Metabolite profiling of chondrosarcoma cells: A robust GC-MS method for the analysis of endogenous metabolome

Abstract

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.