Green Catalysis Center, and College of Chemistry, Zhengzhou University, Zhengzhou, Henan 450001, China.
Department of Pharmacy, the First Affiliated Hospital, Zhengzhou University, Zhengzhou, Henan 450052, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Apr 15;1169:122606. doi: 10.1016/j.jchromb.2021.122606. Epub 2021 Feb 27.
Chondrosarcoma is the 2nd most frequent bone sarcoma. In this study, the metabolic profiling of human chondrosarcoma SW-1353 cell line was investigated for the first time. To obtain more precise information about the metabolites from chondrosarcoma cells, pretreatment methods including washing steps/solvents, harvesting conditions, and extraction protocols for chondrosarcoma SW-1353 cells were evaluated in the context of metabolite profiling by GC-MS technique. In addition, a total of 32 representative metabolites (related to amino acid metabolism, TCA cycle, glycolysis, and fatty acid metabolism) were quantitatively determined. We found that a fast water rinse step before metabolic quenching, may reduce the contaminants and improve sensitivity. Trypsin/ethylene diamine tetraacetic acid treatment led to a large amount of metabolite leakage, which was not suitable for metabolomics research. Methanol was selected as a more suitable extraction solvent among four extraction approaches applied to SW-1353 cells. The final protocol can provide a simple, robust, and reproducible method to obtain precise information about the metabolites from chondrosarcoma cells, which is helpful to further understand the chondrosarcoma cell physiology and the mechanism of drug resistance in this disease, from the perspective of metabolomics.
软骨肉瘤是第二常见的骨肉瘤。在这项研究中，首次对人软骨肉瘤 SW-1353 细胞系的代谢组学进行了研究。为了从软骨肉瘤细胞中获得更精确的代谢物信息，在 GC-MS 技术的代谢组学分析中，评估了包括洗涤步骤/溶剂、收获条件和软骨肉瘤 SW-1353 细胞提取方案在内的预处理方法。此外，还定量测定了 32 种具有代表性的代谢物（与氨基酸代谢、三羧酸循环、糖酵解和脂肪酸代谢有关）。我们发现，在代谢物猝灭前进行快速水冲洗步骤，可以减少污染物并提高灵敏度。胰蛋白酶/乙二胺四乙酸处理会导致大量代谢物泄漏，不适合代谢组学研究。甲醇是应用于 SW-1353 细胞的四种提取方法中更适合的提取溶剂。最终方案可以提供一种简单、稳健且可重复的方法，从代谢组学的角度获得有关软骨肉瘤细胞代谢物的精确信息，这有助于进一步了解软骨肉瘤细胞的生理学和该疾病的耐药机制。
