Journal: Biological Chemistry

Article Title: Peritumoral administration of GPI-anchored TIMP-1 inhibits colon carcinoma growth in Rag-2 γ chain-deficient mice

doi: 10.1515/bc.2009.098

Figure Lengend Snippet: Figure 2 The GPI-anchored TIMP-1 suppresses proliferation of SW480 cells and inhibits tumor invasion. The effect of cell surface engineering with TIMP-1-GPI on SW480 invasion through the Matrigel basement membrane model was assessed. Optimal invasion of the SW480 cells in response to 10% FCS, 4 ng/ml EGF, or 4 ng/ml VEGF were set to ‘zero’, and the 100% inhibition value was set to the migration level observed with untreated SW480 cells (Djafarzadeh et al., 2004). 5=103 SW480 cells per 100 ml medium were cultured in 96-well microtiter plates for 24 h under standard conditions to yield firmly attached and stably growing cells (Djafarzadeh et al., 2006). The SW480 cells were then pretreated with 2, 4, 6, 8, 10, 12, and 14 ng/ml of TIMP-1-GPI or 14 ng/ml of rhTIMP-1 as described previously (Djafarzadeh et al., 2006). After 1 h the cells were washed. (A) The effect of increasing levels of TIMP-1-GPI or rhTIMP-1 control protein on the proliferation of SW480 was measured using an MTT assay (Djafarzadeh et al., 2006). MTT was added after 24 h, 48 h, or 72 h as indicated. (B) The effect on the ability of SW480 to invade Matrigel was measured using a modified Boyden chamber as described earlier (Djafarzadeh et al., 2004, 2006).

Article Snippet: The effect of TIMP-1-GPI vs. rhTIMP-1 treatment on the ability of SW480 cells to invade Matrigel was measured using induced migration to fetal calf serum (FCS) (Biocrom, Berlin, Germany), vascular endothelial growth factor (VEGF) (R&D systems, Minneapolis, USA), and EGF (Biochrom, Berlin, Germany) using a modified Boyden chamber (Djafarzadeh et al., 2004, 2006).

Techniques: Membrane, Inhibition, Migration, Cell Culture, Stable Transfection, Control, MTT Assay, Modification