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PRRX1 isoforms cooperate with FOXM1 to regulate the DNA damage response in pancreatic cancer cells

Oncogene (2019) | Download Citation

Abstract

PRRX1 is a homeodomain transcriptional factor, which has two isoforms, PRXX1A and PRRX1B. The PRRX1 isoforms have been demonstrated to be important in pancreatic cancer, especially in the regulation of epithelial-to-mesenchymal transition (EMT) in Pancreatic Ductal Adenocarcinoma (PDAC) and of mesenchymal-to-epithelial transition (MET) in liver metastasis. In order to determine the functional underpinnings of PRRX1 and its isoforms, we have unraveled a new interplay between PRRX1 and the FOXM1 transcriptional factors. Our detailed biochemical analysis reveals the direct physical interaction between PRRX1 and FOXM1 proteins that requires the PRRX1A/B 200-222/217 amino acid (aa) region and the FOXM1 Forkhead domain. Additionally, we demonstrate the cooperation between PRRX1 and FOXM1 in the regulation of FOXM1-dependent transcriptional activity. Moreover, we establish FOXM1 as a critical downstream target of PRRX1 in pancreatic cancer cells. We demonstrate a novel role for PRRX1 in the regulation of genes involved in DNA repair pathways. Indeed, we show that expression of PRRX1 isoforms may limit the induction of DNA damage in pancreatic cancer cells. Finally, we demonstrate that targeting FOXM1 with the small molecule inhibitor FDI6 suppress pancreatic cancer cell proliferation and induces their apoptotic cell death. FDI6 sensitizes pancreatic cancer cells to Etoposide and Gemcitabine induced apoptosis. Our data provide new insights into PRRX1’s involvement in regulating DNA damage and provide evidence of a possible PRRX1-FOXM1 axis that is critical for PDAC cells.

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Acknowledgements

We are grateful to Dr. Zheng Fu (Virginia Commonwealth University) for providing the 6xFOXM1 luciferase reporter construct. We thank Dr. Kaori Ihida-Stansbury (University of Pennsylvania) for the PRRX1 wild type and Knockout mouse embryonic fibroblasts (MEFs). The Tenascin-C (TN7) luciferase construct was kindly provided by Dr. Edward E. Morrisey (University of Pennsylvania). We are thankful to the Molecular Pathology and Imaging Core, the Cell Culture and iPS Core and the Human-Microbial Analytic and Repository Core Facilities. This work was supported by the NIH/NIDDK R01 DK060694 (BM, MR, JRP, AKR), the Center for Molecular Studies in Digestive and Liver Diseases (NIH P30 DK050306), the American Cancer Society, Fonds de recherche en santé du Québec P-Marchand-35978 (BM), National Pancreas Foundation (MR), German Cancer Aid Foundation (Max Eder Program, Deutsche Krebshilfe 111273 to MR), AGA-Actavis Research Award in Pancreatic Disorders (MR), NCI F32 CA221094 (JRP) and the NIH loan repayment program (JRP).

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Affiliations

  1. Division of Gastroenterology, Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA

    • Benoît Marchand
    • , Jason R. Pitarresi
    • , Maximilian Reichert
    • , Kensuke Suzuki
    • , Dorottya Laczkó
    •  & Anil K. Rustgi
  2. Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA

    • Benoît Marchand
    • , Jason R. Pitarresi
    • , Kensuke Suzuki
    • , Dorottya Laczkó
    •  & Anil K. Rustgi
  3. II. Medizinische Klinik, Technical University of Munich, 81675, Munich, Germany

    • Maximilian Reichert
  4. Department of Genetics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA

    • Maximilian Reichert
    •  & Anil K. Rustgi

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The authors declare that they have no conflict of interest.

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Correspondence to Anil K. Rustgi.

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https://doi.org/10.1038/s41388-019-0725-6