title: Modulation of u-PA, MMPs and their inhibitors by a novel nutrient mixture in pediatric human sarcoma cell lines.
abstract: Pediatric sarcomas are highly aggressive tumors that are characterized by high levels of matrix metalloproteinase (MMP)-2 and -9 secretions that degrade the ECM and basement membrane, allowing cancer cells to spread to distal organs. Proteases play a key role in tumor cell invasion and metastasis by digesting the basement membrane and ECM components. Strong clinical and experimental evidence demonstrates association of elevated levels of u-PA and MMPs with cancer progression, metastasis and shortened patient survival. MMP activities are regulated by specific tissue inhibitors of metalloproteinases (TIMPs). Our main objective was to study the effect of a nutrient mixture (NM) on activity of u-PA, MMPs and TIMPs in various human pediatric sarcomas. Human osteosarcoma MNNG-HOS, osteosarcoma U-2OS and rhabdomyosarcoma RD cell lines (ATCC) were cultured in their respective media and treated at confluence with NM at 0, 50, 100, 250, 500 and 1,000 µg/ml. Analysis of u-PA activity was carried out by fibrin zymography, MMPs by gelatinase zymography and TIMPs by reverse zymography. All sarcoma cell lines studied expressed u-PA, which was inhibited by NM in a dose-dependent manner. On gelatinase zymography, osteosarcoma MNNG-HOS showed a band corresponding to MMP-2 and induction of MMP-9 with PMA (100 ng/ml) treatment. U-2OS osteosarcoma cells showed strong bands corresponding to inactive MMP-2 and MMP-9 and faint bands corresponding to active MMP-2 and MMP-9 dimer; PMA treatment enhanced MMP-9 and MMP-9 dimer activity. Rhabdomyosarcoma showed MMP-2 and faint MMP-9 bands; PMA treatment enhanced MMP-9 expression. NM inhibited their expression in a dose-dependent manner. Activity of TIMPs was upregulated by NM in all cancer cell lines in a dose-dependent manner. Analysis revealed a positive correlation between u-PA and MMPs and a negative correlation between u-PA/MMPs and TIMPs. These findings suggest the therapeutic potential of NM in treatment of pediatric sarcomas.
authors.affiliation: Dr Rath Research Institute, Santa Clara, CA, USA.
authors.forename: M Waheed
authors.initials: MW
authors.lastname: Roomi
authors.forename: Tatiana
authors.initials: T
authors.lastname: Kalinovsky
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Niedzwiecki
authors.forename: Matthias
authors.initials: M
authors.lastname: Rath
mesh_terms: D001859:Bone Neoplasms / Q000178:diet therapy / Q000473:pathology; D045744:Cell Line, Tumor; D002648:Child; D005502:Food; D018093:Gelatinases; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis*; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis*; D012509:Sarcoma / Q000178:diet therapy* / Q000235:genetics / Q000473:pathology; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000096:biosynthesis*; D019716:Tissue Inhibitor of Metalloproteinase-2; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis*
chemical_list: C560822:TIMP1 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D019716:Tissue Inhibitor of Metalloproteinase-2; D014568:Urokinase-Type Plasminogen Activator; D018093:Gelatinases; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9
references.citation: Acta Orthop Scand. 2004 Aug;75(4):487-91
references.pmid: 15370596
references.citation: Annu Rev Med. 1997;48:489-509
references.pmid: 9046979
references.citation: Med Oncol. 2006;23(3):411-7
references.pmid: 17018899
references.citation: Anticancer Res. 1998 Jul-Aug;18(4A):2487-93
references.pmid: 9703897
references.citation: Med Pediatr Oncol. 1990;18(6):466-71
references.pmid: 2233517
references.citation: J Urol. 2003 Sep;170(3):773-6
references.pmid: 12913695
references.citation: Dig Dis Sci. 2003 Jan;48(1):230-7
references.pmid: 12645815
references.citation: Surg Oncol Clin N Am. 2001 Apr;10(2):383-92, x
references.pmid: 11382593
references.citation: Lab Invest. 1983 Aug;49(2):140-7
references.pmid: 6348406
references.citation: Oncol Rep. 2010 Sep;24(3):747-57
references.pmid: 20664983
references.citation: Nature. 1980 Mar 6;284(5751):67-8
references.pmid: 6243750
references.citation: Anticancer Res. 2001 Jan-Feb;21(1A):213-9
references.pmid: 11299737
references.citation: Cancer Metastasis Rev. 2010 Sep;29(3):529-42
references.pmid: 20717705
references.citation: Am J Clin Pathol. 2005 Mar;123(3):405-14
references.pmid: 15716237
references.citation: Cancer Lett. 1996 Jun 5;103(2):183-9
references.pmid: 8635156
references.citation: Proc Natl Acad Sci U S A. 2005 Sep 20;102(38):13604-9
references.pmid: 16157892
references.citation: Prostate. 1997 Aug 1;32(3):188-95
references.pmid: 9254898
references.citation: J Pediatr Hematol Oncol. 1997 Nov-Dec;19(6):483-91
references.pmid: 9407933
references.citation: Int J Cancer. 1996 Aug 22;69(4):268-72
references.pmid: 8797866
references.citation: Int J Biol Markers. 1999 Oct-Dec;14(4):268-71
references.pmid: 10669958
references.citation: Med Pediatr Oncol. 1992;20(3):209-14
references.pmid: 1574030
references.citation: Semin Oncol. 2002 Dec;29(6 Suppl 16):15-8
references.pmid: 12516034
references.citation: Med Pediatr Oncol. 1998 Dec;31(6):471-4
references.pmid: 9835898
references.citation: Cancer Metastasis Rev. 1990 Dec;9(4):289-303
references.pmid: 1965794
references.citation: Clin Orthop Relat Res. 2003 Oct;(415 Suppl):S46-58
references.pmid: 14600592
references.citation: Nutr Hosp. 1995 Nov-Dec;10(6):368-72
references.pmid: 8599623
references.citation: Cancer Chemother Pharmacol. 1999;43 Suppl:S42-51
references.pmid: 10357558
references.citation: Br J Cancer. 1982 Sep;46(3):354-67
references.pmid: 7126425
references.citation: Exp Oncol. 2010 Dec;32(4):243-8
references.pmid: 21270751
references.citation: Anticancer Drugs. 1996 Jun;7(4):461-8
references.pmid: 8826614
references.citation: J Pediatr Surg. 2004 Nov;39(11):1673-9
references.pmid: 15547833
references.citation: J Natl Cancer Inst. 1997 Sep 3;89(17):1260-70
references.pmid: 9293916
references.citation: J Clin Oncol. 2000 Mar;18(5):1135-49
references.pmid: 10694567
references.citation: Am J Clin Nutr. 2000 Jun;71(6 Suppl):1698S-702S; discussion 1703S-4S
references.pmid: 10837321
references.citation: Adv Cancer Res. 1985;44:139-266
references.pmid: 2930999
references.citation: Int J Cancer. 1997 Jul 3;72(1):1-22
references.pmid: 9212216
references.citation: FASEB J. 1990 Apr 1;4(6):1577-90
references.pmid: 2180767
references.citation: Br J Cancer. 2010 Feb 16;102(4):731-7
references.pmid: 20051950
references.citation: Nat Rev Cancer. 2002 Mar;2(3):161-74
references.pmid: 11990853
references.citation: Biochim Biophys Acta. 2002 Apr 29;1596(2):182-92
references.pmid: 12007600
references.citation: Clin Chem. 1998 Jun;44(6 Pt 1):1177-83
references.pmid: 9625040
references.citation: Histol Histopathol. 2003 Oct;18(4):1035-40
references.pmid: 12973672
references.citation: Int J Urol. 2003 Mar;10(3):160-6
references.pmid: 12622713
references.citation: Cancer. 1999 Oct 15;86(8):1602-8
references.pmid: 10526292
references.citation: J Biol Chem. 2001 Jun 8;276(23):20085-92
references.pmid: 11274215
references.citation: Cancer Lett. 1992 Jul 31;65(1):51-4
references.pmid: 1511409
references.citation: Carcinogenesis. 1998 Apr;19(4):611-6
references.pmid: 9600345
references.citation: J Clin Oncol. 2009 Jun 1;27(16):2712-25
references.pmid: 19414669
references.citation: Mod Pathol. 2001 May;14(5):506-14
references.pmid: 11353062
references.citation: J Surg Res. 2005 Aug;127(2):151-6
references.pmid: 16083752