Increased migration of colorectal cancer cells induced by TNF-alpha-treated stromal fibroblasts from human liver metastases

Methods Primary cultures of cancer-associated stromal fibroblasts (CAFs) were generated from human tissues harvested during hepatic resection. TNF-alpha expression in tissue was examined by immunohistochemistry. Activation of nuclear-factor-kappaB (NF-kappaB) activity was measured by gel mobility shift assay. The effect of TNF-alpha on migratory capacity and gene expression of CAFs was tested in presence/absence of parthenolide, an herbal inhibitor of NF-kappaB. Gene expression in tissues and cell cultures was examined by Northern blot analysis. Protein measurements in the cell culture supernatant were performed with cytometric capture beads.