The Anti-gastric Cancer Effect Of Lily Total Saponins In Vitro And Its Preliminary Mechanism

Purposes:Gastric cancer,both the incidence and mortality rate are second in China,seriously damages human health,which is a common malignant tumor of the digestive tract.Gastric cancer is characterized by multidrug resistance,distant metastasis and recurrence.In recent years,it has been reported that tonic traditional Chinese medicines and its component of total saponins have a good therapeutic effect on malignant tumors.However,the effect of total saponins from lily on gastric cancer is not clear.Therefore,this study aims to elucidate the potential effect of total saponins of from Lilium lancifolium bulbs（TSLL）on the proliferation,apoptosis,invasion and metastasis of gastric cancer cells in vitro and to explore its possible mechanism.Methods:CCK-8 method,cell plate cloning assay and EdU staining were used to detect the effect of different concentrations of TSLL on the growth and proliferation of human gastric cancer SGC-7901 and HGC-27 cells.YF488-Annexin V/PI double staining and Hoechst 33342 staining were applied to observe the effect of different concentrations of TSLL on the apoptotic morphology of SGC-7901 and HGC-27 cells.And scratch test,Transwell invasion test as well as cell adhesion test were used to measure the effects of different concentrations of TSLL on the migration,invasion and adhesion of SGC-7901 and HGC-27 cells,respectively.Moereover,western blot was used to detect the changes of proliferation,apoptosis and invasion and metastasis associated proteins in gastric cancer SGC-7901 and HGC-27 cells treatment with different concentrations of TSLL.Results:TSLL significantly inhibited gastric cancer SGC-7901,HGC-27 cells growth after 24,48 or 72 h incubation.The values of IC_50（24h）were about 291.6,230.5μg·mL^-1,IC_50（48h）were about 246.5,108.4μg·mL^-1,and IC_50（72h）were about 183.3,71.9μg·mL^-1,respectively.TSLL markedly reduced gastric cancer SGC-7901 and HGC-27 cells clone formation rate and DNA synthetic ratio.TSLL markedly reduced gastric cancer SGC-7901 and HGC-27 cells clone formation.TSLL could induce SGC-7901 and HGC-27 cells apoptosis and significantly increase the rate of apoptosis.TSLL suppressed SGC-7901 and HGC-27 cells scratch wound healing,invasion of artificial basement membrane and adhesion to fibronectin.The results of western blot detection showed that TSLL could down-regulate the expression of PCNA,Bcl-2 and MMP-2 proteins,up-regulate the expression of Bax and TIMP-1 protein,and promote the activation of Casp-3 in SGC-7901 and HGC-27 cells.The TSLL could also significantly increase the expression level of p21 in SGC-7901 cells.In addition,the TSLL had no obvious effect on the expression of EMT marker protein E-Cad and Vim.Conclusion:The TSLL can inhibit the proliferation,migration,invasion and adhesion of SGC-7901 and HGC-27 cells,as well as induce cancer cells apoptosis.The anti-proliferation effect of TSLL may be by inhibiting the synthesis of intracellular DNA through up-regulating the expression of p21 and down-regulating the expression of PCNA in gastric cancer cells,thus leading to cell cycle arrest.The TSLL can also inhibit gastric cancer cells proliferation via induction of apoptosis,which is related to the regulation of Bcl-2 and Bax proteins expression in the cells.The TSLL can also down-or up-regulate the expression levels of MMP-2 and TIMP-1 proteins,thereby reducing the degradation of ECM by tumor cells and inhibiting the cells invasion and metastasis.Furthermore,the inhibitory effect of TSLL on the proliferation,invasion and metastasis of undifferentiated gastric cancer HGC-27 cells and the apoptosis-induce effect are better than the effect on the moderately differentiated SGC-7901 cells,suggesting that the total saponins seem to play a better antitumor effect for tumors with high degree of malignancy.