Abstract
The role of the mesothelial layer in the peritoneal spreading of cancer cells is only partially clarified. Here we attempted to better define the mesothelial contribution to the tumor cell adhesion using a direct adhesion test applied to human primary cultures of mesothelial cells (HPMCs) derived from the peritoneal washes of patients with gastric and colorectal cancers. Gastric and colon carcinoma cells were seeded on different mesothelial monolayers and quantitative fluorescence analysis was performed to analyze their growth and adhesive properties. The adhesion of the cancer cells was not affected by the origin of the HPMCs when derived from patients with different cancers or with benign disease. In contrast, the high levels of ICAM1 expression and ROS production, which characterize these senescent mesothelial cells, enhanced the tumor cell adhesion. These results suggest that the mesothelial adhesive properties are dependent on the cell senescence, while are not affected by the tumor environment. The use of peritoneal washes as a source to isolate HPMCs provides a practical and reliable tool for the in vitro analysis of the mesothelial conditions affecting the peritoneal carcinomatosis.
Highlights
The peritoneal spreading of gastric and colorectal cancers represents a frequent event occurring after curative resection [1,2,3]
Primary cultures and co-cultures Primary cultures of Human Peritoneal Mesothelial Cells (HPMCs) were obtained from intraoperatively peritoneal lavages [18] of patients affected by peritoneal carcinomatosis from colorectal cancer (#062) or gastric cancer (#219) as well as of patients affected by non-cancerous disease (#002), who underwent surgery at the A Unit of Surgery of Sant’Andrea Hospital
After 24 hours from seeding, to recognize the mesothelial cells making up the Met-5A monolayer, we stained the co-cultures with a primary antibody directed against vimentin, a component of the intermediate filaments of the cytoskeleton, followed by a secondary Ab labeled with the FITC fluorocrome: the signal was compatible with the structure and localization of vimentin, which appears as perinuclear cytoplasmic bundles of filaments (Fig. 1B)
Summary
The peritoneal spreading of gastric and colorectal cancers represents a frequent event occurring after curative resection [1,2,3]. For the detailed analysis of the molecular mechanisms affecting the adhesive stage, different in vitro or ex-vivo models have been developed [11,12,13] and primary cultures of mesothelial cells have been obtained to test the adhesion of cancer cells in presence of promoting or interfering agents [8,12]. Most of these models utilize either established cell lines or human primary cultures of mesothelial cells isolated from omental fragments [10,14,15]. It has been proposed that the peritoneal lavages, being the gold standard for assessing the presence of peritoneal dissemination of gastric and colorectal cancer [16,17,18], are a good and more practical source of mesothelial cells to be propagated in vitro [19], their use in co-culture models has not been explored
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