Citation

  • Authors: Zizza, P., Dinami, R., Porru, M., Cingolani, C., Salvati, E., Rizzo, A., D'Angelo, C., Petti, E., Amoreo, C. A., Mottolese, M., Sperduti, I., Chambery, A., Russo, R., Ostano, P., Chiorino, G., Blandino, G., Sacconi, A., Cherfils-Vicini, J., Leonetti, C., Gilson, E., Biroccio, A.
  • Year: 2019
  • Journal: Nucleic Acids Res 47 3365-3382
  • Applications: in vitro / DNA, shRNA and DNA cotransfection, siRNA / INTERFERin, jetPEI
  • Cell types:
    1. Name: HCT 116
      Description: Human colon carcinoma cells
      Known as: HCT116
    2. Name: Phoenix
      Description: Human embryonic kidney Fibroblast transformed with adenovirus E1a. Available through Orbigen.

Method

Amphotropic Retrovirus production in Phoenix cells

Abstract

The telomeric protein TRF2 is overexpressed in several human malignancies and contributes to tumorigenesis even though the molecular mechanism is not completely understood. By using a high-throughput approach based on the multiplexed Luminex X-MAP technology, we demonstrated that TRF2 dramatically affects VEGF-A level in the secretome of cancer cells, promoting endothelial cell-differentiation and angiogenesis. The pro-angiogenic effect of TRF2 is independent from its role in telomere capping. Instead, TRF2 binding to a distal regulatory element promotes the expression of SULF2, an endoglucosamine-6-sulfatase that impairs the VEGF-A association to the plasma membrane by inducing post-synthetic modification of heparan sulfate proteoglycans (HSPGs). Finally, we addressed the clinical relevance of our findings showing that TRF2/SULF2 expression is a worse prognostic biomarker in colorectal cancer (CRC) patients.

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