Solanine-induced reactive oxygen species inhibit the growth of human hepatocellular carcinoma HepG2 cells
Solanine-induced reactive oxygen species inhibit the growth of human hepatocellular carcinoma HepG2 cells
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茄碱诱导的活性氧抑制人肝癌HepG2细胞的生长
DOI：
10.3892/ol.2016.4167
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发表时间：
2016-03-01
期刊：
影响因子：
2.9
通讯作者：
Zheng, Shu-Sen
中科院分区：
文献类型：
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作者：
Meng, Xue-Qin;Zhang, Wei;Zheng, Shu-Sen
本研究旨在探讨龙葵碱对人肝癌HepG 2细胞产生活性氧（ROS）的影响，并探讨其诱导肿瘤细胞凋亡的分子机制。在体外将龙葵碱施用至HepG 2细胞。选择靶向ROS的各种细胞定位的探针用于使用流式细胞术检测ROS表达。Western blotting检测凋亡相关蛋白（凋亡信号调节激酶1（ASK 1）、硫氧还蛋白结合蛋白2（TBP-2））和增殖相关蛋白（组蛋白去乙酰化酶1（HDAC 1））的表达水平。使用膜联蛋白V-异硫氰酸荧光素/碘化丙啶测定法测量经历凋亡的细胞的百分比，并使用瑞氏染色法检查细胞形态，然后进行倒置显微镜分析。ROS检测探针2，7-二氯荧光素二乙酸酯和二氢罗丹明123检测发现，与对照组相比，茄碱处理后的HepG 2细胞胞浆和线粒体中产生了大量的ROS，包括羟自由基（OH-）和过氧化氢（H2 O2）（P0. 05）。Western blotting结果显示龙葵碱上调ASK 1和TBP-2的表达水平，并增强其激酶活性，而龙葵碱降低增殖相关蛋白HDAC 1的表达水平。细胞凋亡率明显增加（P
The aim of the present study was to investigate the effect of solanine on promoting human hepatocellular carcinoma HepG2 cells to produce reactive oxygen species (ROS), and the molecular mechanisms leading to tumor cell apoptosis. Solanine was administered to HepG2 cells in vitro. A selection of probes targeting various cellular localizations of ROS were used to detect ROS expression using flow cytometry. The expression levels of apoptosis-associated proteins, including apoptosis signal-regulating kinase 1 (ASK1) and thioredoxin binding protein 2 (TBP-2), and proliferation-associated proteins, including histone deacetylase 1 (HDAC1), were detected using western blotting. The percentage of cells undergoing apoptosis was measured using an Annexin V-fluorescein isothiocyanate/propidium iodide assay, and cell morphology was examined using Wright's stain followed by inverted microscopy analysis. ROS detection probes 2,7-dichlorofluorescin diacetate and dihydrorhodamine 123 identified that abundant ROS, including hydroxyl radical (OH-) and hydrogen peroxide (H2O2), were produced in the cytoplasm and mitochondria of the solanine-treated HepG2 cells compared with the control cells (P0.05). Western blotting results revealed that solanine upregulated the expression levels of ASK1 and TBP-2 and enhanced their kinase activities, whereas solanine decreased the expression level of the proliferation-associated protein, HDAC1. The cell apoptotic rate was significantly increased (P