The Fuzhou Second Hospital Affiliated to Xiamen University, Fuzhou, 350007, China; Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, MOE Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing, 100084, China.
Department of Chemistry, Beijing Key Laboratory of Microanalytical Methods and Instrumentation, MOE Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing, 100084, China.
Talanta. 2019 Nov 1;204:6-12. doi: 10.1016/j.talanta.2019.05.088. Epub 2019 May 23.
Cancer stem cells (CSCs) are the origin of many malignant tumours, including osteosarcoma that mainly affects adolescents and is accompanied by a poor prognosis. However, little is known about the intrinsic biological information of osteosarcoma stem cells, particularly for the metabolomics features. Hence, an ultra-high performance liquid chromatography coupled with tandem Q-Exactive Orbitrap mass spectrometer (UHPLC-QE-MS)-based metabolomics approach was developed to investigate the metabolism changes in the human osteosarcoma (HOS) cell line in order to understand its possible mechanism. HMDB, METLIN and m/z Cloud databases were used to identify the metabolic markers. Additionally, the compounds were further identified using standards of the metabolites. Comparing HOS-CSCs with non-CSCs, 154 different metabolites were identified in both the positive and negative modes. Based on the clearly distinct metabolites, the changed metabolic pathways were analysed using MetaboAnalyst. The top five altered pathways included alanine, aspartate and glutamate metabolism; arginine and proline metabolism; glutathione metabolism; cysteine and methionine metabolism; and the citrate cycle (TCA cycle). The downregulation of the TCA cycle and elevation of oxidized glutathione levels suggested a decline of mitochondrial metabolism, while most of the amino acid metabolisms were upregulated. Further biological experiments including an analysis of mitochondrial activity confirmed the above hypotheses that were deduced from metabolomics results. These findings not only enhance our understanding of the altered metabolome in osteosarcoma stem cells but also demonstrate the great potential of such a metabolomics method based on UHPLC-QE-MS in large-scale cell studies.
癌症干细胞（CSC）是许多恶性肿瘤的起源，包括主要影响青少年且预后不良的骨肉瘤。然而，对于骨肉瘤干细胞的内在生物学信息，特别是代谢组学特征，了解甚少。因此，开发了一种基于超高效液相色谱串联 Q-Exactive Orbitrap 质谱仪（UHPLC-QE-MS）的代谢组学方法，以研究人骨肉瘤（HOS）细胞系的代谢变化，以了解其可能的机制。使用 HMDB、METLIN 和 m/z Cloud 数据库来鉴定代谢标志物。此外，使用代谢物的标准品进一步鉴定化合物。将 HOS-CSCs 与非-CSCs 进行比较，在正、负离子模式下共鉴定出 154 种不同的代谢物。基于明显不同的代谢物，使用 MetaboAnalyst 分析了改变的代谢途径。前五个改变的途径包括丙氨酸、天冬氨酸和谷氨酸代谢；精氨酸和脯氨酸代谢；谷胱甘肽代谢；半胱氨酸和蛋氨酸代谢；以及柠檬酸循环（TCA 循环）。TCA 循环的下调和氧化型谷胱甘肽水平的升高表明线粒体代谢的下降，而大多数氨基酸代谢物上调。进一步的生物学实验，包括线粒体活性分析，证实了上述从代谢组学结果推断出的假设。这些发现不仅增强了我们对骨肉瘤干细胞中改变的代谢组的理解，还展示了基于 UHPLC-QE-MS 的代谢组学方法在大规模细胞研究中的巨大潜力。
