TY - JOUR
T1 - Mechanical stiffness augments ligand-dependent progesterone receptor B activation via MEK 1/2 and Rho/ROCK–dependent signaling pathways in uterine fibroid cells
AU - Cordeiro Mitchell, Christina N.
AU - Islam, Md Soriful
AU - Afrin, Sadia
AU - Brennan, Joshua
AU - Psoter, Kevin J.
AU - Segars, James H.
N1 - Funding Information:
C.N.C.M was supported in part by the Edward E. Wallach Research Award. M.S.I. was supported in part by the Ines Mandl Research Foundation. S.A. has nothing to disclose. J.B. has nothing to disclose. J.H.S. was supported in part by the Howard and Georgeanna Jones Endowment. K.J.P. has nothing to disclose.
Publisher Copyright:
© 2020
PY - 2021/7
Y1 - 2021/7
N2 - Objective: To test whether mechanical substrate stiffness would influence progesterone receptor B (PRB) signaling in fibroid cells. Uterine fibroids feature an excessive extracellular matrix, increased stiffness, and altered mechanical signaling. Fibroid growth is stimulated by progestins and opposed by anti-progestins, but a functional interaction between progesterone action and mechanical signaling has not been evaluated. Design: Laboratory studies. Setting: Translational science laboratory. Patient(s)/Animal(s): Human fibroid cell lines and patient-matched fibroid and myometrial cell lines. Intervention(s): Progesterone receptor B–dependent reporter assays and messenger RNA quantitation in cells cultured on stiff polystyrene plates (3GPa) or soft silicone plates (930KPa). Pharmacologic inhibitors of extracellular signal-related protein kinase (ERK) kinase 1/2 (MEK 1/2; PD98059), p38 mitogen-activated protein kinase (SB202190), receptor tyrosine kinases (RTKs; nintedanib), RhoA (A13), and Rho-associated coiled-coil kinase (ROCK; Y27632). Main Outcome Measure(s): Progesterone-responsive reporter activation. Result(s): Fibroid cells exhibited higher PRB-dependent reporter activity with progesterone (P4) in cells cultured on stiff vs. soft plates. Mechanically induced PRB activation with P4 was decreased 62% by PD98059, 78% by nintedanib, 38% by A13, and 50% by Y27632. Overexpression of the Rho-guanine nucleotide exchange factor (Rho-GEF), AKAP13, significantly increased PRB-dependent reporter activity. Collagen 1 messenger RNA levels were higher in fibroid cells grown on stiff vs. soft plates with P4. Conclusion(s): Cells cultured on mechanically stiff substrates had enhanced PRB activation via a mechanism that required MEK 1/2 and AKAP13/RhoA/ROCK signaling pathways. These studies provide a framework to explore the mechanisms by which mechanical stiffness affects progesterone receptor activation.
AB - Objective: To test whether mechanical substrate stiffness would influence progesterone receptor B (PRB) signaling in fibroid cells. Uterine fibroids feature an excessive extracellular matrix, increased stiffness, and altered mechanical signaling. Fibroid growth is stimulated by progestins and opposed by anti-progestins, but a functional interaction between progesterone action and mechanical signaling has not been evaluated. Design: Laboratory studies. Setting: Translational science laboratory. Patient(s)/Animal(s): Human fibroid cell lines and patient-matched fibroid and myometrial cell lines. Intervention(s): Progesterone receptor B–dependent reporter assays and messenger RNA quantitation in cells cultured on stiff polystyrene plates (3GPa) or soft silicone plates (930KPa). Pharmacologic inhibitors of extracellular signal-related protein kinase (ERK) kinase 1/2 (MEK 1/2; PD98059), p38 mitogen-activated protein kinase (SB202190), receptor tyrosine kinases (RTKs; nintedanib), RhoA (A13), and Rho-associated coiled-coil kinase (ROCK; Y27632). Main Outcome Measure(s): Progesterone-responsive reporter activation. Result(s): Fibroid cells exhibited higher PRB-dependent reporter activity with progesterone (P4) in cells cultured on stiff vs. soft plates. Mechanically induced PRB activation with P4 was decreased 62% by PD98059, 78% by nintedanib, 38% by A13, and 50% by Y27632. Overexpression of the Rho-guanine nucleotide exchange factor (Rho-GEF), AKAP13, significantly increased PRB-dependent reporter activity. Collagen 1 messenger RNA levels were higher in fibroid cells grown on stiff vs. soft plates with P4. Conclusion(s): Cells cultured on mechanically stiff substrates had enhanced PRB activation via a mechanism that required MEK 1/2 and AKAP13/RhoA/ROCK signaling pathways. These studies provide a framework to explore the mechanisms by which mechanical stiffness affects progesterone receptor activation.
KW - Progesterone signaling
KW - mechanotransduction
KW - nonclassical signaling
KW - progesterone receptor B
KW - uterine leiomyoma
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U2 - 10.1016/j.fertnstert.2020.12.011
DO - 10.1016/j.fertnstert.2020.12.011
M3 - Article
C2 - 33676751
AN - SCOPUS:85102013189
SN - 0015-0282
VL - 116
SP - 255
EP - 265
JO - Fertility and sterility
JF - Fertility and sterility
IS - 1
ER -