Study on the Effects of Citrusinol on the Proliferation ，Migration and the Expression of Skeleton-related Proteins of Human Hepatocellular Cells HepG 2 and Its Interaction Mode with Skeleton-related Proteins / 中国药房

ABSTRACT

OBJECTIVE：To study the effects of citrusinol on proliferation ，migration and the expression of skeleton-related proteins of human hepatocellular cells HepG 2，and to investigate its interaction mode with skeleton-related proteins. METHODS ： CCK-8 assay was used to detect the effects of different concentrations （12.5，25，50，100，200 μmol/L）of citrusinol on the proliferation of HepG 2 cells for 24 h. HepG 2 cells were divided into negative control group ，citrusinol low-concentration and high-concentration groups （50，100 μmol/L citrusinol）. After treated for 24 h，the migration of HepG 2 cells was detected by cell scratch test ；cell migration rate was calculated. mRNA and protein expression of F-actin ，β-tubulin and Ezrin in HepG 2 cells were determined by RT-PCR and Western blotting assay. Molecular docking software Schrodinger 2015 was used to analyze the interaction mode between citrusinol and above 3 kinds of proteins. RESULTS ：Citrusinol showed significant inhibition effect on the proliferation of HepG 2 cells （P＜0.05 or P＜0.01），in dose-dependent trend. Compared with negative control group ，cell migration， mRNA and protein expression levels of F-actin ， β-tubulin， Ezrin were decreased significantly in citrusinol low-concentration and high-concentration groups （P＜0.05 or P＜0.01）. Molecular docking results showed that the citrusinol could form hydrogen bond and hydrophobic bond with the above 3 skeleton-related proteins. CONCLUSIONS ：Citrusinol can inhibit the proliferation and migration of HepG 2 cells，the mechanism may be associated with the down-regulation of mRNA and protein expression of F-actin ，β-tubulin and Ezrin. The mode of its interaction with skeleton-related proteins may be the formation of hydrogen bond or hydrophobic bond.