JS-K, a nitric oxide-releasing prodrug, modulates ß-catenin/TCF signaling in leukemic Jurkat cells: evidence of an S-nitrosylated mechanism.
β-Catenin is a central player of the Wnt signaling pathway that regulates cell-cell adhesion and may promote leukemia cell proliferation. We examined whether JS-K, an NO-donating prodrug, modulates the Wnt/β-catenin/TCF-4 signaling pathway in Jurkat T-Acute Lymphoblastic Leukemia cells. JS-K inhibited Jurkat T cell growth in a concentration and time-dependent manner. The IC(50)s for cell growth inhibition were 14±0.7 and 9±1.2μM at 24 and 48h, respectively. Treatment of the cells with JS-K for 24h, caused a dose-dependent increase in apoptosis from 16±3.3% at 10μM to 74.8±2% at 100μM and a decrease in proliferation. This growth inhibition was also due, in part, to alterations in the different phases of the cell cycle. JS-K exhibited a dose-dependent cytotoxicity as measured by LDH release at 24h. However, between 2 and 8h, LDH release was less than 20% for any indicated JS-K concentration. The β-catenin/TCF-4 transcriptional inhibitory activity was reduced by 32±8, 63±5, and 93±2% at 2, 10, and 25μM JS-K, respectively, based on luciferase reporter assays. JS-K reduced nuclear β-catenin and cyclin D1 protein levels, but cytosolic β-catenin expression did not change. Based on a time-course assay of S-nitrosylation of proteins by a biotin switch assay, S-nitrsolyation of nuclear β-catenin was determined to precede its degradation. A comparison of the S-nitrosylated nuclear β-catenin to the total nuclear β-catenin showed that β-catenin protein levels were degraded at 24h, while S-nitrosylation of β-catenin occurred earlier at 0-6h. The NO scavenger PTIO abrogated the JS-K mediated degradation of β-catenin demonstrating the need for NO.
Nath
Niharika
N
Department of Physiology and Pharmacology, City University of New York Medical School, 138th Street and Convent Avenue, New York, NY 10031, United States.
Chattopadhyay
Mitali
M
Pospishil
Liliya
L
Cieciura
Lucyna Z
LZ
Goswami
Satindra
S
Kodela
Ravinder
R
Saavedra
Joseph E
JE
Keefer
Larry K
LK
Kashfi
Khosrow
K
D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D001391:Azo Compounds / Q000494:pharmacology*; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000502:physiology*; D006131:Growth Inhibitors / Q000494:pharmacology; D006801:Humans; D019169:Jurkat Cells; D009569:Nitric Oxide / Q000378:metabolism* / Q000502:physiology; D009603:Nitroso Compounds / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology*; D054218:Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D011355:Prodrugs / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D000073940:Transcription Factor 4; D014157:Transcription Factors / Q000502:physiology*; D051153:Wnt Proteins / Q000502:physiology; D051176:beta Catenin / Q000502:physiology*
D001391:Azo Compounds; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D006131:Growth Inhibitors; D009603:Nitroso Compounds; C474958:O(2)-(2,4-dinitrophenyl) 1-((4-ethoxycarbonyl)piperazin-1-yl)diazen-1-ium-1,2-diolate; D010879:Piperazines; D011355:Prodrugs; C532096:TCF4 protein, human; D000073940:Transcription Factor 4; D014157:Transcription Factors; D051153:Wnt Proteins; D051176:beta Catenin; D009569:Nitric Oxide
Blood. 1992 Oct 15;80(8):1880-4
1382708
Blood. 2007 Jul 15;110(2):709-18
17384201
Proc Natl Acad Sci U S A. 1999 May 11;96(10):5522-7
10318916
J Urol. 2004 Jul;172(1):338-44
15201807
Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):4262-6
10759547
Biochem Pharmacol. 2009 Nov 15;78(10):1298-304
19576865
Science. 2010 Mar 26;327(5973):1650-3
20339075
Leuk Res. 1995 Aug;19(8):527-33
7658698
Antioxid Redox Signal. 2006 Jul-Aug;8(7-8):1329-37
16910780
Curr Top Med Chem. 2005;5(7):625-36
16101424
Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):444-8
1346070
Bioorg Med Chem Lett. 2008 Feb 1;18(3):950-3
18178089
Nitric Oxide. 2008 Sep;19(2):177-83
18460348
Curr Top Med Chem. 2007;7(3):277-82
17305570
Biochem Biophys Res Commun. 2005 Jan 7;326(1):93-9
15567157
Leuk Res. 2006 Oct;30(10):1279-83
16439016
Nitric Oxide. 2010 May 15;22(4):259-65
19818866
Arch Biochem Biophys. 2003 Feb 15;410(2):269-79
12573287
Biochem Biophys Res Commun. 2001 Apr 20;282(5):1075-9
11302723
Blood. 2002 Aug 1;100(3):982-90
12130512
CA Cancer J Clin. 2009 Jul-Aug;59(4):225-49
19474385
Toxicol Pathol. 2007 Jun;35(4):495-516
17562483
Biochem Pharmacol. 2009 Mar 15;77(6):1064-73
19154729
Cancer Lett. 2005 Aug 8;226(1):1-15
16004928
Proc Natl Acad Sci U S A. 2003 Oct 28;100(22):12584-9
14566053
J Immunol Methods. 1988 Nov 25;115(1):61-9
3192948
Sci STKE. 2001 Jun 12;2001(86):re1
11752656
Oncogene. 2009 Jan 29;28(4):619-24
18978812
Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7674-7
1502182
Mol Cancer Ther. 2004 Jun;3(6):709-14
15210857
Clin Cancer Res. 2007 Feb 1;13(3):795-8
17289869
Nature. 1999 Apr 1;398(6726):422-6
10201372
J Org Chem. 2001 May 4;66(9):3090-8
11325274
J Cell Physiol. 2003 Dec;197(3):426-34
14566972
J Biochem. 1996 Nov;120(5):1034-9
8982873
Leuk Res. 2009 Nov;33(11):1525-9
19193435
Trends Mol Med. 2009 Sep;15(9):391-404
19726230
Blood. 1998 Jun 1;91(11):3995-4019
9596644
Proc Natl Acad Sci U S A. 2005 Nov 22;102(47):17207-12
16282376
Mol Cancer Ther. 2003 Apr;2(4):409-17
12700285
Sci STKE. 2001 Jun 12;2001(86):pl1
11752655
J Med Chem. 2006 Jul 13;49(14):4356-66
16821795
J Pharmacol Exp Ther. 2010 Nov;335(2):443-50
20679133
Bioorg Med Chem. 2008 Nov 15;16(22):9764-71
18930407