Metformin Treatment or PRODH/POX-Knock out Similarly Induces Apoptosis by Reprograming of Amino Acid Metabolism, TCA, Urea Cycle and Pentose Phosphate Pathway in MCF-7 Breast Cancer Cells.

Thi Yen Ly Huynh,Ilona Oscilowska,Jerzy Palka,Coral Barbas,Magdalena Nizioł,Jorge Sáiz,Weronika Baszanowska

doi:10.3390/biom11121888

Thi Yen Ly Huynh, Ilona Oscilowska + Show 5 more

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https://doi.org/10.3390/biom11121888

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Abstract

It has been considered that proline dehydrogenase/proline oxidase (PRODH/POX) is involved in antineoplastic activity of metformin (MET). The aim of this study is identification of key metabolites of glycolysis, pentose phosphate pathway (PPP), tricarboxylic acids (TCA), urea cycles (UC) and some amino acids in MET-treated MCF-7 cells and PRODH/POX-knocked out MCF-7 (MCF-7crPOX) cells. MCF-7crPOX cells were generated by using CRISPR-Cas9. Targeted metabolomics was performed by LC-MS/MS/QqQ. Expression of pro-apoptotic proteins was evaluated by Western blot. In the absence of glutamine, MET treatment or PRODH/POX-knock out of MCF-7 cells contributed to similar inhibition of glycolysis (drastic increase in intracellular glucose and pyruvate) and increase in the utilization of phospho-enol-pyruvic acid, glucose-6-phosphate and some metabolites of TCA and UC, contributing to apoptosis. However, in the presence of glutamine, MET treatment or PRODH/POX-knock out of MCF-7 cells contributed to utilization of some studied metabolites (except glucose), facilitating pro-survival phenotype of MCF-7 cells in these conditions. It suggests that MET treatment or PRODH/POX-knock out induce similar metabolic effects (glucose starvation) and glycolysis is tightly linked to glutamine metabolism in MCF-7 breast cancer cells. The data provide insight into mechanism of anticancer activity of MET as an approach to further studies on experimental breast cancer therapy.

Highlights

Introduction iationsBreast cancer is the most frequently diagnosed cancer in woman worldwide and has been a progressively increasing global health problem
We have suggested that MET can stimulate apoptosis in cancer cells by a cascade of processes involving the induction of AMPK, PRODH/POX and reactive oxygen species (ROS) generation under proline availability determined by several proline utilization/supporting processes
This study focused on several metabolites involved in Glycolysis, tricarboxylic acids (TCA) cycles, pentose phosphate pathway, urea cycles and several key amino acids in PRODH/POX-dependent pathways

Summary

Introduction

Breast cancer is the most frequently diagnosed cancer in woman worldwide and has been a progressively increasing global health problem. The phenotypic characteristics can be attributed to genetic and epigenetic factors, and to nonhereditary mechanisms, such as adaptive responses or fluctuations in the tumor microenvironment signaling pathways [1]. Optimal methods of treating breast cancer must be developed to effectively cure the malignancy. Metformin (MET) is currently used to treat type II diabetes patients, it evokes antineoplastic potency [2,3,4,5]. The molecular mechanism of anti-cancer activity of MET is unknown. One of the effects of MET is activation of adenosine monophosphate (AMP)

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