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 (α-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.

Cancer-associated fibroblasts of the prostate promote a compliant and more invasive phenotype in benign prostate epithelial cells

“…It is expected though that the mechanical properties of the cell cortex in a spread and a suspended state vary, as the cortical thickness depends on cell spreading and is altered on de-adhesion of cells [ 72 ]. Recently, pancreatic CAFs were reported to exhibit a lowered elastic modulus compared with native fibroblasts from the pancreas, although similar changes in the stress fiber alignment as in our study were observed [ 73 , 74 ]. There are, however, some differences in the experimental setup.…”

“…In contrast, Sylianou et al. [ 73 , 74 ] probed non-confluent pancreatic fibroblasts that were immortalized. While organ-specific differences might not be excluded, we also detected patient-to-patient variability in our experiments.…”

Comparison of cytotoxicity of black phosphorus nanosheets in different types of fibroblasts

“…We have selected fibroblast cells for the in vitro cytotoxicity studies because they are heterogeneous and dynamic cell lineage presenting with different populations between human tissues [18, 19]. Fibroblast cells can produce extracellular matrix and collagen, which play critical roles in wound healing [20]. Cancerous cells have been found more sensitive towards BPNSs exposure than normal cells, which is evident from cell viability assays.…”

Type II Collagen from Cartilage of Acipenser baerii Promotes Wound Healing in Human Dermal Fibroblasts and in Mouse Skin

“…Xie et al demonstrated a lower proliferative rate of human mesenchymal stem cells on collagen with stiff fibers than that with soft fibers [ 33 ]. In addition, Stylianou et al reported that an increase of collagen concentration elevated fiber density, stiffness, and contributed to an increase in cell spreading of fibroblasts [ 34 ]. In our study, we found the proliferation of HDFa was decreased by coating SCC at high concentrations, whereas the migration and invasion ability was increased, maybe due to the substrate stiffening.…”