Versican Proteolysis by ADAMTS Proteases and Its Influence on Sex Steroid Receptor Expression in Uterine Leiomyoma.
Leiomyomas have abundant extracellular matrix (ECM), with upregulation of versican, a large proteoglycan.



We investigated ADAMTS (a disintegrin-like and metalloprotease with thrombospondin type 1 motifs) protease-mediated versican cleavage in myometrium and leiomyoma and the effect of versican knockdown in leiomyoma cells.



We used quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blotting, immunohistochemistry, and RNA in situ hybridization for analysis of myometrium, leiomyoma and immortalized myometrium and leiomyoma cells. Short interfering RNA (siRNA) was used to knockdown versican in leiomyoma cells.



This study was performed in an academic laboratory.



Study subjects were women with symptomatic or asymptomatic leiomyoma.



We quantified messenger RNAs (mRNAs) for versican splice variants. We identified ADAMTS-cleaved versican in myometrium and leiomyoma and ADAMTS messenger RNAs and examined the effect of VCAN siRNA on smooth muscle differentiation and expression of estrogen and progesterone receptors.



The women in the symptomatic group (n = 7) had larger leiomyoma (P = 0.01), heavy menstrual bleeding (P < 0.01), and lower hemoglobin levels (P = 0.02) compared with the asymptomatic group (n = 7), but were similar in age and menopausal status. Versican V0 and V1 isoforms were upregulated in the leiomyomas of symptomatic versus asymptomatic women (P = 0.03 and P = 0.04, respectively). Abundant cleaved versican was detected in leiomyoma and myometrium, as well as in myometrial and leiomyoma cell lines. ADAMTS4 (P = 0.03) and ADAMTS15 (P = 0.04) were upregulated in symptomatic leiomyomas. VCAN siRNA did not effect cell proliferation, apoptosis, or smooth muscle markers, but reduced ESR1 and PR-A expression (P = 0.001 and P = 0.002, respectively).



Versican in myometrium, leiomyomas and in the corresponding immortalized cells is cleaved by ADAMTS proteases. VCAN siRNA suppresses production of estrogen receptor 1 and progesterone receptor-A. These findings have implications for leiomyoma growth.
Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.|Department of Obstetrics and Gynecology and Women's Health Institute, Cleveland Clinic, Cleveland, Ohio 44195.
Ndeye-Aicha
NA
Gueye
Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
Timothy J
TJ
Mead
Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.|Department of Chemistry, Cleveland State University, Cleveland, Ohio 44115.
Christopher D
CD
Koch
Department of Anatomic Pathology, Cleveland Clinic, Cleveland, Ohio 44195.
Charles V
CV
Biscotti
Department of Obstetrics and Gynecology and Women's Health Institute, Cleveland Clinic, Cleveland, Ohio 44195.
Tommaso
T
Falcone
Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio 44195.
Suneel S
SS
Apte
D000071096:ADAMTS Proteins / Q000235:genetics* / Q000378:metabolism; D000071121:ADAMTS4 Protein / Q000235:genetics* / Q000378:metabolism; D000328:Adult; D017209:Apoptosis / Q000235:genetics; D058070:Asymptomatic Diseases; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics; D047628:Estrogen Receptor alpha / Q000235:genetics* / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D055785:Gene Knockdown Techniques; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007889:Leiomyoma / Q000378:metabolism* / Q000473:pathology; D008595:Menorrhagia / Q000209:etiology; D008875:Middle Aged; D009215:Myometrium / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics; D059748:Proteolysis; D012333:RNA, Messenger / Q000378:metabolism*; D034741:RNA, Small Interfering; D011980:Receptors, Progesterone / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D047368:Tumor Burden; D015854:Up-Regulation; D014594:Uterine Neoplasms / Q000378:metabolism* / Q000473:pathology; D053675:Versicans / Q000235:genetics / Q000378:metabolism*
C506487:ESR1 protein, human; D047628:Estrogen Receptor alpha; D006454:Hemoglobins; D020033:Protein Isoforms; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011980:Receptors, Progesterone; C508322:VCAN protein, human; C101118:progesterone receptor A; D053675:Versicans; D000071096:ADAMTS Proteins; C500267:ADAMTS15 protein, human; D000071121:ADAMTS4 Protein; C000603918:ADAMTS4 protein, human