Collagen content and extracellular matrix cause cytoskeletal remodelling in pancreatic fibroblasts

Andreas Stylianou, Vasiliki Gkretsi, Maria Louca, Lefteris C. Zacharia, Triantafyllos Stylianopoulos

Research output: Contribution to journalArticle

Abstract

In many solid tumours a desmoplastic reaction takes place, which results in tumour tissue stiffening due to the extensive production of extracellular matrix (ECM) proteins, such as collagen, by stromal cells, mainly fibroblasts (FBs) and cancer-associated fibroblasts (CAFs). In this study, we investigated the effect of collagen stiffness on pancreatic FBs and CAFs, particularly on specific cytoskeleton properties and gene expression involved in tumour invasion. We found that cells become stiffer when they are cultured on stiff substrates and express higher levels of alpha-smooth muscle actin (a-SMA). Also, it was confirmed that on stiff substrates, CAFs are softer than FBs, while on soft substrates they have comparable Young's moduli. Furthermore, the number of spread FBs and CAFs was higher in stiffer substrates, which was also confirmed by Ras-related C3 botulinum toxin substrate 1 (RAC1) mRNA expression, which mediates cell spreading. Although stress fibres in FBs become more oriented on stiff substrates, CAFs have oriented stress fibres regardless of substrate stiffness. Subsequently, we demonstrated that cells' invasion has a differential response to stiffness, which was associated with regulation of Ras homologue family member (RhoA) and Rho-associated, coiled-coil containing protein kinase 1 (ROCK-1) mRNA expression. Overall, our results demonstrate that collagen stiffness modulates FBs and CAFs cytoskeleton remodelling and alters their invasion properties.

Original languageEnglish
Article number0226
JournalJournal of the Royal Society Interface
Volume16
Issue number154
DOIs
Publication statusPublished - 1 Jan 2019

Fingerprint

Fibroblasts
Collagen
Extracellular Matrix
Stress Fibers
Cytoskeleton
Substrates
rac1 GTP-Binding Protein
Stiffness
Tumors
rho-Associated Kinases
Neoplasms
Messenger RNA
Elastic Modulus
Extracellular Matrix Proteins
Stromal Cells
Pancreatic Neoplasms
Smooth Muscle
Cancer-Associated Fibroblasts
Actins
Proteins

Keywords

  • Atomic force microscopy
  • Biomechanics
  • Cancer-associated fibroblasts
  • Collagen
  • Tumour desmoplasia

Cite this

Stylianou, Andreas ; Gkretsi, Vasiliki ; Louca, Maria ; Zacharia, Lefteris C. ; Stylianopoulos, Triantafyllos. / Collagen content and extracellular matrix cause cytoskeletal remodelling in pancreatic fibroblasts. In: Journal of the Royal Society Interface. 2019 ; Vol. 16, No. 154.
@article{d2c613b004374dfa8509c7c6b1d9d2b4,
title = "Collagen content and extracellular matrix cause cytoskeletal remodelling in pancreatic fibroblasts",
abstract = "In many solid tumours a desmoplastic reaction takes place, which results in tumour tissue stiffening due to the extensive production of extracellular matrix (ECM) proteins, such as collagen, by stromal cells, mainly fibroblasts (FBs) and cancer-associated fibroblasts (CAFs). In this study, we investigated the effect of collagen stiffness on pancreatic FBs and CAFs, particularly on specific cytoskeleton properties and gene expression involved in tumour invasion. We found that cells become stiffer when they are cultured on stiff substrates and express higher levels of alpha-smooth muscle actin (a-SMA). Also, it was confirmed that on stiff substrates, CAFs are softer than FBs, while on soft substrates they have comparable Young's moduli. Furthermore, the number of spread FBs and CAFs was higher in stiffer substrates, which was also confirmed by Ras-related C3 botulinum toxin substrate 1 (RAC1) mRNA expression, which mediates cell spreading. Although stress fibres in FBs become more oriented on stiff substrates, CAFs have oriented stress fibres regardless of substrate stiffness. Subsequently, we demonstrated that cells' invasion has a differential response to stiffness, which was associated with regulation of Ras homologue family member (RhoA) and Rho-associated, coiled-coil containing protein kinase 1 (ROCK-1) mRNA expression. Overall, our results demonstrate that collagen stiffness modulates FBs and CAFs cytoskeleton remodelling and alters their invasion properties.",
keywords = "Atomic force microscopy, Biomechanics, Cancer-associated fibroblasts, Collagen, Tumour desmoplasia",
author = "Andreas Stylianou and Vasiliki Gkretsi and Maria Louca and Zacharia, {Lefteris C.} and Triantafyllos Stylianopoulos",
year = "2019",
month = "1",
day = "1",
doi = "10.1098/rsif.2019.0226",
language = "English",
volume = "16",
journal = "Journal of the Royal Society Interface",
issn = "1742-5689",
publisher = "Royal Society of London",
number = "154",

}

Collagen content and extracellular matrix cause cytoskeletal remodelling in pancreatic fibroblasts. / Stylianou, Andreas; Gkretsi, Vasiliki; Louca, Maria; Zacharia, Lefteris C.; Stylianopoulos, Triantafyllos.

In: Journal of the Royal Society Interface, Vol. 16, No. 154, 0226, 01.01.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Collagen content and extracellular matrix cause cytoskeletal remodelling in pancreatic fibroblasts

AU - Stylianou, Andreas

AU - Gkretsi, Vasiliki

AU - Louca, Maria

AU - Zacharia, Lefteris C.

AU - Stylianopoulos, Triantafyllos

PY - 2019/1/1

Y1 - 2019/1/1

N2 - In many solid tumours a desmoplastic reaction takes place, which results in tumour tissue stiffening due to the extensive production of extracellular matrix (ECM) proteins, such as collagen, by stromal cells, mainly fibroblasts (FBs) and cancer-associated fibroblasts (CAFs). In this study, we investigated the effect of collagen stiffness on pancreatic FBs and CAFs, particularly on specific cytoskeleton properties and gene expression involved in tumour invasion. We found that cells become stiffer when they are cultured on stiff substrates and express higher levels of alpha-smooth muscle actin (a-SMA). Also, it was confirmed that on stiff substrates, CAFs are softer than FBs, while on soft substrates they have comparable Young's moduli. Furthermore, the number of spread FBs and CAFs was higher in stiffer substrates, which was also confirmed by Ras-related C3 botulinum toxin substrate 1 (RAC1) mRNA expression, which mediates cell spreading. Although stress fibres in FBs become more oriented on stiff substrates, CAFs have oriented stress fibres regardless of substrate stiffness. Subsequently, we demonstrated that cells' invasion has a differential response to stiffness, which was associated with regulation of Ras homologue family member (RhoA) and Rho-associated, coiled-coil containing protein kinase 1 (ROCK-1) mRNA expression. Overall, our results demonstrate that collagen stiffness modulates FBs and CAFs cytoskeleton remodelling and alters their invasion properties.

AB - In many solid tumours a desmoplastic reaction takes place, which results in tumour tissue stiffening due to the extensive production of extracellular matrix (ECM) proteins, such as collagen, by stromal cells, mainly fibroblasts (FBs) and cancer-associated fibroblasts (CAFs). In this study, we investigated the effect of collagen stiffness on pancreatic FBs and CAFs, particularly on specific cytoskeleton properties and gene expression involved in tumour invasion. We found that cells become stiffer when they are cultured on stiff substrates and express higher levels of alpha-smooth muscle actin (a-SMA). Also, it was confirmed that on stiff substrates, CAFs are softer than FBs, while on soft substrates they have comparable Young's moduli. Furthermore, the number of spread FBs and CAFs was higher in stiffer substrates, which was also confirmed by Ras-related C3 botulinum toxin substrate 1 (RAC1) mRNA expression, which mediates cell spreading. Although stress fibres in FBs become more oriented on stiff substrates, CAFs have oriented stress fibres regardless of substrate stiffness. Subsequently, we demonstrated that cells' invasion has a differential response to stiffness, which was associated with regulation of Ras homologue family member (RhoA) and Rho-associated, coiled-coil containing protein kinase 1 (ROCK-1) mRNA expression. Overall, our results demonstrate that collagen stiffness modulates FBs and CAFs cytoskeleton remodelling and alters their invasion properties.

KW - Atomic force microscopy

KW - Biomechanics

KW - Cancer-associated fibroblasts

KW - Collagen

KW - Tumour desmoplasia

UR - http://www.scopus.com/inward/record.url?scp=85066491632&partnerID=8YFLogxK

U2 - 10.1098/rsif.2019.0226

DO - 10.1098/rsif.2019.0226

M3 - Article

VL - 16

JO - Journal of the Royal Society Interface

JF - Journal of the Royal Society Interface

SN - 1742-5689

IS - 154

M1 - 0226

ER -