Conclusion
In this study, based on the liquid
overlay method, we established a rapid and efficient method to produce
uniform size and high reproducibility of 3D MTSs.
Based on multi-omics data, the
phenotypic differences of HeLa cells under different culture conditions
were explored based on the established 3D MTS model to investigate the
resistance mechanism of 5-FU and discover the key regulatory genes and
related reprogrammed metabolic pathways. We found that in 3D HeLa
carcinoma cells the metabolic shift towards glycolysis, the maintenance
of ER homeostasis and the up-regulation of ECM may contribute to the
formation of increased resistance to 5-FU. In addition, the changes in
ABC transporters, HIF-1 signaling pathway and
the improvement of tumor cell
stemness were also related to the increase of 5-FU resistance of 3D
MTSs. Overall, this study demonstrates that resistant 3D Hela MTS can be
a promising tool for testing drug delivery and efficacy for cervical
cancer treatment, and a multi-omics analysis can reveal metabolic
addictions which can be manipulated to restore susceptibility to
chemotherapy drugs, since this model is readily compatible with the
methodologies and techniques that are already used for the analysis of
conventional 2D monolayer cultures.