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

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Tumor Biology

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.

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Authors and Affiliations

  1. Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, CP: 47416-95447, Iran

    Saeedeh Ghazaey Zidanloo & Abasalt Hosseinzadeh Colagar

  2. Nano and Biotechnology Research Group, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Iran

    Abasalt Hosseinzadeh Colagar & Jahan-Bakhsh Raoof

  3. Cancer Molecular Pathology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Hossein Ayatollahi

  4. Eletroanalytical Chemistry Research Laboratory, Analytical Chemistry Department of Faculty of Chemistry, Mazandaran University, Babolsar, Iran

    Jahan-Bakhsh Raoof

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  1. Saeedeh Ghazaey Zidanloo
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Zidanloo, S.G., Hosseinzadeh Colagar, A., Ayatollahi, H. et al. Downregulation of the WT1 gene expression via TMPyP4 stabilization of promoter G-quadruplexes in leukemia cells. Tumor Biol. 37, 9967–9977 (2016). https://doi.org/10.1007/s13277-016-4881-9

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