced SF268 cells. Nude mice carrying NCI-H23 xenograft tumors (n =.10) were treated with Triacsin c or vehicle, and xenograft tumor growth was assessed. Groups were compared using two-sided Student t tests. Results: Of 21 p53-defective tumor cell lines analyzed, 17 had higher apoptosome activity than did normal cells. Triacsin c selectively induced apoptosome-mediated death in tumor cells (caspase activity of Triacsin c-treated versus untreated SF268 cells; means = 1020% and 100%, respectively; difference = 920%, 95% CI = 900% to 940%; P