Metabolomics analysis reveals heavy metal copper-induced cytotoxicity in HT-29 human colon cancer cells
Metabolomics analysis reveals heavy metal copper-induced cytotoxicity in HT-29 human colon cancer cells
复制标题
代谢组学分析揭示重金属铜诱导 HT-29 人结肠癌细胞的细胞毒性
DOI：
10.1039/c6ra09320e
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发表时间：
2016
期刊：
影响因子：
3.9
通讯作者：
Chen Wei
中科院分区：
文献类型：
--
作者：
Xiao Yue;Zhai Qixiao;Wang Gang;Liu Xiaoming;Zhao Jianxin;Tian Fengwei;Zhang Hao;Chen Wei
铜 (Cu) 对人类和动物有毒，因为芬顿反应以及代谢和生物过程的异常改变会产生活性氧 (ROS)。对于普通人群来说，饮食和饮用水是铜暴露的主要来源，因此肠道是抵御铜中毒的第一道屏障。先前的研究已经证明了铜暴露的细胞毒性，重点是氧化应激和铜运输等特定方面。但有关细胞对铜暴露的整体反应的信息有限。代谢组学方法以细胞和生物体中整个代谢物的高通量检测为特色，为研究整体细胞毒性反应中的传统方法提供了有前途的替代方法。本研究选择人结肠癌细胞系HT-29，采用基于液相色谱-质谱（LC-MS）的代谢组学方法研究Cu暴露后肠细胞代谢组学的变化。结果表明，Cu处理后HT-29细胞的代谢物库发生了显着变化。总共有 77 种代谢物被鉴定为发生显着变化。通过 RT-qPCR 进行转录验证以确认代谢组学结果。通过结合代谢组学和 RT-qPCR 的数据，我们得出结论，Cu 诱导 HT-29 细胞细胞毒性的机制与诱导细胞凋亡、氧化应激增加、线粒体 β 氧化的改变以及脂质代谢和能量代谢的配置有关。我们期望这项研究为阐明铜暴露对人体肠道的毒性作用奠定基础。
Copper (Cu) is toxic to humans and animals as a result of the generation of reactive oxygen species (ROS) through Fenton reactions and abnormal alterations in metabolism and biological processes. For the general population, diet and drinking water are the main sources of Cu exposure, and the intestinal tract is therefore the first barrier against Cu-induced toxicity. Previous studies have demonstrated the cytotoxicity of Cu exposure with a focus on specific aspects such as oxidative stress and Cu transportation. But the information on the global response of cells to Cu exposure is limited. Metabolomic methods, featuring the high-flux detection of whole metabolites in cells and organisms, provide promising alternatives to conventional methods in the study of global cytotoxicity response. In this study, the human colon carcinoma cell line HT-29 was selected and a metabolomic method based on liquid chromatography-mass spectrometry (LC-MS) was adopted to investigate the alteration of the metabolomic profiles of intestinal cells after Cu exposure. The results indicated that the metabolite pools of HT-29 cells were significantly changed after Cu treatment. In total, 77 metabolites were identified as being significantly changed. Transcriptional validation by RT-qPCR was conducted to confirm metabolomic results. By combining the data from metabolomics and RT-qPCR, we concluded that the mechanisms of Cu-induced cytotoxicity in HT-29 cells were correlated with the induction of cell apoptosis, increased oxidative stress, alteration of mitochondrial β oxidation, and the configuration of lipid metabolism and energy metabolism. We expect this study to lay the foundation for the elucidation of the toxic effects of Cu exposure in the human intestinal tract.