A TrxR inhibiting gold(I) NHC complex induces apoptosis through ASK1-p38-MAPK signaling in pancreatic cancer cells.
A TrxR inhibiting gold(I) NHC complex induces apoptosis through ASK1-p38-MAPK signaling in pancreatic cancer cells.
复制标题
DOI：
10.1186/1476-4598-13-221
复制
发表时间：
2014-09-25
期刊：
影响因子：
37.3
通讯作者：
Wölfl S
中科院分区：
文献类型：
--
作者：
Cheng X;Holenya P;Can S;Alborzinia H;Rubbiani R;Ott I;Wölfl S
晚期癌细胞表现出异常的抗氧化能力，可以解毒过量的ROS，从而补偿内在的氧化应激和治疗抵抗。 PDAC 是最致命的癌症之一，通常与 ROS 的大量积累有关。最近的研究发现金 (I) NHC 复合物是有效的 TrxR 抑制剂，可以在低微摩尔浓度下抑制多种人类恶性细胞系的细胞生长。然而，其作用机制尚未完全阐明。为了了解金 (I) NHC 复合物在 PDAC 中的生物学功能，我们使用了最近发表的金 (I) NHC 复合物 MC3，并通过 MTT 和 SRB 测定评估了其在四种 PDAC 细胞系中的抗增殖作用。在进一步详细分析中，我们使用 ROS 指示剂 DHE 和 Panc1 中染料 JC-1 指示的线粒体膜电位分析了细胞 ROS 水平。我们还通过 FACS 分析了细胞周期停滞和细胞凋亡。为了阐明特定细胞信号通路在 MC3 诱导的细胞死亡中的作用，与 ROS 清除剂、p38-MAPK 抑制剂和 siRNA 介导的 ASK1 耗竭进行共孵育，并通过免疫印迹、ELISA 微阵列、qRT-PCR 和免疫沉淀分析结果。我们的数据表明，MC3 有效抑制胰腺癌细胞的细胞生长，并诱导细胞周期停滞和细胞凋亡，特别是在吉西他滨耐药的癌细胞 Panc1 和 ASPC1 中。 MC3 治疗导致细胞氧化还原稳态发生显着改变，导致 ROS 水平升高和线粒体膜电位降低。 ROS 清除剂抑制 ROS 形成并挽救细胞免受损伤。在分子水平上，MC3 阻断 Trx 与 ASK1 的相互作用，随后激活 p38 相关信号传导。此外，使用 ASK1 siRNA 或 p38 抑制剂抑制该途径可明显减弱 MC3 对 Panc1 和 ASPC1 细胞增殖的影响。我们的结果证实 MC3 是一种 TrxR 抑制剂，并显示 MC3 诱导吉西他滨耐药 PDAC 细胞凋亡。使用抗氧化剂、ASK1 siRNA 或 p38 抑制剂可以阻断 MC3 介导的细胞死亡，表明 Trx-ASK1-p38 信号级联在金 (I) NHC 复合物介导的细胞损伤中发挥重要作用。本文的在线版本 (doi:10.1186/1476-4598-13-221) 包含补充材料，可供授权用户使用。
Cancer cells in the advanced stage show aberrant antioxidant capacity to detoxify excessive ROS resulting in the compensation for intrinsic oxidative stress and therapeutic resistance. PDAC is one of the most lethal cancers and often associated with a high accumulation of ROS. Recent studies identified gold(I) NHC complexes as potent TrxR inhibitors suppressing cell growth in a wide spectrum of human malignant cell lines at the low micromolar concentration. However, the mechanism of action is not completely elucidated yet. To understand the biological function of gold(I) NHC complexes in PDAC, we used a recently published gold(I) NHC complex, MC3, and evaluated its anti-proliferative effect in four PDAC cell lines, determined by MTT and SRB assays. In further detailed analysis, we analyzed cellular ROS levels using the ROS indicator DHE and mitochondrial membrane potential indicated by the dye JC-1 in Panc1. We also analyzed cell cycle arrest and apoptosis by FACS. To elucidate the role of specific cell signaling pathways in MC3-induced cell death, co-incubation with ROS scavengers, a p38-MAPK inhibitor and siRNA mediated depletion of ASK1 were performed, and results were analyzed by immunoblotting, ELISA-microarrays, qRT-PCR and immunoprecipitation. Our data demonstrate that MC3 efficiently suppressed cell growth, and induced cell cycle arrest and apoptosis in pancreatic cancer cells, in particular in the gemcitabine-resistant cancer cells Panc1 and ASPC1. Treatment with MC3 resulted in a substantial alteration of the cellular redox homeostasis leading to increased ROS levels and a decrease in the mitochondrial membrane potential. ROS scavengers suppressed ROS formation and rescued cells from damage. On the molecular level, MC3 blocked the interaction of Trx with ASK1 and subsequently activated p38-associated signaling. Furthermore, inhibition of this pathway by using ASK1 siRNA or a p38 inhibitor clearly attenuated the effect of MC3 on cell proliferation in Panc1 and ASPC1. Our results confirm that MC3 is a TrxR inhibitor and show MC3 induced apoptosis in gemcitabine-resistant PDACs. MC3 mediated cell death could be blocked by using anti-oxidants, ASK1 siRNA or p38 inhibitor suggesting that the Trx-ASK1-p38 signal cascade played an important role in gold(I) NHC complexes-mediated cellular damage. The online version of this article (doi:10.1186/1476-4598-13-221) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
3.7
作者：
Alborzinia H;Can S;Holenya P;Scholl C;Lederer E;Kitanovic I;Wölfl S
通讯作者：
Wölfl S
影响因子：
4.7
作者：
Kong, Q;Lillehei, KO
通讯作者：
Lillehei, KO
影响因子：
50.3
作者：
Dolado, Ignacio;Swat, Aneta;Nebreda, Angel R.
通讯作者：
Nebreda, Angel R.
影响因子：
50.3
作者：
Hingorani, SR;Wang, LF;Tuveson, DA
通讯作者：
Tuveson, DA
影响因子：
6.4
作者：
BERROZPE, G;SCHAEFFER, J;PERUCHO, M
通讯作者：
PERUCHO, M