Metabolic profiling of attached and detached metformin and 2-deoxy-D-glucose treated breast cancer cells reveals adaptive changes in metabolome of detached cells

Abstract

Anchorage-independent growth of cancer cells in vitro is correlated to metastasis formation in vivo. Metformin use is associated with decreased breast cancer incidence and currently evaluated in cancer clinical trials. The combined treatment with metformin and 2-deoxy-D-glucose (2DG) in vitro induces detachment of viable MDA-MB-231 breast cancer cells that retain their proliferation capacity. This might be important for cell detachment from primary tumors, but the metabolic changes involved are unknown. We performed LC/MS metabolic profiling on separated attached and detached MDA-MB-231 cells treated with metformin and/or 2DG. High 2DG and metformin plus 2DG altered the metabolic profile similarly to metformin, inferring that metabolic changes are necessary but not sufficient while the specific effects of 2DG are crucial for detachment. Detached cells had higher NADPH levels and lower fatty acids and glutamine levels compared to attached cells, supporting the role of AMPK activation and reductive carboxylation in supporting anchorage-independent survival. Surprisingly, the metabolic profile of detached cells was closer to untreated control cells than attached treated cells, suggesting detachment might help cells adapt to energy stress. Metformin treated cells had higher fatty and amino acid levels with lower purine nucleotide levels, which is relevant for understanding the anticancer mechanisms of metformin.

Dual Effect of Combined Metformin and 2-Deoxy-D-Glucose Treatment on Mitochondrial Biogenesis and PD-L1 Expression in Triple-Negative Breast Cancer Cells

“…Additionally, PD-L1 levels in Met + 0.6DG treated cells were normalized by mannose while the AMPK inhibitor compound C had no such effect ( Figure 7 F). We speculate that metformin treatment potentiates the effect of 2DG on protein N-glycosylation by depleting the pools of glycolysis intermediates, as observed in our previous work [ 54 ].…”

“…As the main in vitro model, we therefore used two TNBC cell lines (MDA-MB-231 and BT-549) resistant to anoikis for which we have observed the phenomenon of detachment and survival in anchorage-independent conditions after treatment with metformin + 0.6 mM 2DG (Met + 0.6DG) or 4.8 mM 2DG (4.8DG) [ 16 , 30 , 54 ]. This could be especially relevant in the nutrient-poor tumor microenvironment, so we also analyzed the effects of metformin and 2DG in the absence of glucose.…”

Regulating TKT activity inhibits proliferation of human acute lymphoblastic leukemia cells

“…Another glucose analog, 2-deoxyglucose (2 -DG), showed promising anticancer effects in preclinical models. 2-DG inhibits glycolysis and depletes ATP, has been applied to treat lymphoma [24] and breast cancer [25]. Therefore, based on our previous ndings and results from the literature, tumor glycolysis appears to be an ideal target for therapeutic intervention.…”

Insights into the Composition of a Co-Culture of 10 Probiotic Strains (OMNi BiOTiC® AAD10) and Effects of Its Postbiotic Culture Supernatant

“…Detected metabolites were quality controlled as described previously [ 40 , 41 ] using The Information Bus Company (TIBCO) Spotfire (v7.5.0, TIBCO, Palo Alto, CA, USA) and graded into two classes (I) suitable for multivariate and univariate analysis (MVA_UVA) and (II) suitable for univariate analysis (UVA). Technical variability was assessed calculating the relative standard deviation (RSD) of the quality control (QC) samples.…”