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Transforming Growth Factor β3 Regulates the Versican Variants in the Extracellular Matrix-Rich Uterine Leiomyomas

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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 β (TGF-β) signaling. Here we characterized TGF-β3 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-β3. 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 β3 transcripts were increased 4-fold in leiomyoma versus matched myometrium. Phosphorylated-TGF-β 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-β3 increased GAG-rich versican variant expression 7 to 12 fold. Neutralizing TGF-β3 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-β signaling pathway and excessive production of GAG-rich versican variants.

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Authors and Affiliations

  1. Department of Obstetrics & Gynecology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, A-3078, 20814, Bethesda, MD, USA

    John M. Norian MD, Minnie Malik PhD, Candace Y. Parker MD, Doina Joseph MS, James H. Segars MD & William H. Catherino MD, PhD

  2. Reproductive Biology and Medicine Branch, The Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA

    John M. Norian MD, Candace Y. Parker MD, James H. Segars MD & William H. Catherino MD, PhD

  3. Department of Obstetrics and Gynecology, Duke University School of Medicine, Durham, North Carolina, USA

    Phyllis C. Leppert MD, PhD

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  1. John M. Norian MD
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  2. Minnie Malik PhD
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Correspondence to William H. Catherino MD, PhD.

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The work was performed at the Department of Obstetrics and Gynecology, Uniformed Services University of the Health Sciences, Bethesda, Maryland.

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Norian, J.M., Malik, M., Parker, C.Y. et al. Transforming Growth Factor β3 Regulates the Versican Variants in the Extracellular Matrix-Rich Uterine Leiomyomas. Reprod. Sci. 16, 1153–1164 (2009). https://doi.org/10.1177/1933719109343310

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