Synergistic induction of apoptosis by the combination of TRAIL and chemotherapy in chemoresistant ovarian cancer cells
Synergistic induction of apoptosis by the combination of TRAIL and chemotherapy in chemoresistant ovarian cancer cells
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DOI：
10.1006/gyno.2001.6194
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发表时间：
2001-06-01
影响因子：
4.7
通讯作者：
Lipkowitz, S
中科院分区：
文献类型：
--
作者：
Cuello, M;Ettenberg, SA;Lipkowitz, S
目标。本研究旨在探讨tnf相关凋亡诱导配体（TRAIL）单独或联合化疗是否能诱导化疗耐药卵巢癌细胞凋亡。12个化疗耐药上皮癌细胞系分别用每种化疗药物（顺铂、阿霉素或紫杉醇）、TRAIL单独或联合治疗。采用MTS法评估毒性。为了评估生长抑制是否由细胞凋亡引起，通过TUNEL实验、caspase激活（通过caspase-3和PARP切割测量）和亚G0/G1部分的细胞进行了测量。通过分级抑制和剂量效应分析证实了协同作用。通过免疫印迹和RNase保护实验研究死亡受体和诱饵受体的表达。均数的统计比较采用Student’st检验。大多数化疗耐药细胞也对TRAIL单独耐药。相比之下，TRAIL和化疗的结合在很大的浓度范围内导致了显著的生长抑制。通过剂量效应分析，这种相互作用是协同的。流式细胞术显示，与单独使用每种试剂相比，联合使用可显著增加凋亡细胞的比例。在联合处理后，观察到caspase和PARR切割的显著增强。最后，凋亡诱导与死亡受体水平无相关性。数据表明，几乎所有对化疗耐药的卵巢癌细胞也对TRAIL耐药，TRAIL和化疗联合通过触发caspase介导的细胞凋亡以协同方式克服这种耐药。TRAIL和化疗的结合可能是一种治疗化疗耐药卵巢癌的有效方法。(C) 2001学术出版社。
Objectives. The aim of this study was to investigate whether TNF-related apoptosis-inducing ligand (TRAIL) alone or in combination with chemotherapy could induce apoptosis in ovarian cancer cells resistant to chemotherapy,Methods. Twelve chemoresistant epithelial cancer cell lines were treated with each chemotherapeutic drug alone (cisplatin, doxorubicin, or paclitaxel), TRAIL alone, or the combination. Toxicity was assessed using the MTS assay. To assess whether growth inhibition was due to apoptosis, TUNEL assay, caspase activation (measured by caspase-3 and PARP cleavage), and the sub G0/G1 fraction of cells were measured. Synergism was confirmed by fractional inhibition and dose- effect analysis. Expression of death and decoy receptors was studied by immunoblotting and an RNase protection assay. Statistical comparison of means was performed using Student's t test.Results. The majority of the chemoresistant cells were also resistant to TRAIL alone. In contrast, the combination of TRAIL and chemotherapy resulted in a significant growth inhibition over a wide range of concentrations. This interaction was synergistic by dose-effect analysis. Flow cytometry demonstrated a significant increase in the fraction of apoptotic cells by the combination compared to each reagent alone. A significant enhancement in caspase and PARR cleavage was observed upon treatment with the combination. Finally, no correlation between induction of apoptosis and level of death receptors was found.Conclusions. The data suggest that almost all the ovarian cancer cells, which are resistant to chemotherapy, are also resistant to TRAIL, The combination of TRAIL and chemotherapy overcomes this resistance in a synergistic fashion by triggering caspase-mediated apoptosis. The combination of TRAIL and chemotherapy could be useful as a therapy for chemoresistant ovarian cancers. (C) 2001 Academic Press.