TY - JOUR
T1 - 2-Methoxyestradiol causes functional repression of transforming growth factor β3 signaling by ameliorating Smad and non-Smad signaling pathways in immortalized uterine fibroid cells
AU - Salama, Salama A.
AU - Diaz-Arrastia, Concepcion R.
AU - Kilic, Gokhan S.
AU - Kamel, Marwa W.
N1 - Funding Information:
Supported in part by an Alliance for Nanohealth grant to Salama A. Salama, start-up funds from Baylor College of Medicine to Concepcion R. Diaz-Arrastia, and a Chairman's Departmental Faculty Development grant to Gokhan S. Kilic.
PY - 2012/7
Y1 - 2012/7
N2 - Objective: To investigate the effects and the mechanism of action of 2-methoxyestradiol (2ME2) on transforming growth factor (TGF) β3-induced profibrotic response in immortalized human uterine fibroid smooth muscle (huLM) cells. Design: Laboratory study. Setting: University research laboratory. Patients(s): Not applicable. Interventions(s): Not applicable. Main Outcome Measure(s): huLM cells were treated with TGF-β3 (5 ηg/mL) in the presence or absence of specific Smad3 inhibitor SIS3 (1 μmol/L), inhibitor of the PI3K/Akt (LY294002, 10 μmol/L), or 2ME 2 (0.5 μmol/L), and the expression of collagen (Col) type I(αI), Col III(αI), plasminogen activator inhibitor (PAI) 1, connective tissue growth factor (CTGF), and α-smooth muscle actin (α-SMA) were determined by real-time reverse-transcription polymerase chain reaction and immunoblotting. The effect of 2ME2 on Smad-microtubule binding was evaluated by coimmunoprecipitation. Result(s): Our data revealed that TGF-β3-induced fibrogenic response in huLM is mediated by both Smad-dependent and Smad-independent PI3K/Akt/mTOR signaling pathways. 2ME2 abrogates TGF-β3-induced expression of Col I(αI), Col III(αI), PAI-1, CTGF, and α-SMA. Molecularly, 2ME2 ameliorates TGF-β3-induced Smad2/3 phosphorylation and nuclear translocation. In addition, 2ME2 inhibits TGF-β3-induced activation of the PI3K/Akt/mTOR pathway. Conclusion(s): TGF-β3-induced profibrotic response in fibroid cells is mediated by Smad-dependent and Smad-independent PI3K/Akt/mTOR pathways. 2ME2 inhibits TGF-β3 profibrotic effects in huLM cells by ameliorating both Smad-dependent and Smad-independent signaling pathways.
AB - Objective: To investigate the effects and the mechanism of action of 2-methoxyestradiol (2ME2) on transforming growth factor (TGF) β3-induced profibrotic response in immortalized human uterine fibroid smooth muscle (huLM) cells. Design: Laboratory study. Setting: University research laboratory. Patients(s): Not applicable. Interventions(s): Not applicable. Main Outcome Measure(s): huLM cells were treated with TGF-β3 (5 ηg/mL) in the presence or absence of specific Smad3 inhibitor SIS3 (1 μmol/L), inhibitor of the PI3K/Akt (LY294002, 10 μmol/L), or 2ME 2 (0.5 μmol/L), and the expression of collagen (Col) type I(αI), Col III(αI), plasminogen activator inhibitor (PAI) 1, connective tissue growth factor (CTGF), and α-smooth muscle actin (α-SMA) were determined by real-time reverse-transcription polymerase chain reaction and immunoblotting. The effect of 2ME2 on Smad-microtubule binding was evaluated by coimmunoprecipitation. Result(s): Our data revealed that TGF-β3-induced fibrogenic response in huLM is mediated by both Smad-dependent and Smad-independent PI3K/Akt/mTOR signaling pathways. 2ME2 abrogates TGF-β3-induced expression of Col I(αI), Col III(αI), PAI-1, CTGF, and α-SMA. Molecularly, 2ME2 ameliorates TGF-β3-induced Smad2/3 phosphorylation and nuclear translocation. In addition, 2ME2 inhibits TGF-β3-induced activation of the PI3K/Akt/mTOR pathway. Conclusion(s): TGF-β3-induced profibrotic response in fibroid cells is mediated by Smad-dependent and Smad-independent PI3K/Akt/mTOR pathways. 2ME2 inhibits TGF-β3 profibrotic effects in huLM cells by ameliorating both Smad-dependent and Smad-independent signaling pathways.
KW - 2-methoxyestradiol
KW - Transforming growth factor β signaling
KW - uterine fibroids
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U2 - 10.1016/j.fertnstert.2012.04.002
DO - 10.1016/j.fertnstert.2012.04.002
M3 - Article
C2 - 22579131
AN - SCOPUS:84863004802
SN - 0015-0282
VL - 98
SP - 178-184.e1
JO - Fertility and Sterility
JF - Fertility and Sterility
IS - 1
ER -