MassIVE MSV000091283

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GNPS - A systematic evaluation of quenching and extraction procedures for quantitative metabolome profiling of Hela carcinoma cell under 2D and 3D cell culture conditions

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Description

Metabolic reprogramming has been coined as a hallmark of cancer, accompanied by which the alterations in metabolite levels have profound effects on gene expression, cellular differentiation and the tumor environment. Yet a systematic evaluation of quenching and extraction procedures for quantitative metabolome profiling of tumor cells is currently lacking. To achieve this, this study is aimed at establishing an unbiased and leakage-free metabolome preparation protocol for Hela carcinoma cell. We evaluated 12 combinations of quenching and extraction methods from three quenchers (liquid nitrogen, 50% methanol, normal saline) and four extractants (80% methanol, methanol: chloroform: water (1:1:1, v/v/v), 50% acetonitrile, 70% ethanol) for global metabolite profiling of adherent Hela carcinoma cells. Based on the isotope dilution mass spectrometry (IDMS) method, gas/liquid chromatography in tandem with mass spectrometry was used to quantitatively determine 43 metabolites including sugar phosphates, organic acids, amino acids, adenosine nucleotides and coenzymes involved in central carbon metabolism. Among 12 combinations, cells that washed twice with phosphate buffered saline (PBS), quenched with liquid nitrogen, and then extracted with 50% acetonitrile was found to be the most optimal method to acquire intracellular metabolites with minimal loss during sample preparation. Furthermore, a case study was carried out to evaluate the effect of doxorubicin (DOX) on both adherent cells and 3D tumor spheroids using quantitative metabolite profiling. Based on this, quantitative time-resolved metabolite data can serve to the generation of hypotheses on metabolic reprogramming to reveal its important role in tumor development and treatment. [doi:10.25345/C5125QK54] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Hela ; metabolomics ; sample preparation ; 3D tumor spheroids ; quenching ; extraction ; isotope dilution mass spectrometry

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Principal Investigators:
(in alphabetical order) 		Wang Tong, East China University of Science and Technology, China
Submitting User: Wangtong
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GNPS content goes here (MSV000091283 [task=4a1563afd0f64d75b2992ea39c6ac493])
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Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

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