4. Conclusion
In this study, we systematically evaluated 12 combinations of quenching
and extraction methods for acquiring quantitative intracellular
metabolome with both 2D and 3D Hela carcinoma models based on the
isotope dilution mass spectrometry method. Our results showed that
liquid nitrogen as quenching agent and 50% acetonitrile as extracting
agent was proven to be the most optimal method to acquire intracellular
metabolome for both 2D Hela and 3D MTSs. Based on this, a quantitative
metabolomics study showed that DOX exposure gave rise to pronounced
metabolite changes between 2D and 3D Hela carcinoma models. Strikingly,
our data suggested that compared to 2D cells the increased intracellular
glutamine level in 3D cells can benefit replenishing the TCA cycle when
the glycolysis was limited after dosing with DOX.
In summary, this study provides a well-established quenching and
extraction protocol for quantitative metabolome profiling of Hela
carcinoma cell under 2D and 3D cell culture conditions. 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.