Mechanical stiffness augments ligand-dependent progesterone receptor B activation via MEK 1/2 and Rho/ROCK-dependent signaling pathways in uterine fibroid cells.

Abstract

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.

• Authors: Christina N Cordeiro Mitchell, Md Soriful Islam, Sadia Afrin, Joshua Brennan, Kevin J Psoter, James H Segars
• Journal: Fertility and sterility (Fertil Steril) Vol. 116 Issue 1 Pg. 255-265 (07 2021) ISSN: 1556-5653 [Electronic] United States
• PMID: 33676751 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2020. Published by Elsevier Inc.

Chemical References

• Ligands
• Polystyrenes
• Protein Kinase Inhibitors
• Receptors, Progesterone
• Silicones
• progesterone receptor B
• Progesterone
• MAP2K2 protein, human
• rho-Associated Kinases
• MAP Kinase Kinase 1
• MAP Kinase Kinase 2
• MAP2K1 protein, human
• rho GTP-Binding Proteins

Topics

• Cell Culture Techniques
• Cell Line, Tumor
• Extracellular Matrix (genetics, metabolism, pathology)
• Female
• Humans
• Leiomyoma (enzymology, genetics, pathology)
• Ligands
• MAP Kinase Kinase 1 (antagonists & inhibitors, metabolism)
• MAP Kinase Kinase 2 (antagonists & inhibitors, metabolism)
• Mechanotransduction, Cellular (drug effects)
• Polystyrenes (chemistry)
• Progesterone (pharmacology)
• Protein Kinase Inhibitors (pharmacology)
• Receptors, Progesterone (agonists, metabolism)
• Silicones (chemistry)
• Uterine Neoplasms (enzymology, genetics, pathology)
• rho GTP-Binding Proteins (antagonists & inhibitors, metabolism)
• rho-Associated Kinases (antagonists & inhibitors, metabolism)