Mitochondria as the primary target of resveratrol-induced apoptosis in human retinoblastoma cells
Mitochondria as the primary target of resveratrol-induced apoptosis in human retinoblastoma cells
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DOI：
10.1167/iovs.06-0119
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发表时间：
2006-09-01
影响因子：
4.4
通讯作者：
Polans, Arthur S.
中科院分区：
文献类型：
--
作者：
Sareen, Dhruv;van Ginkel, Paul R.;Polans, Arthur S.
目的.研究白藜芦醇诱导视网膜母细胞瘤细胞死亡的分子机制。白藜芦醇处理后，Y 79肿瘤细胞的生存能力进行了测量，使用基于荧光的测定，并通过Hoechst染色和流式细胞术，分别表征了促凋亡和抗增殖作用。使用5，5 '，6，6'-四氯-1，1 '，3，3'-四乙基-苯并咪唑羰花青碘化物（JC-1）测量作为药物处理的函数的线粒体跨膜电位（Delta Psi（m）），而通过免疫印迹法测定细胞色素c从线粒体的释放，并通过监测荧光肽底物的切割来测定半胱天冬酶活性。白藜芦醇诱导Y 79肿瘤细胞活力的剂量和时间依赖性降低，并通过诱导S期生长停滞和凋亡细胞死亡来抑制增殖。在细胞死亡之前，白藜芦醇引起Delta Psi（m）的快速消散。随后细胞色素c释放到细胞质中，caspase-9和caspase-3的活性大幅增加。此外，在无细胞体系中，白藜芦醇直接诱导离体线粒体去极化。结论。这些结果表明，白藜芦醇，一种无毒的天然植物化合物，抑制Y 79细胞增殖，并通过激活线粒体（内在）凋亡途径刺激凋亡，并可能需要进一步探索作为常规抗癌疗法的辅助视网膜母细胞瘤。
PURPOSE. To determine the molecular mechanisms by which resveratrol induces retinoblastoma tumor cell death.METHODS. After resveratrol treatment, Y79 tumor cell viability was measured using a fluorescence-based assay, and proapoptotic and antiproliferative effects were characterized by Hoechst stain and flow cytometry, respectively. Mitochondrial transmembrane potential ( Delta Psi(m)) was measured as a function of drug treatment using 5,5', 6,6'-tetrachloro-1,1', 3,3'-tetraethyl-benzamidazolocarbocyanin iodide ( JC-1), whereas the release of cytochrome c from mitochondria was assayed by immunoblotting and caspase activities were determined by monitoring the cleavage of fluorogenic peptide substrates.RESULTS. Resveratrol induced a dose- and time-dependent decrease in Y79 tumor cell viability and inhibited proliferation by inducing S-phase growth arrest and apoptotic cell death. Preceding cell death, resveratrol evoked a rapid dissipation of Delta Psi(m). This was followed by the release of cytochrome c into the cytoplasm and a substantial increase in the activities of caspase-9 and caspase-3. Additionally, in a cell-free system, resveratrol directly induced the depolarization of isolated mitochondria.CONCLUSIONS. These results demonstrate that resveratrol, a nontoxic natural plant compound, inhibits Y79 cell proliferation and stimulates apoptosis through activation of the mitochondrial ( intrinsic) apoptotic pathway and may warrant further exploration as an adjuvant to conventional anticancer therapies for retinoblastoma.