Abstract
TWIST1 is a major regulator of epithelial mesenchymal transition process, essential in cancer metastasis. Cancer cells increase glucose uptake capabilities to meet their high energy requirements. In this study, we explored the potential role of TWIST1 on glucose transport into the 293T cells in an insulin-dependent and insulin-independent manner. For this purpose, the ectopic expression of TWIST1 was successfully performed by electroporation. The altered mRNA expressions of GLUT-1, -3, -4, and -12, insulin receptor (InsR), and insulin receptor substrate (IRS)-1 and -2 were assessed in control and TWIST1-overexpressing cells. Glucose uptake rates of the cells were evaluated by fluorometric glucose uptake assay. Our findings showed that the transcriptional expression levels of GLUT-1, -3, and -12 genes were significantly upregulated by TWIST1. However, TWIST1 did not alter the mRNA and protein expressions of the InsR, its substrates (IRS-1 and -2), and GLUT-4 genes in 293T cells which are main factors for insulin-stimulated glucose uptake pathway. Also, the glucose transport activities were significantly increased in TWIST1-overexpressing cells compared to controls due to fetal bovine serum (FBS) stimulation, but there was a slight non-significant difference in insulin stimulation. Thus, our data suggest that TWIST1 could promote glucose uptake independently of insulin and is possible to be evaluated as a metabolic marker in cancer. Further investigations are needed to clarify the precise molecular mechanisms underlying the cells’ glucose uptake and consumption during tumorigenesis.
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Acknowledgements
The authors are thankful for contributions to Gerard Wagemaker from Erasmus University Rotterdam, Duygu Uckan Cetinkaya, and Fatima Susanna F. Aerts Kaya from Hacettepe University, and Osman Nidai Ozes from Akdeniz University.
Funding
This work was supported by Scientific Research Projects Coordination Unit of Necmettin Erbakan University (Project no: 161315002).
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S. Pehlivanoglu: designed the experiments, performed cell culture and western blot assay, analysis of obtained data and drafted the manuscript, final approval of the version to be published. O.B. Sahan: performed electroporation method and carried out immunofluorescent analysis. S. Pehlivanoglu: carried out qRT-PCR analysis and contributed to the writing of the manuscript. K. Aktas Kont: contributed to the cell culture and qRT-PCR analysis. The manuscript has been reviewed by S. Pehlivanoglu.
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Pehlivanoglu, S., Sahan, O.B., Pehlivanoglu, S. et al. Epithelial mesenchymal transition regulator TWIST1 transcription factor stimulates glucose uptake through upregulation of GLUT1, GLUT3, and GLUT12 in vitro. In Vitro Cell.Dev.Biol.-Animal 57, 933–943 (2021). https://doi.org/10.1007/s11626-021-00635-w
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DOI: https://doi.org/10.1007/s11626-021-00635-w