Jungermannenone A and B induce ROS- and cell cycle-dependent apoptosis in prostate cancer cells in vitro
Jungermannenone A and B induce ROS- and cell cycle-dependent apoptosis in prostate cancer cells in vitro
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Jungermannenone A 和 B 在体外诱导前列腺癌细胞中 ROS 和细胞周期依赖性细胞凋亡。
DOI：
10.1038/aps.2016.26
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发表时间：
2016-06-01
影响因子：
8.2
通讯作者：
Yuan, Hui-qing
中科院分区：
文献类型：
--
作者：
Guo, Yan-xia;Lin, Zhao-min;Yuan, Hui-qing
目的：Jungermannenone A和B（JA、JB）是从中国地钱Jungermannia fauriana中分离得到的新对映贝壳杉烷型二萜类化合物，具有抗癌细胞增殖活性。在这项研究中，我们研究了JA和JB的抗癌作用在PC 3人前列腺癌cells in vitro.Methods的机制：一组9人癌细胞系进行了测试。用实时细胞分析仪和MTT法评估细胞增殖。采用流式细胞仪检测细胞凋亡、细胞周期分布和活性氧水平。透射电镜观察线粒体损伤情况。彗星试验检测DNA损伤。凋亡，DNA损伤和细胞周期相关的蛋白质进行了分析，使用蛋白质印迹。结果：JA和JB对9种肿瘤细胞均有较强的抗增殖作用，其中以PC 3细胞的作用最为敏感，IC 50值分别为1.34±0.09和4.93±0.20 μmol/L。JA（1.5 μmol/L）和JB（5 μmol/L）可诱导PC 3细胞凋亡，caspase抑制剂Z-VAD可抑制JA和JB诱导的PC 3细胞凋亡。此外，JA和JB均引起PC 3细胞线粒体损伤和ROS积累，而维生素C则阻断了ROS积累并减弱了JA和JB的细胞毒性。此外，JA和JB均诱导DNA损伤，并伴有DNA修复蛋白Ku 70/Ku 80和RDA 51的下调。JA可诱导前列腺癌细胞周期阻滞于G 0/G1期，这与抑制c-Myc有关，而JB可使细胞周期阻滞于G2/M期，这与激活JNK信号通路有关。
Aim:Jungermannenone A and B (JA, JB) are new ent-kaurane diterpenoids isolated from Chinese liverwort Jungermannia fauriana, which show anti-proliferation activities in cancer cells. In this study we investigated the mechanisms underlying the anticancer action of JA and JB in PC3 human prostate cancer cells in vitro.Methods:A panel of 9 human cancer cell lines was tested. Cell proliferation was assessed with a real-time cell analyzer and MTT assay. Cell apoptosis, cell cycle distribution and ROS levels were measured using cytometry. Mitochondrial damage was examined by transmission electron microscopy. DNA damage was detected with comet assay. Apoptotic, DNA damage- and cell cycle-related proteins were analyzed using Western blotting. The expression of DNA repair genes was measured with qRT-PCR.Results:Both JA and JB exerted potent anti-proliferative action against the 9 cancer cell lines, and PC3 cells were more sensitive with IC50 values of 1.34±0.09 and 4.93±0.20 μmol/L, respectively. JA (1.5 μmol/L) and JB (5 μmol/L) induced PC3 cell apoptosis, which was attenuated by the caspase inhibitor Z-VAD. Furthermore, both JA and JB caused mitochondrial damage and ROS accumulation in PC3 cells, whereas vitamin C blocked the ROS accumulation and attenuated the cytotoxicity of JA and JB. Moreover, both JA and JB induced DNA damage, accompanied by downregulated DNA repair proteins Ku70/Ku80 and RDA51. JA induced marked cell cycle arrest at the G0/G1 phase, which was related to c-Myc suppression, whereas JB enforced the cell cycle blockade in the G2/M phase, which associated with activation of the JNK signaling.Conclusion:Both JA and JB induce prostate cancer apoptosis via ROS accumulation and induction of cell cycle arrest.