The shift in cellular metabolism, also called Warburg effect, is an emerging hallmark of cancer cells (CC) orchestrated by oncogenic proteins. MIF, a pleiotropic cytokine, is involved in CC proliferation and multiple aspects of carcinogenesis but little is known about its possible role in cellular metabolic activities. Using MIF knockdown (KD) technique and 1H-RMN spectroscopy, we evaluated the influence of MIF on MDA-MB-231 and MCF-7cells.
MIF KD cells were obtained by lentiviral transduction. MDA-MB-231 MIF sc/KD and MCF-7 MIF sc/KD cells were scrapped and quenched using methanol. The solution containing the quenched cells was pipetted for intracellular metabolites extraction (methanol, chloroform and water extraction procedure). Only the aqueous phase was used. Each sample was reconstituted in phosphate buffer mixed with TSP. After acquisition on a Bruker Avance spectrometer at 500 MHz, NMR spectra are integrated in 0.04ppm subregion (binning). Multivariate analysis (Partial Least Square Discriminate Analysis [PLS-DA]) was performed on the integrated data (SIMCA P + , MKS Data Analytics Solutions).
PLS-DA analysis of NMR data clearly separate the MDA-MB-231 from MCF-7 cells (ANOVA, p
Our data show that MDA-MB-231 and MCF-7 cells have different metabolic profiles which suggest that MDA-MB-231 cells are more dependent on glutamine but less on lactate. The decrease of glutamine levels observed in both cells lines when MIF is under-expressed suggests that MIF could be involved in the regulation of this metabolite.
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All authors have declared no conflicts of interest.