Aspirin induces apoptosis in mesenchymal stem cells requiring Wnt/beta-catenin pathway.
OBJECTIVE
Mesenchymal stem cells (MSC) are multipotent progenitor cells that are have found use in regenerative medicine. We have previously observed that aspirin, a widely used anti-inflammatory drug, inhibits MSC proliferation. Here we have aimed to elucidate whether aspirin induces MSC apoptosis and whether this is modulated through the Wnt/beta-catenin pathway.


METHODS
Apoptosis of MSCs was assessed using Hoechst 33342 dye and an Annexin V-FITC/PI Apoptosis Kit. Expression of protein and protein phosphorylation were investigated using Western blot analysis. Caspase-3 activity was detected by applying a caspase-3/CPP32 Colorimetric Assay Kit.


RESULTS
In these MSCs, aspirin induced morphological changes characteristic of apoptosis, cytochrome c release from mitochondria, and caspase-3 activation. Stimulating the Wnt/beta-catenin pathway by both Wnt 3a and GSK-3beta inhibitors (LiCl and SB 216763), blocked aspirin-induced apoptosis and protected mitochondrial function, as demonstrated by decreased cytochrome c release and caspase-3 activity. Aspirin initially caused a time-dependent decrease in COX-2 expression but subsequently, and unexpectedly, elevated the latter. Stimulation of COX-2 expression by aspirin was further enhanced following stimulation of the Wnt/beta-catenin pathway. Application of the COX-2 inhibitor NS-398 suppressed elevated COX-2 expression and promoted aspirin-induced apoptosis.


CONCLUSIONS
These results demonstrate that the Wnt/beta-catenin pathway is a key modulator of aspirin-induced apoptosis in MSCs by regulation of mitochrondrial/caspase-3 function. More importantly, our findings suggest that aspirin may influence MSC survival under certain conditions; therefore, it should be used with caution when considering regenerative MSC transplantation in patients with concomitant chronic inflammatory diseases such as arthritis.
Deng
L
L
Research Center for Cardiovascular Regenerative Medicine, Ministry of Health, Cardiovascular Institute & Fu Wai Hospital, Chinese Academy of Medical Science & Peking Union Medicine College, Beijing 100037, China.
Hu
S
S
Baydoun
A R
AR
Chen
J
J
Chen
X
X
Cong
X
X
D017209:Apoptosis / Q000187:drug effects*; D001241:Aspirin / Q000494:pharmacology*; D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism; D002478:Cells, Cultured; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D004789:Enzyme Activation; D005434:Flow Cytometry; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects*; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics / Q000378:metabolism; D051153:Wnt Proteins / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism*
D016861:Cyclooxygenase Inhibitors; D051153:Wnt Proteins; D051176:beta Catenin; D011451:Prostaglandin-Endoperoxide Synthases; D053148:Caspase 3; D001241:Aspirin
Curr Opin Cell Biol. 2003 Apr;15(2):158-63
12648671
Thromb Res. 2003 Jun 15;110(5-6):255-8
14592543
Biomarkers. 2007 Nov-Dec;12(6):623-34
17852081
Apoptosis. 2006 Jan;11(1):121-9
16374552
Blood. 2004 Dec 1;104(12):3581-7
15297308
N Engl J Med. 1991 Dec 5;325(23):1593-6
1669840
Circ Res. 2004 Mar 19;94(5):678-85
14739163
Expert Rev Mol Med. 2004 Oct 18;6(21):1-18
15488156
J Cell Biochem. 2004 Dec 15;93(6):1210-30
15486964
Science. 2004 Nov 26;306(5701):1568-71
15567866
Aliment Pharmacol Ther. 1997 Dec;11(6):1101-8
9663836
J Cardiovasc Pharmacol. 2004 Nov;44(5):591-5
15505497
Cell. 2006 Nov 3;127(3):469-80
17081971
J Biol Chem. 2005 Aug 5;280(31):28463-7
15946945
Mol Pharmacol. 2004 Feb;65(2):470-8
14742690
Stem Cells. 2004;22(3):377-84
15153614
Curr Opin Cell Biol. 2008 Apr;20(2):119-25
18339531
Cancer Lett. 2009 Feb 8;274(1):61-71
18849111
EMBO J. 2002 Nov 1;21(21):5635-44
12411481
Proc Natl Acad Sci U S A. 1999 May 11;96(10):5522-7
10318916
Arthritis Res Ther. 2008;10(5):223
18947375
J Pharm Pharmacol. 1971 Oct;23(10):729-44
4399908
Stem Cells. 2006 Feb;24(2):416-25
16253984
Cell. 2002 Oct 18;111(2):241-50
12408868
N Engl J Med. 2007 May 24;356(21):2131-42
17522398
Eur J Pharmacol. 2009 Jan 5;602(1):8-14
19026633
Circulation. 2000 Oct 24;102(17):2022-3
11044413
J Cell Biol. 2002 Apr 29;157(3):429-40
11980918
J Clin Invest. 1999 Mar;103(5):697-705
10074487
Oncol Rep. 2005 Apr;13(4):559-83
15756426
Nature. 2007 Oct 4;449(7162):557-63
17914389
Cell Prolif. 2006 Apr;39(2):147-55
16542349
Mol Cancer Ther. 2003 Jun;2(6):509-16
12813129
Nature. 1999 Apr 1;398(6726):422-6
10201372
J Biol Chem. 2006 Sep 29;281(39):29228-35
16880202
Circulation. 2005 Aug 23;112(8):1128-35
16103243
Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):3275-80
10077674
Proc Natl Acad Sci U S A. 2005 Nov 22;102(47):17207-12
16282376
Stem Cells. 2009 Feb;27(2):467-77
19338064
Science. 1995 Dec 22;270(5244):2017-9
8533099
Cancer Res. 2003 Feb 1;63(3):728-34
12566320
Oncogene. 2006 Oct 19;25(49):6447-56
16878161
Circulation. 2000 Mar 14;101(10):1206-18
10715270
Stem Cells. 2008 Jan;26(1):135-45
17932426
Differentiation. 2001 Oct;68(4-5):235-44
11776476
Circ Res. 2004 Jul 9;95(1):9-20
15242981
Clin Exp Pharmacol Physiol. 2006 Aug;33(8):696-701
16895542
Gene. 2009 Mar 15;433(1-2):1-7
19135507
Stem Cells. 2006 Aug;24(8):1892-903
16690780