Inhibition Effect Of Betulinic Acid On Proliferation And Migration Of Gastric Cancer BGC-823 Cells

Objective:Gastric cancer is one of the most common malignant neoplasms of the aerodigestive tract,with the leading morbidity and mortality rates in China.Betulinic acid(BA)is a widely-derived pentacyclic triterpenoid compound that has been reported to be a potential therapeutic agent with strong anti-inflammation,antioxidant and antitumor activity.The higher expression level of vasodilator-stimulated phosphoprotein(VASP)in gastric cancer cell line BGC-823 was correlated with the migration behavior.In this study,BGC-823 cells were pretreated with BA for detecting the inhibition effect on proliferation and migration of the cells,and the expression level of VASP and upstream signal molecules were observed.Methods:Gastric cancer cells BGC-823 were cultured in vitro and pretreated with BA.MTT assay was used to detect the effect of BA on cell proliferation in different concentrations and different times.The effect of BA on cell clone formation was detected by plate cloning assay.Wound healing and transwell assays were used to explore the cell migration ability respectively.The effects of BA on the VASP expression at protein and mRNA levels were examined by Western blot and qRT-PCR.Luciferase assay was used to detect the promoter activity of VASP.The protein expression level of VASP and p-p65 was detected in the cells transfected with pCMV-P65 plasmid under the condition of BA treatment or not.Results:BA inhibited BGC-823 cell proliferation at a concentration of 10,20,30,40,50,60?M for 12,24 or 48h,respectively,in a dose and time-dependent manner.40?M BA could significantly inhibit clone formation ability.20p?M BA could significantly inhibit the BGC-823 cell migration and reduce the protein and mRNA levels of VASP.Transfection of pCMV-p65 plasmid could significantly increase the transcriptional activity via binding to VASP promoter region.Additionally,overexpression p65 could significantly increase the protein expression level of VASP,and 20?M BA treatment could block the up-regulation of VASP.Conclusions:The proliferation and migration ability of BGC-823 cells could be inhibited by BA through suppressing the protein expression level of VASP with the reduction of transcriptional activity via the NF-?B signal pathway.