Naringenin up-regulates the expression of death receptor 5 and enhances TRAIL-induced apoptosis in human lung cancer A549 cells
Naringenin up-regulates the expression of death receptor 5 and enhances TRAIL-induced apoptosis in human lung cancer A549 cells
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DOI：
10.1002/mnfr.201000024
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发表时间：
2011-02-01
影响因子：
5.2
通讯作者：
Choi, Yung Hyun
中科院分区：
文献类型：
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作者：
Jin, Cheng-Yun;Park, Cheol;Choi, Yung Hyun
范围：虽然TRAIL对正常细胞相对无毒，但它可以选择性地诱导许多类型的转化细胞凋亡。然而，一些非小细胞肺癌（NSCLC）细胞对TRAIL的作用特别耐药。在这里，我们报告说，在结合柚皮素暴露于TRAIL诱导凋亡的TRAIL耐药的NSCLC A549细胞与正常肺成纤维细胞的细胞增殖没有检测到抑制作用。采用DAPI染色和流式细胞术检测细胞凋亡。蛋白质水平通过Western印迹分析测定。使用比色测定法测量胱天蛋白酶活性。为了敲低Bid和DR5表达，通过脂质转染将Bid和DR5siRNA转染到细胞中。我们可以表明，暴露于柚皮素后，DR5蛋白被上调，并通过siRNA减弱柚皮素加TRAIL诱导的细胞凋亡来敲低DR5表达。柚皮素和TRAIL有效地诱导Bid切割，并且siRNA介导的Bid沉默降低了柚皮素的致敏作用。此外，共同治疗与柚皮素和TRAIL导致减少的克隆形成能力的A549细胞，和存活的克隆可以重新敏化重复的TRAIL treatment.Conclusion：我们的研究结果表明，治疗与TRAIL和柚皮素的组合可能是一个安全的策略，用于治疗耐药NSCLC。
Scope: While TRAIL is relatively non-toxic to normal cells, it can selectively induce apoptosis in many types of transformed cells. Nevertheless, some non-small cell lung cancer (NSCLC) cells are particularly resistant to the effects of TRAIL. Here, we report that in combination with naringenin exposure to TRAIL induced apoptosis in TRAIL-resistant NSCLC A549 cells with no detectable inhibitory effects on cell proliferation of normal lung fibroblast cells.Methods and results: Cytotoxicity was evaluated by MTT assay. Apoptosis was detected using DAPI staining, and flow cytometry. The protein levels were determined by Western blot analysis. Caspase activity was measured using a colorimetric assay. For knockdown of Bid and DR5 expression, Bid and DR5 siRNAs were transfected into cells via lipofection. We could show that following exposure to naringenin, DR5 proteins were up-regulated and knockdown of DR5 expression by siRNA attenuated naringenin plus TRAIL-induced apoptosis. Naringenin and TRAIL effectively induced Bid cleavage and siRNA-mediated silencing of Bid reduced the sensitizing effect of naringenin. Furthermore, co-treatment with naringenin and TRAIL resulted in reduction of the clonogenic capacity of A549 cells, and surviving clones could be re-sensitized for repeated TRAIL treatment.Conclusion: Our results indicate that treatment with a combination of TRAIL and naringenin may be a safe strategy for treatment of resistant NSCLC.