2‐methoxyestradiol impacts on amino acids‐mediated metabolic reprogramming in osteosarcoma cells by its interaction with NMDA receptor

Abstract

Deregulation of serine and glycine metabolism, have been identified to function as metabolic regulators in supporting tumor cell growth. The role of serine and glycine in regulation of cancer cell proliferation is complicated, dependent on concentrations of amino acids and tissue-specific. D-serine and glycine are coagonists of N-methyl-D-aspartate (NMDA) receptor subunit GRIN1. Importantly, NMDA receptors are widely expressed in cancer cells and play an important role in regulation of cell death, proliferation, and metabolism of numerous malignancies. The aim of the present work was to associate the metabolism of glycine and D-serine with the anticancer activity of 2-methoxyestradiol. 2-methoxyestradiol is a potent anticancer agent but also a physiological 17β- estradiol metabolite. In the study we have chosen two malignant cell lines expressing functional NMDA receptors, that is osteosarcoma 143B and breast cancer MCF7. We used MTS assay, migration assay, flow cytometric analyses, Western blotting and immunoprecipitation techniques as well as molecular modeling studies. We have demonstrated the extensive crosstalk between the deregulated metabolic network and cancer cell signaling. Herein, we observed an anticancer effect of high concentrations of glycine and D-serine in osteosarcoma cells. In contrast, the amino acids when used at low, physiological concentrations induced the proliferation and migration of osteosarcoma cells. Importantly, the pro-cancergogenic effects of both glycine and D-serine where abrogated by the usage of 2-methoxyestradiol at both physiological and pharmacological relevant concentrations. The obtained data confirmed that 2-methoxyestradiol may be a physiological anticancer molecule.

2-Methoxyestradiol Reverses the Pro-Carcinogenic Effect of L-Lactate in Osteosarcoma 143B Cells

“…Importantly, the pro-migratory and proproliferative effects of L-lactate were reversed by treatment with a pharmacological concentration of 2-ME. The antimigratory potential of 2-ME has previously been demonstrated in various experimental models (17,23,24). Herein, for the first time, we demonstrated the antagonistic effect between 2-ME and L-lactate.…”

“…An anticancer mechanism of 2-ME is associated with the induction of mitochondrial oxidative stress resulting in the induction of the mitochondrial apoptotic pathway in cancer cells (38,42). It was previously demonstrated that 2-ME decreased mitochondrial membrane potential due to generation of nitro-oxidative stress leading to cancer cell death (17)(18)(19)(20)(21)43). However, 2-ME also works as an inhibitor of mitochondrial respiration by inhibiting Complex I (39-41).…”

2-Methoxyestradiol Affects Mitochondrial Biogenesis Pathway and Succinate Dehydrogenase Complex Flavoprotein Subunit A in Osteosarcoma Cancer Cells

“…Previously, we evidenced that generation of NO due to nuclear recruitment of nNOS constitutes an anticancer mechanism of 2-methoxyestradiol (2-ME) action ( Figure 1) (18). 2-ME is a natural derivative of 17β-estradiol that possesses anticancer activities, as confirmed by in vitro and in vivo studies (18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29). Under the brand name Panzem, 2-ME is being evaluated in clinical trials for the treatment of numerous malignancies, such as breast and prostate cancers (19)(20)(21)(22).…”

“…The membranes were then incubated with primary antibodies specific to α-syntrophin or PGC-1α (1:2000) overnight at 4˚C, or β-Actin (1:50000) for 30 min at room temperature. Analysis was performed as previously described (26). Chemiluminescence was detected using ImageQuant LAS 500 (GE Healthcare, Warsaw, Poland).…”

“…Under the brand name Panzem, 2-ME is being evaluated in clinical trials for the treatment of numerous malignancies, such as breast and prostate cancers (19)(20)(21)(22). Interestingly, previous studies have reported potential physiological anticancer activity of 2-ME and suggested to consider 2-ME not as a metabolite of 17β-estradiol but as a novel hormone (18,23,24,26). Nonetheless, potential hormonal properties of 2-ME need to be further elucidated by in vivo studies.…”