Characterization of the anti-angiogenic properties of arresten, an alpha1beta1 integrin-dependent collagen-derived tumor suppressor.
Physiological and pathological turnover of basement membranes liberates biologically active cryptic molecules. Several collagen-derived fragments possess anti-angiogenic activity. Arresten is the 26-kDa non-collagenous domain of type IV collagen alpha1 chain. It functions as an efficient inhibitor of angiogenesis and tumor growth in mouse models, but its anti-angiogenic mechanism is not completely known. Here we show that arresten significantly increases apoptosis of endothelial cells in vitro by decreasing the amount of anti-apoptotic molecules of the Bcl-family; Bcl-2 and Bcl-xL. Although the pro-apoptotic effect of arresten is endothelial cell specific in vitro, in mouse tumors arresten induced apoptosis both in endothelial and tumor cells. The tumor cell apoptosis is likely an indirect effect due to the inhibition of blood vessel growth into the tumor. The active site of arresten was localized by deletion mutagenesis within the C-terminal half of the molecule. We have previously shown that arresten binds to alpha1beta1 integrin on human umbilical vein endothelial cells. However, the microvascular endothelial cells (MLECs) are more important in the context of tumor vasculature. We show here that arresten binds also to the microvascular endothelial cells via alpha1beta1 integrin. Furthermore, it has no effect on Matrigel neovascularization or the viability of integrin alpha1 null MLECs. Tumors implanted on integrin alpha1 deficient mice show no integrin alpha1 expression in the host-derived vascular endothelium, and thus arresten does not inhibit the tumor growth. Collectively, this data sheds more light into the anti-angiogenic mechanism of arresten.
Nyberg
Pia
P
Center for Matrix Biology, Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA, USA. pia.nyberg@oulu.fi
Xie
Liang
L
Sugimoto
Hikaru
H
Colorado
Pablo
P
Sund
Malin
M
Holthaus
Kathryn
K
Sudhakar
Akulapalli
A
Salo
Tuula
T
Kalluri
Raghu
R
D020533:Angiogenesis Inhibitors / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D001011:Aorta / Q000166:cytology; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002417:Cattle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000737:chemistry* / Q000235:genetics; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D039222:Integrin alpha1beta1 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary / Q000502:physiology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000187:drug effects / Q000378:metabolism; D011651:Pulmonary Artery / Q000166:cytology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D051020:bcl-X Protein / Q000187:drug effects / Q000378:metabolism
D020533:Angiogenesis Inhibitors; D024141:Collagen Type IV; D039222:Integrin alpha1beta1; D019253:Proto-Oncogene Proteins c-bcl-2; D011994:Recombinant Proteins; D025521:Tumor Suppressor Proteins; D051020:bcl-X Protein
Crit Rev Biochem Mol Biol. 1992;27(1-2):93-127
1309319
Mol Cell. 2001 Apr;7(4):811-22
11336704
Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):13612-7
9391074
Biopolymers. 2004;76(1):34-47
14997473
J Biol Chem. 2001 Aug 24;276(34):31959-68
11399763
Oncogene. 2002 Jan 10;21(2):272-81
11803470
Cancer Cell. 2003 Jun;3(6):589-601
12842087
Biochem Biophys Res Commun. 2004 May 28;318(2):354-60
15120609
Lab Invest. 1992 Oct;67(4):519-28
1279270
Nat Med. 1995 Jan;1(1):27-31
7584949
Cancer Res. 1998 Sep 1;58(17):3918-25
9731503
Transpl Immunol. 1997 Sep;5(3):179-83
9402683
Clin Biochem. 1997 Aug;30(6):455-63
9316739
J Cell Biol. 2004 Jul 5;166(1):97-109
15240572
J Cell Biol. 1998 Jul 27;142(2):587-94
9679154
Cardiovasc Res. 2004 Sep 1;63(4):603-10
15306215
J Cell Biol. 2001 Sep 3;154(5):1069-79
11535623
Dev Biol. 1996 May 1;175(2):301-13
8626034
Proc Natl Acad Sci U S A. 2000 Feb 29;97(5):2202-7
10681423
Cell. 1997 Jan 24;88(2):277-85
9008168
J Biol Chem. 2000 Jul 21;275(29):21785-8
10801897
Lab Invest. 1995 Mar;72(3):367-75
7898055
Oncogene. 2003 Nov 24;22(53):8590-607
14634621
Semin Cancer Biol. 2003 Apr;13(2):159-67
12654259
Annu Rev Biochem. 1995;64:403-34
7574488
Proc Natl Acad Sci U S A. 2003 Apr 15;100(8):4766-71
12682293
Nat Rev Cancer. 2002 Oct;2(10):727-39
12360276
Clin Cancer Res. 2002 Jul;8(7):2413-22
12114447
Proc Natl Acad Sci U S A. 2001 Jan 30;98(3):1024-9
11158588
J Biol Chem. 2000 Jan 14;275(2):1209-15
10625665
J Biol Chem. 1998 Dec 11;273(50):33287-94
9837901
J Biol Chem. 1991 Jun 25;266(18):11436-41
1646806
J Biol Chem. 2000 Jan 7;275(1):35-40
10617582
Cancer Res. 2000 May 1;60(9):2520-6
10811134
J Biol Chem. 2003 Jun 20;278(25):22404-11
12690120
J Biol Chem. 2000 Aug 4;275(31):23745-50
10837460
Biochem Biophys Res Commun. 2003 Dec 19;312(3):801-5
14680836
J Biol Chem. 2000 Mar 17;275(11):8051-61
10713126
J Biol Chem. 2003 Sep 26;278(39):37632-6
12876280
J Biol Chem. 2000 Jul 14;275(28):21340-8
10766752
J Cell Biol. 1991 Aug;114(4):855-63
1869591
Cancer Res. 1999 Sep 15;59(18):4584-90
10493512
J Clin Invest. 2005 Oct;115(10):2801-10
16151532
Nat Rev Cancer. 2003 Jun;3(6):422-33
12778132
Microvasc Res. 2007 Sep-Nov;74(2-3):85-9
17602710