In-vitro investigation on the role of pancreatic stellate cells and tumor cells in angiogenesis
Background: Angiogenesis represents a multistep process involving proliferation, migration and tube formation of endothelial cells. In most tumors including pancreatic carcinoma, angiogenesis plays a pivotal role in the cancer progression. Pancreatic stellate cells (PSCs) are the main source of extracellular matrix providing a microenvironment which is advantageous for pancreatic cancer growth and survival. Until now the role of PSCs in angiogenesis is unclear. This study investigated the role of PSCs, pancreatic carcinoma cells (PCCs) and the interaction of both cell types on angiogenesis using cultured human PSCs, PCCs and human umbilical vein endothelial cells (HUVECs). Methods: Supernatants (SNs) of human PSCs and PCCs, cultivated alone or in coculture, were added to cultured HUVECs. Proliferation and viability of HUVEC were measured by EdU incorporation assay and PI/Hoechst staining. Migration of HUVEC was quantified by single cell tracking assay. Tube formation was shown by microscopy after seeding HUVEC on extracellular matrix gel. The metalloproteinase (MMP) inhibitor GM6001 was used to determine the role of MMPs on HUVEC migration. Expression of MMP-2 in PSCs and PCCs was shown by zymography. Results: PSC- and PCC-SNs stimulated proliferation, viability and migration of HUVEC. The effect on migration was dose dependent. SNs of PSC/PCC coculture stimulated HUVEC motility to a higher extent compared to the mixture of SNs of monoculture cells. PSC stimulated tube formation of HUVEC, and the MMP inhibitor GM6001 reduced the migration of HUVEC stimulated by PSC or cocultures of PSC/PCC. Finally, cocultures of PSC/PCC expressed more MMP-2 than PSC and PCC alone. Conclusion: Both PSC and PCC stimulate angiogenic properties of HUVEC in vitro. Via interaction of PSC and PCC angiogenesis stimulating activity is increased. MMP-2 released through interaction of PSC and PCC seems to represent a prominent factor stimulating endothelial cell motility.
Subject HeadingsAngiogenese [GND]
Pancreatic neoplasms [MeSH]
Pancreatic stellate cells [MeSH]