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Copyright by Eunyoung Park 2007 - The University of Texas at Austin

Copyright by Eunyoung Park 2007 - The University of Texas at Austin

FIGURE 3.5. MMP

FIGURE 3.5. MMP ANTIBODIES BLOCK THE INVASION OF HCT-116 AND SW620 CELLS. HCT-116 (A) and SW620 (B) cells were serum starved for 48 h before seeding at a density of 1 x 10 5 cells/well on Matrigel-coated Boyden chambers. The upper portion of the chambers contained 10 μg/ml of MMP-2 or -9 antibody in serum free media. The lower chamber contained 20 ng/ml HGF and 10% FBS. Cell invasion was measured after 24 h by propidium iodide staining as described in Materials and Methods. All data are reported as mean ± SEM, n=2. *Significantly different from control, P < 0.05. 67

FIGURE 3.6. EFFECT OF RETINOL ON TIMP-1 PROTEIN AND MRNA. HCT-116 and SW620 cells were treated with 0 (ethanol vehicle control), 1, or 10 μM retinol for 24 h. Proteins were collected from HCT-116 (A) and SW620 (B) cells and culture media as described in Materials and Methods. TIMP-1 protein was detected in cell lysates and conditioned media. β-Actin is shown as a loading control for the cell lysates. TIMP-1 mRNA levels in HCT-116 (C) and SW620 (D) cells were detected by quantitative real time RT-PCR. Quantitative real time RT-PCR reactions were performed in duplicate. Data are reported as mean ± SEM, n=3. Relative amounts of TIMP-1 cDNA were calculated using the comparative CT method. CT values obtained for TIMP-1 were normalized to corresponding CT values of GAPDH. *Significantly different from control, P < 0.05. 68

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