N-cadherin knock-down decreases invasiveness of esophageal squamous cell carcinoma in vitro

Figure 1 Expression of E-cadherin and N-cadherin in ESCC tissues and normal tissues. A: Strong expression (yellowish brown) of E-cadherin on the membrane of normal esophageal epithelial cells (PV, × 200); B: Weak expression (yellow) of E-cadherin on the membrane of ESCC cells (PV, × 200); C: Weak expression (yellow) of N-cadherin in the cytoplasm of normal esophageal epithelial cells (PV, × 200); D: Strong expression (yellowish brown) of N-cadherin in the cytoplasm of ESCC cells (PV, × 200).

Figure 2 EGFP expression in PT67 cells and EC9706 cells after pMSCVneo/N-cadherin transfection. A: Selected by 1000 mg/L G418 for 15 d and 300 mg/L G418 for 15 d, the PT-67 cells transfected with pMSCVneo/N-cadherin plasmid expressed EGFP stably. The figure was taken under fluorescence microscope (× 400, at 488 nm); B: Selected by 600 mg/L G418 for 14 d and 300 mg/L G418 for 10 d, the EC9706 cells infected with pMSCVneo/N-cadherin viral supernatant expressed EGFP stably. The figure was taken under fluorescence microscope (× 400, at 488 nm).

Figure 3 E-cadherin, N-cadherin and MMP-9 gene transcripts detected by real-time PCR. 1: Positive control; 2: Untreated EC9706 cells; 3: EC9706 cells with control vector; 4: EC9706 cells with N-cadherin RNAi. ^aP < 0.05 vs 2 and 3.

Figure 4 E-cadherin, N-cadherin and MMP-9 proteins detected by Western bloting. 1: Positive control; 2: Untreated EC9706 cells; 3: EC9706 cells with control vector; 4: EC9706 cells with N-cadherin RNAi. E-cadherin proteins were 0.247 ± 0.010, 0.252 ± 0.087, 0.249 ± 0.07 and 0.250 ± 0.006, respectively, from lane 1 to lane 4. P > 0.05. The N-cadherin proteins were 0.681 ± 0.003, 0.679 ± 0.004, 0.653 ± 0.009 and 0.342 ± 0.006, respectively, from lane 1 to lane 4. P < 0.05. The MMP-9 proteins were 0.624 ± 0.011, 0.628 ± 0.010, 0.623 ± 0.009 and 0.282 ± 0.010, respectively, from lane 1 to lane 4. P < 0.05.

Figure 5 Migration assay in transwell chamber. A: Untreated EC9706 cells; B: EC9706 cells with control vector; C: EC9706 cells with N-cadherin RNAi. They were 123.40 ± 8.234, 126.00 ± 10.295 and 49.60 ± 6.804, respectively, from A to C. P < 0.05. (HE, × 200).