Myometrial cells undergo fibrotic transformation under the influence of transforming growth factor β-3 Doina S. Joseph, M.S., Minnie Malik, Ph.D., Sahadat.

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Myometrial cells undergo fibrotic transformation under the influence of transforming growth factor β-3 Doina S. Joseph, M.S., Minnie Malik, Ph.D., Sahadat Nurudeen, M.D., William H. Catherino, M.D., Ph.D. Fertility and Sterility Volume 93, Issue 5, Pages 1500-1508 (March 2010) DOI: 10.1016/j.fertnstert.2009.01.081 Copyright © 2010 Terms and Conditions

Figure 1 Expression of extracellular matrix genes in myometrial and leiomyoma cells exposed to different concentrations of transforming growth factor (TGF) β3 for 24 hours, as analyzed by quantitative polymerase chain reaction. (A) Collagen 1A1 (COL1A1) was elevated in untreated leiomyoma cells compared with patient-matched myometrial cells. With increasing concentrations of TGF-β3, myometrial COL 1A1 expression increased to levels similar to those in leiomyoma cells. (B) Connective tissue growth factor (CTGF) was slightly elevated in leiomyoma cells, and TGF-β3 treatment resulted in elevated expression in both cell types. (C) Fibronectin was elevated in untreated leiomyoma cells, and TGF-β3 treatment again induced expression in myometrial cells to levels similar to those in leiomyoma cells. Fold expression = gene expression in cell and treatment column divided by gene expression in untreated myometrial cells. ∗P<.05 compared with untreated myometrial cells. ∗∗P<.05 compared with similarly treated leiomyoma cells. Fertility and Sterility 2010 93, 1500-1508DOI: (10.1016/j.fertnstert.2009.01.081) Copyright © 2010 Terms and Conditions

Figure 2 Expression of extracellular matrix proteins in leiomyoma cells exposed to different concentrations of TGF-β3 for 24 hours, as analyzed by Western blot analysis. (A) COL1A1 protein expression increased with TGF-β3 treatment in both myometrial and leiomyoma cells. At TGF-β3 concentrations of ≥1 ng/mL, myometrial COL1A1 expression was similar to that in leiomyoma cells. Similar expression patterns with treatment were found with CTGF (B) and fibronectin (C). Abbreviations as in Figure 1. ∗P<.05 compared with untreated myometrial cells. ∗∗P<.05 compared with similarly treated leiomyoma cells. Fertility and Sterility 2010 93, 1500-1508DOI: (10.1016/j.fertnstert.2009.01.081) Copyright © 2010 Terms and Conditions

Figure 3 Expression of extracellular matrix resorption genes in myometrial and leiomyoma cells exposed to different concentrations of TGF-β3 for 24 hours, as analyzed by quantitative polymerase chain reaction. (A) Matrix metalloproteinase 2 was elevated in untreated leiomyoma cells compared with patient-matched myometrial cells. With increasing concentrations of TGF-β3, myometrial MMP-2 expression decreased in a dose-dependent manner. (B) Matrix metalloproteinase 11 was similarly elevated in leiomyoma cells, and TGF-β3 treatment resulted in decreased expression in both cell types in a dose-dependent manner. Abbreviations as in Figure 1. ∗P<.05 compared with untreated myometrial cells. ∗∗P<.05 compared with similarly treated leiomyoma cells. Fertility and Sterility 2010 93, 1500-1508DOI: (10.1016/j.fertnstert.2009.01.081) Copyright © 2010 Terms and Conditions