(E)-2,4-bis(p-hydroxyphenyl)-2-butenal has an antiproliferative effect on NSCLC cells induced by p38 MAPK-mediated suppression of NF-κB and up-regulation of TNFRSF10B (DR5)
(E)-2,4-bis(p-hydroxyphenyl)-2-butenal has an antiproliferative effect on NSCLC cells induced by p38 MAPK-mediated suppression of NF-κB and up-regulation of TNFRSF10B (DR5)
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DOI：
10.1111/bph.12024
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发表时间：
2013-03-01
影响因子：
7.3
通讯作者：
Hong, Jin Tae
中科院分区：
文献类型：
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作者：
Kollipara, Pushpa Saranya;Jeong, Heon Sang;Hong, Jin Tae
背景和目的美拉德反应产物（Maillard Reaction Products，MRPs）是一种有效的化学预防剂。本文主要研究了（E）-2，4-双（对羟基苯基）-2-丁烯醛（MRP）对人非小细胞肺癌（NSCLC）细胞的抗肿瘤作用及其作用机制。实验方法我们通过使用蛋白质印迹分析主要凋亡蛋白、MAPK、NF-B和死亡受体表达来分析（E）-2，4-双（对羟基苯基）-2-丁烯醛对NSCLC细胞（NCI-H460和A549）的活性。我们还用RT-PCR测定了它对死亡受体mRNA表达的影响，EMSA测定了它对NF-B DNA结合活性的影响，并用集落形成试验测定了抑制剂对（E）-2，4-双（对羟基苯基）-2-丁烯醛作用的影响。关键结果（E）-2，4-双（对羟基苯基）-2-丁烯醛因诱导细胞凋亡而对非小细胞肺癌细胞的生长产生浓度（1040 g中心点mL 1）和时间（30分钟72小时）依赖性抑制作用。同时，它显着增加凋亡蛋白的表达，如裂解的caspase-3，裂解的caspase-9，Bax和p53，但下调抗凋亡蛋白Bcl-2，cIAP 1和cIAP 2的表达。这种作用是通过MAPK和死亡受体蛋白TNFRSF 12、TNFRSF 10 B和TNFRSF 21的上调而NF-B的抑制诱导的。在激活的死亡受体中，只有用siRNA敲低TNFRSF 10 B逆转了（E）-2，4-双（对羟基苯基）-2-丁烯醛的作用。尽管所有的MAPK都被激活，但只有用p38 MAPK抑制剂预处理才能逆转（E）-2，4-双（对羟基苯基）-2-丁烯醛诱导的细胞生长抑制、裂解的半胱天冬酶-3，9和TNFRSF 10 B表达的增加以及NF-B失活。结论与意义（E）-2，4-bis（p-hydroxyphenyl）-2-butenal通过p38 MAPK介导的NF-B抑制和TNFRSF 10 B激活，进而激活caspase-3和caspase-9通路，诱导NSCLC细胞凋亡。
Background And Purpose The Maillard Reaction Products (MRPs) are known to be effective in chemoprevention. Here we focused on the anticancer effects of (E)-2,4-bis(p-hydroxyphenyl)-2-butenal (a MRP) on human non-small-cell lung cancer (NSCLC) cells and its mechanism of action. Experimental Approach We analysed the activity of (E)-2,4-bis(p-hydroxyphenyl)-2-butenal on NSCLC cells (NCI-H460 and A549) by use of Western blot analysis for major apoptotic proteins, MAPK, NF-B and death receptor expression. We also used RT-PCR to determine its effects on death receptor mRNA expression, EMSA for effects on NF-B DNA binding activity and colony formation assay for effects of inhibitors on (E)-2,4-bis(p-hydroxyphenyl)-2-butenal's actions. Key Results (E)-2,4-bis(p-hydroxyphenyl)-2-butenal induced a concentration (1040g center dot mL1)- and time (30min72h)-dependent inhibitory effect on the growth of NSCLC cells due to induction of apoptosis. Concomitantly, it significantly increased the expression of apoptotic proteins such as cleaved caspase-3, cleaved caspase-9, Bax and p53, but down-regulated the expression of anti-apoptotic proteins Bcl-2, cIAP1 and cIAP2. This effect was induced by up-regulation of MAPK and death receptor proteins TNFRSF12, TNFRSF10B and TNFRSF21, but suppression of NF-B. Of the death receptors activated, only TNFRSF10B knock down with siRNA reversed the effect of (E)-2,4-bis(p-hydroxyphenyl)-2-butenal. Even though all the MAPKs were activated, only pretreatment with a p38 MAPK inhibitor reversed (E)-2,4-bis(p-hydroxyphenyl)-2-butenal-induced cell growth inhibition, increase in cleaved caspase-3, -9 and TNFRSF10B expression, and NF-B inactivation. Conclusions And Implications (E)-2,4-bis(p-hydroxyphenyl)-2-butenal induces apoptosis in NSCLC cells by p38 MAPK-mediated suppression of NF-B and activation of TNFRSF10B, which then activates the caspase-3 and caspase-9 pathways.