Effect of MLN4924 on the invasion and migration of HepG2 cells and the related mechanisms

Abstract

Objective To investigate the effect of MLN4924 which is a kind of NEDD8-activating enzyme (NAE) inhibitor on the invasion and migration of liver cancer and the related mechanisms. Methods Lab tests involved separating HepG2, Hep3B and SMMC-7721 cells into individual observation groups and observing the before and after effect with the intervention of MLN4924. Cell counting kit-8 (CCK-8) method was used to detect the cellular activity and half maximal inhibitory concentration (IC50) values. The changes of apoptosis were detected by flow cytometry. Cell Scratch and Transwell test were used to detect the effect of MLN4924 on invasion and migration. Western blotting method was used to detect the changes of E-cadherin and Vimentin during epithelial-mesenchymal transition (EMT). Results By using CCK-8 method, all three hepatoma cell lines were inhabited and HepG2 control group showed a sensitivity level towards MLN4924 with IC50 of 3.34 μmol/L. Flow Cytometry indicated that MLN4924 could significantly accelerate HepG2 apoptosis. Cell Scratch and Transwell test also confirmed MLN4924 had the potential to suppress the invasion and migration of HepG2 in a dose-dependent manner (P=0.001). Additionally, the relative expression of E-cadherin treated with 0.5, 1.0, 2.0, 4.0 μmol/L MLN4924 after 24 hours were 0.313±0.005, 0.594±0.016, 0.431±0.002, 0.780±0.019, 1.000±0.015, respectively, and the relative expression of Vimentin were 1.000±0.012, 1.158±0.035, 1.087±0.076, 0.635±0.028, 0.351±0.010. Conclusion MLN4924 showed positive results in inhibiting the growth of HepG2 accelerating the apoptosis, and also suppressing the invasion and migration of HepG2 through EMT. Key words: MLN4924; Hepatoma; Cell migration; Epithelial-mesenchymal transition