title: Downregulation of the WT1 gene expression via TMPyP4 stabilization of promoter G-quadruplexes in leukemia cells.
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.
authors.affiliation: Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, CP: 47416-95447, Iran.
authors.forename: Saeedeh Ghazaey
authors.initials: SG
authors.lastname: Zidanloo
authors.affiliation: Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, CP: 47416-95447, Iran. ahcolagar@umz.ac.ir.|Nano and Biotechnology Research Group, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Iran. ahcolagar@umz.ac.ir.
authors.forename: Abasalt
authors.initials: A
authors.lastname: Hosseinzadeh Colagar
authors.affiliation: Cancer Molecular Pathology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Ayatollahi
authors.affiliation: Nano and Biotechnology Research Group, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Iran.|Eletroanalytical Chemistry Research Laboratory, Analytical Chemistry Department of Faculty of Chemistry, Mazandaran University, Babolsar, Iran.
authors.forename: Jahan-Bakhsh
authors.initials: JB
authors.lastname: Raoof
mesh_terms: D049109:Cell Proliferation; D002942:Circular Dichroism; D004273:DNA, Neoplasm / Q000737:chemistry*; D024202:Electrophoretic Mobility Shift Assay; D054856:G-Quadruplexes; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D008024:Ligands; D008958:Models, Molecular; D011166:Porphyrins / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D025721:WT1 Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism
chemical_list: D004273:DNA, Neoplasm; D008024:Ligands; D011166:Porphyrins; D012333:RNA, Messenger; D025721:WT1 Proteins; C583751:WT1 protein, human; C021096:tetra(4-N-methylpyridyl)porphine
keywords: Downregulation of transcription; G-quadruplex stability; TMPyP4; WT1 promoter
references.citation: Nucleic Acids Res. 2007;35(22):7698-713
references.pmid: 17984069
references.citation: Haematologica. 2007 Sep;92(9):1270-2
references.pmid: 17768125
references.citation: Melanoma Res. 2007 Dec;17(6):341-8
references.pmid: 17992115
references.citation: Future Med Chem. 2010 Apr;2(4):619-46
references.pmid: 20563318
references.citation: Nucleic Acids Res. 2006 Jul 1;34(Web Server issue):W676-82
references.pmid: 16845096
references.citation: Leukemia. 2005 Aug;19(8):1416-23
references.pmid: 15920493
references.citation: Nucleic Acids Res. 2008 Jan;36(Database issue):D372-6
references.pmid: 17916572
references.citation: J Neurosci. 2000 Jan 15;20(2):600-5
references.pmid: 10632589
references.citation: Cancer Res. 1988 Feb 1;48(3):589-601
references.pmid: 3335022
references.citation: J Am Chem Soc. 2005 Aug 3;127(30):10584-9
references.pmid: 16045346
references.citation: Phys Chem Chem Phys. 2013 Jun 21;15(23):9117-24
references.pmid: 23644414
references.citation: Genes Dev. 1988 Dec;2(12B):1700-12
references.pmid: 3071490
references.citation: J Neurooncol. 2007 Jun;83(2):163-72
references.pmid: 17206472
references.citation: Biochemistry. 1999 Dec 7;38(49):16067-75
references.pmid: 10587429
references.citation: Biochem Biophys Res Commun. 2002 Jul 26;295(4):784-90
references.pmid: 12127961
references.citation: Leukemia. 2007 Oct;21(10 ):2164-70
references.pmid: 17690705
references.citation: Biochem Soc Trans. 2001 Nov;29(Pt 6):692-6
references.pmid: 11709056
references.citation: Zhonghua Xue Ye Xue Za Zhi. 2002 Jul;23(7):356-9
references.pmid: 12411052
references.citation: Biophys J. 2010 Jun 2;98(11):2628-33
references.pmid: 20513407
references.citation: Proc Natl Acad Sci U S A. 2002 Sep 3;99(18):11593-8
references.pmid: 12195017
references.citation: Rep Biochem Mol Biol. 2014 Oct;3(1):7-13
references.pmid: 26989731
references.citation: Pflugers Arch. 2008 Feb;455(5):839-47
references.pmid: 17912546
references.citation: Biochimie. 2009 Jul;91(7):894-902
references.pmid: 19401211
references.citation: Biochemistry. 2005 Dec 13;44(49):16341-50
references.pmid: 16331995
references.citation: Nucleic Acids Res. 2007;35(22):7429-55
references.pmid: 17913750
references.citation: Nucleic Acids Res. 2003 Feb 15;31(4):1156-63
references.pmid: 12582234
references.citation: Biochimie. 2008 Aug;90(8):1140-8
references.pmid: 18294969
references.citation: Mol Cancer Ther. 2008 Apr;7(4):880-9
references.pmid: 18413801
references.citation: J Am Chem Soc. 2001 Sep 19;123(37):8902-13
references.pmid: 11552797
references.citation: Biochim Biophys Acta. 2012 Dec;1820(12):1987-96
references.pmid: 23000573
references.citation: Nucleic Acids Res. 2009 Sep;37(17):5749-56
references.pmid: 19617376
references.citation: Nucleic Acids Res. 2006 May 10;34(9):2536-49
references.pmid: 16687659
references.citation: Nat Chem Biol. 2007 Apr;3(4):218-21
references.pmid: 17322877
references.citation: Bioorg Med Chem Lett. 2012 Jan 1;22(1):518-22
references.pmid: 22104143
references.citation: Biochem Biophys Res Commun. 2004 Oct 22;323(3):802-8
references.pmid: 15381071
references.citation: Nucleic Acids Res. 2002 May 1;30(9):e36
references.pmid: 11972351
references.citation: Nature. 1993 Jun 10;363(6429):561-5
references.pmid: 8389423
references.citation: Biochimie. 2008 Aug;90(8):1149-71
references.pmid: 18355457
references.citation: Leukemia. 2002 Oct;16(10):2115-21
references.pmid: 12357365
references.citation: Biochemistry. 2006 Jun 27;45(25):7854-60
references.pmid: 16784237
references.citation: Nucleic Acids Res. 2005 Oct 20;33(18):6070-80
references.pmid: 16239639
references.citation: Blood. 1994 Nov 1;84(9):3071-9
references.pmid: 7949179
references.citation: Mol Cancer Ther. 2002 Jun;1(8):565-73
references.pmid: 12479216
references.citation: J Med Assoc Thai. 2007 Nov;90(11):2416-21
references.pmid: 18181329
references.citation: Proc Natl Acad Sci U S A. 1986 Oct;83(19):7241-5
references.pmid: 3463962
references.citation: Leukemia. 2003 Jul;17(7):1301-12
references.pmid: 12835718
references.citation: J Biol Chem. 1994 Mar 25;269(12):8892-900
references.pmid: 8132626