ISSN 1301 - 0883 / E-ISSN 1309 - 3886

  TWIST1-overexpression induces glycolytic reprogramming and ATP production in 293T cells [Eastern J Med]
Eastern J Med. 2020; 25(3): 415-421 | DOI: 10.5505/ejm.2020.63496  

TWIST1-overexpression induces glycolytic reprogramming and ATP production in 293T cells

Suray Pehlivanoğlu, Kadriye Aktaş Kont
Necmettin Erbakan University, Faculty of Science, Department of Molecular Biology and Genetics.

INTRODUCTION: Reprogramming of glucose metabolism in cancer cells triggers tumor development. Most cancer cells provide the energy needed from aerobic glycolysis called the "Warburg effect" to promote uncontrolled proliferation and invasion. Therefore, direct regulator molecules of aerobic glycolysis remain active research targets. Epithelial mesenchymal transition (EMT) is the main mechanism that promotes cancer invasion and poor prognosis. One of the key effector molecules of the EMT is TWIST1. In this study, we sought to investigate the role that TWIST1 has in driving glycolytic programming and cellular energy charges in 293T cells by overexpressing TWIST1 in 293T cells.
METHODS: Plasmid vectors were successfully transfected by lipofectamine 2000 and mRNA expressions of interested genes were assessed. Glucose, lactate and pH levels of culture supernatatnts were determined by radiometer analyzer. The cellular energy charge of the cells were calculated from ATP, ADP and AMP data analyzed by HPLC.
RESULTS: Here, we found that TWIST1 transcription factor, which has highly conserved sequences, is an important regulator for aerobic glycolysis. We found that TWIST1 increases the expression of glycolytic genes such as HK2, PKM2, LDHα, PFKM and G6PD, the production of lactate and extracellular acidification in 293T cells. We have also determined that TWIST1 promotes aerobic glycolysis metabolism by providing cellular energy exchange. TWIST1 overexpression reduced AMP/ATP and ADP/ATP ratios in 293T cells, with further increase seen in ATP production.
DISCUSSION AND CONCLUSION: By this work, we confirmed that TWIST1 is closely related to the glycolysis pathway and is an important regulator of the Warburg effect.

Keywords: TWIST1, Glycolysis, ATP.

Suray Pehlivanoğlu, Kadriye Aktaş Kont. TWIST1-overexpression induces glycolytic reprogramming and ATP production in 293T cells. Eastern J Med. 2020; 25(3): 415-421

Corresponding Author: Suray Pehlivanoğlu, Türkiye

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