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Metformin mediated reversal of epithelial to mesenchymal transition is triggered by epigenetic changes in E-cadherin promoter

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Abstract

Epithelial-mesenchymal transition (EMT) is one of the key biological phenomena behind cancer and metastasis. Clinical studies suggest that patients undergoing metformin therapy are less predisposed to cancer but the underlying mechanism is far from clear. Given that metformin also acts as TGF-β inhibitor, we sought to explore whether and how metformin could modulate EMT in a cancer like microenvironment. Our data using human cell lines revealed that metformin induced a distinct change from stromal-shaped mesenchymal cells to cuboidal-shaped epithelial cells with upregulation of epithelial markers and mitigation of their invasive property. One of the key regulatory pathways, which intersect tumorigenesis and metformin activity, is AMPK. We demonstrated that metformin attenuates ERK signaling by activating AMPK pathway leading to suppression of Snail and Slug resulting in upregulation of crucial tumor suppressor gene E-cadherin. ChIP assay confirmed insufficient binding of repressors like Slug to the E-cadherin promoter. Further, our data revealed reduction in HDAC activity prompting hypomethylation of E-cadherin promoter thus reflecting an epigenetic modification. To expand the translational significance of the study we verified these findings in diabetic patients undergoing metformin treatment. To our knowledge this is the first report representing an inverse relationship of AMPK and ERK signaling axis in promoting mesenchymal to epithelial transition (MET) via re-expression of E-cadherin upon metformin treatment thus rationalizing lower incidence of cancer in metformin-administered patients.

Key message

  • Metformin promotes reversal of the epithelial-mesenchymal transition.

  • Metformin attenuates ERK signaling by activating AMP kinase.

  • Metformin induces hypomethylation of the E-cadherin gene promoter.

  • Epigenetic modification of the E-cadherin promoter was observed in leukocytes from diabetic subjects.

  • These findings provide a potential basis for decreased cancer incidence in metformin-treated subjects.

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Acknowledgments

The authors would like to thank Dr. Antonio Garcia de Herreros, IMIM-Hospital del Mar, Barcelona, Spain; Dr. James Nelson, Stanford University, CA, USA and Dr. K. Satyamoorthy, Manipal University, Manipal, India for providing the plasmids. We gratefully thank the support extended by Manipal University, Manipal for the required facilities and intramural funding for carrying out the present study.

Author’s contribution

Poulomi Banerjee (P.B.): design, collection and assembly of data, data analysis and interpretation, manuscript writing; Harshini Surendran (H.S.): collection of data, analysis and interpretation; Debabani Roy Chowdhury (D.R.C.): collection of data, analysis and interpretation; Karthik Prabhakar (K.P.): recruitment of patients, blood sample collection and final approval of manuscript; Rajarshi Pal (R.P.): Conception and design, data analysis and interpretation, final approval of the manuscript, financial support.

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Authors and Affiliations

  1. School of Regenerative Medicine, Manipal University, Bangalore, 560 065, India

    Poulomi Banerjee, Harshini Surendran & Rajarshi Pal

  2. Department of Diabetes and Endocrinology, Manipal Hospital, Bangalore, 560 017, India

    Karthik Prabhakar

  3. Functional Genomics and Gene Silencing Group, Centre for Cellular and Molecular Biology, Hyderabad, 500 007, India

    Debabani Roy Chowdhury

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  1. Poulomi Banerjee
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Correspondence to Rajarshi Pal.

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Banerjee, P., Surendran, H., Chowdhury, D.R. et al. Metformin mediated reversal of epithelial to mesenchymal transition is triggered by epigenetic changes in E-cadherin promoter. J Mol Med 94, 1397–1409 (2016). https://doi.org/10.1007/s00109-016-1455-7

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  • DOI: https://doi.org/10.1007/s00109-016-1455-7

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