To Explore The Mechanism Of Pulsatilla Saponins On Colorectal Cancer Cell Line HCT116 Based On PI3K/AKT/mTOR Signaling Pathway

Objective:To investigate the inhibitory effect of Pulsatilla saponins on human colon cancer cell line HCT116 and its preliminary mechanism.Method:Different concentrations of Pulsatilla saponins A3 and B4 were applied to the human colorectal cancer HCT116 cells cultured in vitro.The CCK-8 method was used to detect the effect on the proliferation of HCT116 cells.The qPCR were used to detect the effects of Pulsatilla saponins A3 and B4 on the target genes related to the PI3K/AKT/mTOR signal pathway.Results:1.Inhibition of Pulsatilla Saponin A3 and B4 on HCT116 Cells(1)Pulsatilla saponins A3 and B4 administration can exert drug effects,but the low concentration group has no obvious inhibitory effect,and the high concentration group has obvious inhibitory effect on proliferation.(2)Pulsatilla saponins A3 and B4 can exert drug effects after 24h and 48h,and the effect of high concentration group is time dependent.This effect is dose-dependent.2.Effects of Pulsatilla Saponin A3 and B4 on PI3K/AKT/mTOR signaling pathway Related Target Genes(1)Pulsatilla saponin A3 group CAV1 gene expression was significantly down-regulated(P<0.05),HIF1,PIK3CA,PTEN,MTOR,RHOA gene expression was significantly up-regulated(P<0.05),FIGF has an upward trend.For PIK3CA and PTEN target genes,Pulsatilla saponin A3 is stronger than B4.(2)Pulsatilla saponin B4 group CAV1,RHOA gene expression was significantly down-regulated(P<0.05),HIF1 had a downward trend,PTEN and MTOR gene expression was significantly up-regulated(P<0.05),PIK3CA,AKT1 had an upward trend.For the target genes of CAV1 and MTOR,the effect of Pulsatilla saponin B4 is stronger than that of A3.(3)Against the HIF1 and RHOA target genes,Pulsatilla saponins A3 and B4 have opposite effects.Conclusion:Pulsatilla saponins A3 and B4 can effectively inhibit the proliferation of HCT116 cells.Taken together,the effects of Pulsatilla saponins A3 and B4 are dose-dependent.The high concentration group is higher than the low concentration group.The antitumor effects of Pulsatilla saponins A3 and B4 are produced through the coordinated integration and regulation of multiple pathways and multiple targets,which may be related to the up-regulation of PTEN gene in PI3K/AKT/mTOR signaling pathway.