Functional proteomics of resveratrol-induced colon cancer cell apoptosis: Caspase-6-mediated cleavage of lamin A is a major signaling loop
Functional proteomics of resveratrol-induced colon cancer cell apoptosis: Caspase-6-mediated cleavage of lamin A is a major signaling loop
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DOI：
10.1002/pmic.200500366
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发表时间：
2006-04-01
期刊：
影响因子：
3.4
通讯作者：
Pervaiz, S
中科院分区：
文献类型：
--
作者：
Lee, SC;Chan, J;Pervaiz, S
本研究探讨了白藜芦醇(RSV)诱导四种结肠癌细胞系(Bax+/-、Bax-/-、P53+/+和P53-/-)凋亡的分子基础。RSV以剂量依赖的方式诱导所有细胞的凋亡，但Bax+/-和P53+/+细胞比其基因敲除的细胞(分别为Bax-/-和P53-/-)更敏感。以Bax+/-细胞为模型，蛋白质组学分析揭示了四个RSV应答事件：Lamin A/C蛋白断裂；核糖体蛋白PO的一个更基本的等电变异体浓度增加；dUTPase和stathmin 1浓度降低。Western印迹分析证实RSV处理后Lamin A发生裂解。Caspase-6被激活，表现为caspase-6以活性形式的裂解和积累，以及蛋白酶活性的上调。RSV诱导的层蛋白A裂解和细胞凋亡可被caspase-6的多肽抑制剂完全阻断。同样，使用小干扰RNA部分抑制caspase-6的表达，可显著抑制RSV诱导的层蛋白A的切割和细胞凋亡。此外，基因敲除细胞(Bax-/-和P53-/-)的较低的凋亡敏感性与caspase-6和lamin A裂解的相对减少有关。综上所述，这些数据强调了caspase-6及其对lamin A的裂解在RSV在结肠癌细胞中触发的凋亡信号中的关键作用，在没有Bax或P53的情况下可以激活该信号。
The study investigated the molecular basis of resveratrol (RSV)-evoked apoptosis in four (Bax+/-, Bax-/-, p53+/+, and p53-/-) HCT116 colon cancer cell lines. RSV induced apoptosis in all the cells in a dose-dependent manner; however, Bax+/- and p53+/+ cells were more susceptible than their knockout counterparts (Bax-/- and p53-/-, respectively). Using Bax+/- cells as a model, proteomic analysis revealed four RSV-responsive events: fragmentation of lamin A/C protein; increase in concentration of a more basic isoelectric variant of the ribosomal protein PO; and decrease in concentration of dUTPase as well as stathmin 1. Lamin A cleavage in response to RSV treatment was confirmed using Western blot analysis. Caspase-6 was activated, which was evidenced by cleavage and accumulation in active form of caspase-6 as well as upregulation of the protease activity. RSV-elicited lamin A cleavage and apoptosis were entirely abrogated by the peptide inhibitors of caspase-6. Likewise, partial knockdown of caspase-6 expression using small interfering RNA resulted in significant inhibition of RSV-elicited lamin A cleavage and apoptosis. Furthermore, the lower apoptosis sensitivity of the knockout cells (Bax-/- and p53-/-) correlated with the relatively reduced processing of caspase-6 and lamin A cleavage. Taken together, these data highlight the critical role of caspase-6 and its cleavage of lamin A in apoptotic signaling triggered by RSV in the colon carcinoma cells, which can be activated in the absence of Bax or p53.