Tumor Biology

, Volume 37, Issue 7, pp 9967–9977 | Cite as

Downregulation of the WT1 gene expression via TMPyP4 stabilization of promoter G-quadruplexes in leukemia cells

  • Saeedeh Ghazaey Zidanloo
  • Abasalt Hosseinzadeh Colagar
  • Hossein Ayatollahi
  • Jahan-Bakhsh Raoof
Original Article

Abstract

The WT1 gene is an important oncogene, and its overexpression is considered as an effective target for anticancer therapy. Regulation of its gene transcription is one way for WT1-targeting drug design. Recently, in silico analysis of some oncogene promoters like WT1 showed some guanine-rich regions with the ability to form G-quadruplex structures. Ligands like 5,10,15,20-tetra(N-methyl-4-pyridyl)-porphine (TMPyP4) have predominant effect on G-quadruplex stabilization. The aim of this study was to understand the effect of TMPyP4 on WT1 gene transcription via stabilization of promoter G-quadruplexes. We examined the formation of new G-quadruplex motifs in WT1 promoter in the presence of TMPyP4. In order to understand the nature of its interaction with WT1 promoter quadruplexes, differential pulse voltammetry (DPV), circular dichroism (CD), polyacrylamide gel electrophoresis, electrophoretic mobility shift assay (EMSA), polymerase chain reaction (PCR) stop assays, and quantitative RT-PCR were performed. According to the results, the WT1 promoter can form stable intramolecular parallel G-quadruplexes. In addition, after 48 and 96 h of incubation, 100 μM TMPyP4 reduced the WT1 transcription to 9 and 0.4 %, respectively, compare to control. We report that ligand-mediated stabilization of G-quadruplexes within the WT1 promoter can silence WT1 expression. This study might offer the basis for the reasonable design and improvement of new porphyrin derivatives as effective anti-leukemia agents for cancer therapy.

Keywords

WT1 promoter G-quadruplex stability TMPyP4 Downregulation of transcription 

Notes

Compliance with ethical standards

Conflicts of interest

None

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Saeedeh Ghazaey Zidanloo
    • 1
  • Abasalt Hosseinzadeh Colagar
    • 1
    • 2
  • Hossein Ayatollahi
    • 3
  • Jahan-Bakhsh Raoof
    • 2
    • 4
  1. 1.Department of Molecular and Cell Biology, Faculty of Basic SciencesUniversity of MazandaranBabolsarIran
  2. 2.Nano and Biotechnology Research Group, Faculty of Basic SciencesUniversity of MazandaranBabolsarIran
  3. 3.Cancer Molecular Pathology Research CenterMashhad University of Medical SciencesMashhadIran
  4. 4.Eletroanalytical Chemistry Research Laboratory, Analytical Chemistry Department of Faculty of ChemistryMazandaran UniversityBabolsarIran

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