Total protein extraction and bicinchoninic acid protein assay kits were purchased from Nanjing KeyGen Biotech Co., Ltd. (Nanjing, China). Polyvinylidene difluoride membranes were purchased from Merck KGaA. Antibodies against PRL-3 (ab50276; 1:400), AKT (ab8805; 1:500), p-AKT S473 (ab81283; 1:1,000), MMP-2 (ab97779; 1:1,000) and MMP-9 (ab137867; 1:1,000) were purchased from Abcam (Cambridge, UK). The antibody for β-actin (60008-1-Ig; 1:5,000) was purchased from Wuhan Sanying Biotechnology (Wuhan, China). These antibodies were used in the present study was described previously (23 (link)–26 (link)). Membranes were incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibody (ab6721; 1:3,000) or goat anti-mouse secondary antibody (Wuhan Sanying Biotechnology; SA00012-6; 1:2,000) and the western blotting protocol were performed as previously described (21 (link)). Reactive protein was detected by an enhanced chemiluminescence kit (EMD Millipore, Billerica, MA, USA). Band intensity was quantified by using the Quantity One image analysis software version 4.62 (Bio-Rad Laboratories, Inc., Hercules, CA, USA).