Nicotine enhances migration and invasion of human esophageal squamous carcinoma cells which is inhibited by nimesulide

Figure 1 Effect of nicotine on the motility (A) and invasiveness (B) of the esophageal squamous carcinoma cell line TE-13. Numbers of TE-13 cells which migrated through the polycarbonate (A) or matrigel (B) membrane of Transwell^® to the lower surface of the membrane were counted under a light microscope (× 400). Columns, mean of three triplicate experiments; bars, standard error of mean. ^bP < 0.01 vs the control group; ^dP < 0.01 vs the nicotine 100 mg/mL group.

Figure 2 Western blotting analysis. A: Effect of nicotine on COX-2 protein expression in TE-13 cells, 1: DMSO; 2: Nicotine 100 &mgr;g/mL; 3: Nicotine 200 &mgr;g/mL; B: Effect of nimesulide on COX-2 protein expression in TE-13 cells, 1: DMSO; 2: Nicotine 200 &mgr;g/mL; 3: Nicotine 200 &mgr;g/mL + Nimesulide 100 &mgr;mol/mL.

Figure 3 Effect of nimesulide on the motility (A) and invasiveness (B) of the esophageal squamous carcinoma cell line TE-13 induced by nicotine treatment. Numbers of TE-13 cells which migrated through the polycarbonate (A) or matrigel (B) membrane of Transwell^® to the lower surface of the membrane were counted under a light microscope (× 400). Data are expressed as mean ± SE. ^bP < 0.01 vs the control group; ^dP < 0.01 vs the nicotine 200 &mgr;g/mL group.

Figure 4 A Gelatin zymography of MMP-2 activity of TE-13. A: Effect of nicotine on MMP-2 activity of TE-13 cells. Our study showed nicotine increased the activity of MMP-2. 1: DMSO; 2: Nicotine 100 &mgr;g/mL; 3: Nicotine 200 &mgr;g/mL; B: Effect of nimesulide on MMP-2 activity of TE-13 cells treated by nicotine. Our study showed nimesulide suppressed the effect of nicotine that increased the activity of MMP-2. 1: DMSO; 2: Nicotine 200 &mgr;g/mL; 3: Nicotine 200 &mgr;g/mL + Nimesulide 100 mmol/L.

Figure 5 The protein levels of MMP-2 of TE-13 by treatment of nicotine or nicotine and nimesulide were analyzed by ELISA. The experiments were done as described in Materials and Methods. Data are expressed as mean ± SE. ^aP < 0.05 versus the nicotine 200 &mgr;g/mL group.