Emodin azide methyl anthraquinone derivative triggers mitochondrial-dependent cell apoptosis involving in caspase-8-mediated Bid cleavage
Emodin azide methyl anthraquinone derivative triggers mitochondrial-dependent cell apoptosis involving in caspase-8-mediated Bid cleavage
复制标题
大黄素叠氮甲基蒽醌衍生物触发线粒体依赖性细胞凋亡，涉及 caspase-8 介导的 Bid 裂解
DOI：
10.1158/1535-7163.mct-07-2362
复制
发表时间：
2008-06-01
影响因子：
5.7
通讯作者：
Fu, Liwu
中科院分区：
文献类型：
--
作者：
Yan, Yanyan;Su, Xiaodong;Fu, Liwu
AMAD是从天然中药虎杖中提取的大黄素叠氮甲基蒽醌衍生物。在这里，我们研究了AMAD诱导人乳腺癌细胞系MDA-MB-453和人肺腺癌Calu-3细胞凋亡的抗癌活性和信号通路。发现AMAD对两种细胞系都具有强效细胞毒性作用。Hoechst 33258染色和Annexin V/碘化丙啶双染色显示典型的凋亡细胞核特征，Annexin V阳性细胞比例呈剂量依赖性增加。此外，这种凋亡诱导与线粒体膜电位的崩溃和激活的半胱氨酸天冬氨酸蛋白酶（半胱氨酸天冬氨酸蛋白酶）级联反应，涉及半胱氨酸天冬氨酸蛋白酶-8，9，半胱氨酸天冬氨酸蛋白酶-3，和聚（ADP-核糖）聚合酶裂解在浓度依赖性的方式。值得注意的是，AMAD还有效地切割Bid，一种含有BH 3结构域的促凋亡Bcl-2家族成员，并诱导细胞色素c随后从线粒体释放到细胞质中。此外，用Z-IETD-FMK抑制caspase-8活性部分抑制了AMAD诱导的细胞色素c释放和Bid裂解，而暴露于caspase-9抑制剂Z-LETD-FMK则没有影响。此外，AMAD触发的其他线粒体膜蛋白，如Bcl-xl和Bad也发生了显著变化。有趣的是，AMAD减少了两种细胞系中活性氧的产生。DNA结合实验表明AMAD诱导的细胞凋亡不参与DNA嵌入。总之，这些数据表明，AMAD通过线粒体途径诱导细胞凋亡，涉及两种细胞系中的caspase-8/Bid活化。
AMAD, an emodin azide methyl anthraquinone derivative, was extracted from the nature giant knotweed rhizome of traditional Chinese herbs. Here, we investigated the anticancer activities and signaling pathways implicated in AMAD-induced apoptosis in human breast cancer cell lines MDA-MB-453 and human lung adenocarcinoma Calu-3 cells. AMAD was found to have a potent cytotoxic effect on both cell lines. Hoechst 33258 staining and Annexin V/propidium iodide double staining exhibited the typical nuclear features of apoptosis and increased the proportion of apoptotic Annexin V-positive cells in a dose-dependent manner, respectively. Moreover, this apoptotic induction was associated with a collapse of the mitochondrial membrane potential and activated caspases (cysteine aspartase) cascade involving in caspase-8, caspase-9, caspase-3, and poly(ADP-ribose) polymerase cleavage in a concentration-dependent manner. It was noteworthy that AMAD also effectively cleaved Bid, a BH3 domain-containing proapoptotic Bcl-2 family member, and induced the subsequent release of cytochrome c from mitochondria into the cytosol. Furthermore, suppression of caspase-8 activity with Z-IETD-FMK partially inhibited release of cytochrome c and Bid cleavage induced by AMAD, whereas exposure to Z-LETD-FMK, a caspase-9 inhibitor, had no effect. Additionally, there was significant change in other mitochondrial membrane proteins triggered by AMAD, such as Bcl-xl and Bad. It was intriguing that AMAD decreased the generation of reactive oxygen species in both cell lines. DNA-binding assay exhibited apoptosis induced by AMAD was not involved in intercalating to DNA. Taken together, these data suggested that AMAD induced apoptosis via a mitochondrial pathway involving caspase-8/Bid activation in both cell lines.