Cryptotanshinone Induces Pro-death Autophagy through JNK Signaling Mediated by Reactive Oxygen Species Generation in Lung Cancer Cells
Cryptotanshinone Induces Pro-death Autophagy through JNK Signaling Mediated by Reactive Oxygen Species Generation in Lung Cancer Cells
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DOI：
10.2174/1871520615666150907093036
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发表时间：
2016-01-01
影响因子：
2.8
通讯作者：
Chen, Xiuping
中科院分区：
文献类型：
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作者：
Hao, Wenhui;Zhang, Xuenong;Chen, Xiuping
隐丹参酮（Cryptotanshinone， CTS）是一种从丹参中分离得到的具有抗癌作用的天然物质。先前的报道显示CTS诱导caspase非依赖性细胞死亡。在这里，我们报道了CTS诱导人肺癌细胞的死亡前自噬。CTS对A549细胞的增殖具有时间和浓度依赖性。CTS触发了自噬，通过单氨尸体碱染色、透射电镜分析以及微管相关蛋白轻链3 （LC3）的western blot检测证实了这一点。CTS以浓度和时间依赖的方式诱导细胞内活性氧（ROS）的形成，这一过程被n-乙酰基- l-半胱氨酸（NAC）、过氧化氢酶、二苯乙烯（DPI）、吡罗胺二甲基硫代氨基甲酸酯（PDTC）和双豆醇逆转。NAC、JNK siRNA和SP600125抑制cts诱导的自噬。NAC逆转cts诱导的JNK磷酸化。NAC、3-甲基腺嘌呤（3-MA）和SP600125部分逆转cts诱导的细胞死亡。此外，CTS （10 mg/kg）可显著抑制A549异种移植裸鼠肿瘤生长48.3%,NAC （50 mg/kg）可完全逆转这一作用。我们的研究结果表明，CTS通过激活JNK信号，通过增加细胞内ROS的产生来诱导促死亡自噬。
Cryptotanshinone (CTS), a natural product isolated from Salvia miltiorrhiza Bunge, demonstrates anticancer effect. Previous reports showed that CTS induced caspase-independent cell death. Here, we reported that CTS induced pro-death autophagy in human lung cancer cells. CTS inhibited the proliferation of A549 cells in a time-and concentration-dependent manner. CTS triggered autophagy as confirmed by monodansylcadaverine staining, transmission electron microscopy analysis, as well as western blot detection of microtubule-associated protein light-chain 3 (LC3). CTS induced intracellular reactive oxygen species (ROS) formation in a concentration-and time-dependent manner, which was reversed by N-acetyl-L-cysteine (NAC), catalase, diphenyleneiodonium (DPI), pyrrolinodimethylthiocarbamate (PDTC), and dicumarol. Furthermore, CTS-induced autophagy was inhibited by NAC, JNK siRNA and SP600125. NAC reversed CTS-induced JNK phosphorylation. NAC, 3-methyladenine (3-MA), and SP600125 partly reversed CTS-induced cell death. In addition, CTS (10 mg/kg) dramatically inhibited tumor growth by 48.3% in A549 xenograft nude mice, which was completely reversed by NAC (50 mg/kg) co-treatment. Our findings showed that CTS induced pro-death autophagy through activating JNK signaling mediated by increasing intracellular ROS production.