Total protein from tissue specimens and cells was extracted by sample buffer (62.5 mmol/L Tris-HCl, pH 6.8, 2% SDS, 10% glycerol, and 5% 2-β-mercaptoethanol, Sigma-Aldrich, USA). The concentrations of protein were then determined using the BCA protein assay kit (Beyotime, China). Western blot analysis was performed as described 14 (link). Briefly, after being separated in SDS-PAGE gels, proteins were transferred to nitrocellulose membranes (Bio-Rad, USA). The membranes were then incubated with different primary antibodies at 4°C, overnight. Afterwards, respective secondary antibodies were applied at room temperature for 1hour. ECL development solution was manipulated for the visualization of the expression of different protein. The antibodies used are listed as follow: anti-HIF-1α (dilution at 1:1000, A11945, Abclonal, China), anti-FAP (dilution at 1:800, BM5121, Boster, China), anti-E-cadherin (dilution at 1:1000, 3195, CST, USA), anti-Twist (dilution at 1:1000, ab175430, Abcam, England), anti-Snail (dilution at 1:1000, ab53519, Abcam, England).