Investigation of tumor-peritoneal interactions in the pathogenesis of peritoneal metastases using a novel ex vivo peritoneal model.
BACKGROUND
Peritoneal metastasis occurs in up to 30% of patients with gastric cancer. The aim of this experimental study is to develop and validate a novel ex vivo model of the human peritoneum to better identify factors involved in the development of peritoneal metastasis in order to improve its management and prognosis.


METHODS
Peritoneal discs harvested from hernia sacs obtained at inguinal hernia surgery were suspended in media using Teflon rings. Viability of the tissue was investigated using MTS assay, light and scanning electron microscopy (LM and SEM) over 72 h. To assess validity of the model, phenotypic changes in tumor cells were investigated. Changes in matrix metalloproteinases (MMP)-2 and -9 activities in HGC and AGS gastric adenocarcinoma cells after co-culture were investigated using zymography. Modulation of tumor cell adhesion to peritoneum after exposure to heparin was assessed using a fluorometric adhesion assay. Analysis was performed using Kruskal-Wallis for multiple comparisons and Mann-Witney U for comparisons between each group.


RESULTS
MTS assay showed reduced viability after 72 h (P = 0.047, compared with 24 h). Mesothelial cell loss at 48 h was demonstrated by LM and SEM, confirming peritoneal viability for at least 24 h after tissue harvesting. Zymography confirmed increased MMP2 and -9 activities in tumor cells and peritoneal tissue during co-culture compared with controls, and heparin significantly reduced tumor cell adherence (P = 0.04), as observed in published in vivo models.


CONCLUSIONS
A validated complete model of peritoneum was developed that has shown potential to determine realistic mechanisms of peritoneal metastasis.
Department of Surgical Oncology and Technology, Imperial College London, St Mary's Hospital, London, United Kingdom. emily.cabourne@googlemail.com
Emily J
EJ
Cabourne
Gretta
G
Roberts
Robert
R
Goldin
Timothy
T
Ryder
Margaret
M
Mobberly
Paul
P
Ziprin
D000230:Adenocarcinoma / Q000201:enzymology / Q000453:epidemiology / Q000401:mortality / Q000473:pathology / Q000601:surgery; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods; D045744:Cell Line, Tumor; D002470:Cell Survival; D018920:Coculture Techniques; D006552:Hernia, Inguinal / Q000473:pathology / Q000601:surgery*; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D008855:Microscopy, Electron, Scanning / Q000379:methods; D009362:Neoplasm Metastasis / Q000473:pathology*; D010534:Peritoneal Neoplasms / Q000473:pathology* / Q000601:surgery*; D010537:Peritoneum / Q000473:pathology / Q000601:surgery / Q000648:ultrastructure; D011379:Prognosis; D013274:Stomach Neoplasms / Q000201:enzymology / Q000453:epidemiology / Q000401:mortality / Q000473:pathology / Q000601:surgery
D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9