MilliporeSigma
  • Subendothelial stiffness alters endothelial cell traction force generation while exerting a minimal effect on the transcriptome.

Subendothelial stiffness alters endothelial cell traction force generation while exerting a minimal effect on the transcriptome.

Scientific reports (2019-12-05)
Effie E Bastounis, Yi-Ting Yeh, Julie A Theriot
ABSTRACT

Endothelial cells respond to changes in subendothelial stiffness by altering their migration and mechanics, but whether those responses are due to transcriptional reprogramming remains largely unknown. We measured traction force generation and also performed gene expression profiling for two endothelial cell types grown in monolayers on soft or stiff matrices: primary human umbilical vein endothelial cells (HUVEC) and immortalized human microvascular endothelial cells (HMEC-1). Both cell types respond to changes in subendothelial stiffness by increasing the traction stresses they exert on stiffer as compared to softer matrices, and exhibit a range of altered protein phosphorylation or protein conformational changes previously implicated in mechanotransduction. However, the transcriptome has only a minimal role in this conserved biomechanical response. Only few genes were differentially expressed in each cell type in a stiffness-dependent manner, and none were shared between them. In contrast, thousands of genes were differentially regulated in HUVEC as compared to HMEC-1. HUVEC (but not HMEC-1) upregulate expression of TGF-β2 on stiffer matrices, and also respond to application of exogenous TGF-β2 by enhancing their endogenous TGF-β2 expression and their cell-matrix traction stresses. Altogether, these findings provide insights into the relationship between subendothelial stiffness, endothelial mechanics and variation of the endothelial cell transcriptome, and reveal that subendothelial stiffness, while critically altering endothelial cells' mechanical behavior, minimally affects their transcriptome.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Transforming Growth Factor-β2 human, TGF-β2, recombinant, expressed in NSO cells, powder, suitable for cell culture
Sigma-Aldrich
Anti-Disialoganglioside GD3 Antibody, clone MB3.6, clone MB3.6, Chemicon®, from mouse
Sigma-Aldrich
hEGF, EGF, recombinant, expressed in E. coli, lyophilized powder, suitable for cell culture
Sigma-Aldrich
Hydrocortisone, BioReagent, suitable for cell culture
Sigma-Aldrich
Acrylamide solution, 40%, suitable for electrophoresis, sterile-filtered
Sigma-Aldrich
Collagen, Type I solution from rat tail, BioReagent, suitable for cell culture, sterile-filtered