Screening For Active Substances Of Anti Hepatoma And Lung Cancer Cells And Study On Their Action Mechanism

Objective The aim of this study was to screen out antitumor compounds from extracts of Periploca sepium and Panax ginseng(XJP-x and RSY-x)by MTT,and further to explore their action mechanism,so as to provide the lead compounds and to lay a theoretical basis for the development of new antitumor drugs.Methods The most appropriate screening concentration was obtained with detecting the cytotoxicity of different concentrations of cis-Dichlorodiammineplatinum(DDP)on human hepatoma HEPG2 cells and lung cancer A549 cells with MTT,and calculating their inhibition rate.The cytotoxicities of XJP and RSY series of samples on HEPG2 and A549 were detected with MTT.Two compounds with strong cytotoxicity were screened out.Determined their structures.The apoptosis of HEPG2 and A549 induced by XJP-12 and RSY-28 was tested.The effects of XJP-12 and RSY-28 on TNF-a and Caspase-8 activities of HEPG2 and A549 were detected with human TNF-a Elisa kit and Caspase-8 activity assay kit.Results The IC50 of DDP were(15.81 ± 1.24)mg/mL,(24.67±2.12)mg/mL on HEPG2 and A549 respectively.When the concentration of DDP was 60mg/mL,the inhibition rate on the two cells were more than 75%,so the concentration could be used as an initial concentration for the screening.It was found that XJP-12,RSY-11 and RSY-28 had inhibition on HEPG2,XJP-12,RSY-25,RSY-28 and RSY-30 had inhibition on A549.XJP-12 and RSY-28 are pure compounds.Their structures are ICso of XJP-12 and RSY-28 on A549 are 18.7mM and 13.9mM respectively.Their IC50 on HEPG2 are 13.5mM and 12.1mM respectively.IC50 of DDP on the A549 and HEPG2 are 52.6mM and 82.2mM respectively.It was found that XJP-12 and RSY-28 could induce cell apoptosis with Hoechst staining.It was associated with the two compounds concentrations.XJP-12 and RSY-28 could increase content of TNF-a and Caspase-8 activity in cell,and there was a certain dose-effect relationship.Conclusions DDP could effectively inhibit the proliferation of HEPG2 and A549,and there was a certain dose-effect relationship.So it can be used as positive control.Two pure compounds,XJP-12 and RSY-28,witih anti-tumor activity on HEPG2 and A549 had been screened out from Periploca sepium and Panax ginseng.There is no report of the research on their anti-tumor.Their cytotoxicities on HEPG2 and A549 are stronger than those of DDP.They could induce tumor cell apoptosis by increasing content of TNF-? and Caspase-8 activity in tumor cell,so it could be accomplished by the TNFR1/TNF-? exogenous death receptor-mediated signaling pathway.