title: Transforming growth factor beta3 regulates the versican variants in the extracellular matrix-rich uterine leiomyomas.
abstract: Uterine leiomyoma are common, benign tumors that are enriched in extracellular matrix. The tumors are characterized by a disoriented and loosely packed collagen fibril structure similar to other diseases with disrupted Transforming growth factor beta (TGF-beta) signaling. Here we characterized TGF-beta3 signaling and the expression patterns of the critical extracellular matrix component versican in leiomyoma and myometrial tissue and cell culture. We also demonstrate the regulation of the versican variants by TGF-beta3. Using leiomyoma and matched myometrium from 15 patients, messenger RNA (mRNA) from leiomyoma and myometrium was analyzed by semiquantitative real time reverse transcription-polymerase chain reaction (RT-PCR), while protein analysis was done by western blot. Transforming growth factor beta3 transcripts were increased 4-fold in leiomyoma versus matched myometrium. Phosphorylated-TGF-beta RII and phosphorylated-Smad 2/3 complex were greater in leiomyoma as documented by Western blot. The inhibitor Smad7 transcripts were decreased 0.44-fold. The glycosaminoglycan (GAG)-rich versican variants were elevated in leiomyoma versus myometrial tissue: specifically V0 (4.27 +/- 1.12) and V1 (2.01 +/- 0.27). Treatment of leiomyoma and myometrial cells with TGF-beta3 increased GAG-rich versican variant expression 7 to 12 fold. Neutralizing TGF-beta3 antibody decreased the expression of the GAG-rich versican variants 2 to 8 fold in leiomyoma cells. Taken together, the aberrant production of excessive and disorganized extracellular matrix that defines the leiomyoma phenotype involves the activation of the TGF-beta signaling pathway and excessive production of GAG-rich versican variants.
authors.affiliation: Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.
authors.forename: John M
authors.initials: JM
authors.lastname: Norian
authors.forename: Minnie
authors.initials: M
authors.lastname: Malik
authors.forename: Candace Y
authors.initials: CY
authors.lastname: Parker
authors.forename: Doina
authors.initials: D
authors.lastname: Joseph
authors.forename: Phyllis C
authors.initials: PC
authors.lastname: Leppert
authors.forename: James H
authors.initials: JH
authors.lastname: Segars
authors.forename: William H
authors.initials: WH
authors.lastname: Catherino
mesh_terms: D015153:Blotting, Western; D016022:Case-Control Studies; D002460:Cell Line; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D007889:Leiomyoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D010766:Phosphorylation; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D051899:Smad2 Protein / Q000378:metabolism; D051900:Smad3 Protein / Q000378:metabolism; D051906:Smad7 Protein / Q000378:metabolism; D053782:Transforming Growth Factor beta3 / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation; D014594:Uterine Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D053675:Versicans / Q000378:metabolism*
chemical_list: D006025:Glycosaminoglycans; D012333:RNA, Messenger; C497782:SMAD2 protein, human; C497790:SMAD3 protein, human; C497806:SMAD7 protein, human; D051899:Smad2 Protein; D051900:Smad3 Protein; D051906:Smad7 Protein; D053782:Transforming Growth Factor beta3; C508322:VCAN protein, human; D053675:Versicans
references.citation: Biochim Biophys Acta. 2003 Jan 2;1619(1):98-112
references.pmid: 12495820
references.citation: Annu Rev Cell Biol. 1990;6:597-641
references.pmid: 2177343
references.citation: Circ Res. 2004 May 14;94(9):1158-67
references.pmid: 15142969
references.citation: J Biol Chem. 2002 Feb 8;277(6):4565-72
references.pmid: 11726670
references.citation: J Biol Chem. 1998 Jan 2;273(1):338-43
references.pmid: 9417085
references.citation: J Biol Chem. 1992 May 15;267(14):10003-10
references.pmid: 1577773
references.citation: Fertil Steril. 2002 Jul;78(1):1-12
references.pmid: 12095482
references.citation: Endocr Relat Cancer. 2006 Jun;13(2):379-400
references.pmid: 16728569
references.citation: Obstet Gynecol Clin North Am. 2006 Mar;33(1):41-58
references.pmid: 16504805
references.citation: EMBO J. 1989 Oct;8(10):2975-81
references.pmid: 2583089
references.citation: Fertil Steril. 2003 Aug;80(2):282-90
references.pmid: 12909487
references.citation: Proc Natl Acad Sci U S A. 1997 Sep 16;94(19):10116-21
references.pmid: 9294172
references.citation: Circ Res. 2006 Feb 17;98(3):370-7
references.pmid: 16385080
references.citation: Cell Res. 2005 Jul;15(7):483-94
references.pmid: 16045811
references.citation: J Biol Chem. 1999 Jul 16;274(29):20444-9
references.pmid: 10400671
references.citation: Methods Mol Med. 2003;81:69-77
references.pmid: 12725115
references.citation: J Immunol. 2005 Oct 15;175(8):5390-5
references.pmid: 16210645
references.citation: J Clin Endocrinol Metab. 2003 Mar;88(3):1350-61
references.pmid: 12629129
references.citation: Obstet Gynecol. 1998 Oct;92(4 Pt 1):624-7
references.pmid: 9764641
references.citation: Biochimie. 2001 Jun;83(6):529-36
references.pmid: 11506898
references.citation: Cancer Res. 1999 May 15;59(10):2324-8
references.pmid: 10344737
references.citation: Gynecol Obstet Invest. 1995;40(2):120-4
references.pmid: 8575689
references.citation: J Clin Endocrinol Metab. 1994 Sep;79(3):900-6
references.pmid: 8077380
references.citation: EMBO J. 1988 Dec 1;7(12):3737-43
references.pmid: 3208746
references.citation: Fertil Steril. 2000 May;73(5):1006-11
references.pmid: 10785229
references.citation: Rheumatology (Oxford). 2004 Aug;43(8):969-72
references.pmid: 15138331
references.citation: Am J Physiol Lung Cell Mol Physiol. 2007 Sep;293(3):L525-34
references.pmid: 17631612
references.citation: Cell. 2006 Jun 2;125(5):929-41
references.pmid: 16751102
references.citation: Mol Cell Endocrinol. 2003 Nov 14;209(1-2):9-16
references.pmid: 14604812
references.citation: Fertil Steril. 2004 Oct;82 Suppl 3:1182-7
references.pmid: 15474093
references.citation: J Cell Physiol. 2002 Jan;190(1):38-45
references.pmid: 11807809
references.citation: EMBO J. 2000 Apr 17;19(8):1745-54
references.pmid: 10775259
references.citation: Nat Rev Cancer. 2006 Jul;6(7):506-20
references.pmid: 16794634
references.citation: Mol Biol Cell. 2004 May;15(5):2093-104
references.pmid: 14978219
references.citation: FASEB J. 2004 Apr;18(6):754-6
references.pmid: 14766798
references.citation: J Clin Endocrinol Metab. 2001 Feb;86(2):913-20
references.pmid: 11158066
references.citation: Autoimmun Rev. 2006 Oct;5(8):563-9
references.pmid: 17027893
references.citation: Genes Chromosomes Cancer. 2004 Jul;40(3):204-17
references.pmid: 15139000
references.citation: Endocrinology. 2005 Mar;146(3):1097-118
references.pmid: 15604209
references.citation: Fertil Steril. 2002 Jul;78(1):114-21
references.pmid: 12095500
references.citation: Hepatology. 2002 May;35(5):1022-30
references.pmid: 11981752
references.citation: Kidney Int. 2006 Jan;69(2):213-7
references.pmid: 16408108
references.citation: Mol Cell Biochem. 1998 Dec;189(1-2):145-52
references.pmid: 9879665
references.citation: Nucleic Acids Res. 2001 May 1;29(9):e45
references.pmid: 11328886
references.citation: J Biol Chem. 1986 Oct 15;261(29):13526-35
references.pmid: 3759976
references.citation: J Biol Chem. 2006 Jan 27;281(4):2390-400
references.pmid: 16257955
references.citation: J Biol Chem. 2000 Nov 10;275(45):35448-56
references.pmid: 10950950
references.citation: J Cell Biol. 2004 Sep 13;166(6):877-87
references.pmid: 15364962
references.citation: Am J Obstet Gynecol. 2008 Apr;198(4):474.e1-11
references.pmid: 18395046
references.citation: Biochim Biophys Acta. 1998 Mar 5;1406(2):203-13
references.pmid: 9573366
references.citation: Mol Hum Reprod. 2006 Apr;12(4):245-56
references.pmid: 16571622
references.citation: Arch Biochem Biophys. 2001 Oct 1;394(1):29-38
references.pmid: 11566024
references.citation: Curr Opin Cell Biol. 2002 Oct;14(5):617-23
references.pmid: 12231358
references.citation: Hum Reprod. 2005 Apr;20(4):852-63
references.pmid: 15705628
references.citation: Fertil Steril. 2007 May;87(5):1166-72
references.pmid: 17222838
references.citation: J Biol Chem. 2002 Dec 6;277(49):47626-35
references.pmid: 12221092
references.citation: J Biol Chem. 1994 Dec 30;269(52):32992-8
references.pmid: 7806529
references.citation: Mol Hum Reprod. 2005 Jul;11(7):489-94
references.pmid: 16123076
references.citation: Cancer Cell. 2005 Sep;8(3):241-54
references.pmid: 16169468
references.citation: Am J Obstet Gynecol. 2006 Aug;195(2):415-20
references.pmid: 16635466
references.citation: J Biol Chem. 2002 Apr 12;277(15):12921-30
references.pmid: 11821431
references.citation: J Cell Sci. 2001 Dec;114(Pt 24):4359-69
references.pmid: 11792802
references.citation: N Engl J Med. 2000 May 4;342(18):1350-8
references.pmid: 10793168
references.citation: Clin Endocrinol (Oxf). 2008 Sep;69(3):462-70
references.pmid: 18248652
references.citation: Clin Cancer Res. 2002 Apr;8(4):1054-60
references.pmid: 11948113
references.citation: J Biol Chem. 2001 Jan 12;276(2):1253-61
references.pmid: 11038354
references.citation: Differentiation. 2003 Apr;71(3):171-205
references.pmid: 12694202
references.citation: Trends Biochem Sci. 2004 May;29(5):265-73
references.pmid: 15130563
references.citation: J Biol Chem. 2002 Apr 5;277(14):12294-301
references.pmid: 11805102
references.citation: Mol Hum Reprod. 1997 Mar;3(3):233-40
references.pmid: 9237249
references.citation: J Biol Chem. 1994 Dec 30;269(52):32999-3008
references.pmid: 7528742
references.citation: Radiology. 1989 Apr;171(1):241-3
references.pmid: 2928531