Matrine induces RIP3-dependent necroptosis in cholangiocarcinoma cells.
Matrine induces RIP3-dependent necroptosis in cholangiocarcinoma cells.
复制标题
苦参碱诱导胆管癌细胞 RIP3 依赖性坏死性凋亡
DOI：
10.1038/cddiscovery.2016.96
复制
发表时间：
2017
影响因子：
7
通讯作者：
Hu T
中科院分区：
文献类型：
--
作者：
Xu B;Xu M;Tian Y;Yu Q;Zhao Y;Chen X;Mi P;Cao H;Zhang B;Song G;Zhan YY;Hu T
对促凋亡抗肿瘤药物产生获得性耐药性是胆管癌（CCA）治愈的主要障碍。诱导非凋亡细胞死亡的抗肿瘤药物被认为是克服这种耐药性的新方法。在这里，我们首次报道了苦参碱在 CCA 细胞系中诱导的坏死性凋亡，这与其在许多其他种类的癌细胞中诱导细胞凋亡的经典作用不同。苦参碱处理下的CCA细胞表现出典型的坏死样但非凋亡的形态学变化。这些苦参碱诱导的 CCA 细胞形态变化和细胞死亡可被坏死性凋亡抑制剂 necrostatin-1 大大减弱，但不能被凋亡抑制剂 z-VAD-fmk 减弱。与许多受体相互作用蛋白 3 (RIP3) 负表达的癌细胞不同，CCA 细胞中观察到 RIP3 中等表达，并且是苦参碱诱导坏死性凋亡所必需的，在敲低内源性 RIP3 后，坏死性凋亡转变为细胞凋亡。此外，苦参碱可以增加RIP3的表达水平，这可能促进坏死性凋亡过程。作为 RIP3 的下游事件，混合谱系激酶结构域 (MLKL) 从细胞质易位到质膜，以及 RIP3/MLKL 增加活性氧 (ROS) 的产生，对于苦参碱诱导坏死性凋亡至关重要。在临床研究中，我们发现与邻近正常组织相比，RIP3在大多数CCA组织样本中表达较低，但仍中等表达。综上所述，我们通过增强 RIP3 表达和随后的 RIP3/MLKL/ROS 信号通路，确定苦参碱是 CCA 中的坏死性凋亡诱导剂，这为克服 CCA 治疗中的化疗耐药性提供了基于 RIP3 表达的新的个体化策略。
The development of acquired resistance to pro-apoptotic antitumor agents is a major impediment to the cure of cholangiocarcinoma (CCA). Antitumor drugs inducing non-apoptotic cell death are considered as a new approach to overcome such drug resistance. Here, we reported for the first time that matrine-induced necroptosis in CCA cell lines, differing from its classical role to induce apoptosis in many other kinds of cancer cells. CCA cells under matrine treatment exhibited typical necrosis-like but not apoptotic morphologic change. These matrine-induced morphologic change and cell death in CCA cells were greatly attenuated by necroptosis inhibitor necrostatin-1, but not apoptosis inhibitor z-VAD-fmk. Unlike many cancer cells with negative receptor-interacting protein 3 (RIP3) expression, moderate expression of RIP3 in CCA cells was observed and was required for matrine to induce necroptosis, which was switched to apoptosis after knocking down endogenous RIP3. Moreover, matrine could increase RIP3 expression level, which may facilitate the necroptosis process. Translocation of mixed lineage kinase-domain like (MLKL) from cytoplasm to plasma membrane as a downstream event of RIP3, as well as the increased production of reactive oxygen species (ROS) by RIP3/MLKL, was critical for matrine to induce necroptosis. In clinical study, we found RIP3 was lower but still moderately expressed in most CCA tissue samples compared with adjacent normal tissues. Taken together, we identified matrine as a necroptosis inducer in CCA by enhancing RIP3 expression and the following RIP3/MLKL/ROS signaling pathway, which provided new individualized strategies based on RIP3 expression to overcome chemoresistance in CCA therapy.