Overexpression of Tripartite Motif Conaining 55 (TRIM55) Inhibits Migration and Invasion of Hepatocellular Carcinoma (HCC) Cells via Epithelial-Mesenchymal Transition and Matrix Metalloproteinase-2 (MMP2)

Li, Xinyu; Huang, Lei; Gao, Weijie

Total proteins in HCC cells were extracted by lysis buffer, and the protein concentration was measured by Bicinchoninic acid (BCA) assay. The SDS-PAGE was conducted according to the manufacturer's protocol. After transfer to a PVDF membrane, membranes were incubated with primary antibodies at 4°C overnight in a buffer containing 5% skim milk. The primary antibodies were as follows: TRIM55 (Novus, NBP2-33691, dilution 1/1000), E-cadherin and Vimentin (Cell Signaling Technology, (EMT) Antibody Sampler Kit #9782, dilution 1/1000), and MMP2 (Proteintech, 10373-2-AP, dilution 1/1000), and the internal control was GAPDH (Abcam Biotechnology, Cambridge, UK, dilution 1/1000). Then incubated secondary antibody at 37°C for 30 min. The proteins were detected by an Odyssey fluorescence scanner (Li-Cor, Lincoln, NE).