Experimental Study About Total Saponins Of Paris Polyphylla Var. Yunnanensis On Human Tongue Squamous Cell Carcinoma CAL-27 Cell Proliferation And Mechanism

Objective:To investigate the effect of total saponins of paris polyphylla var.yunnanensis on proliferation of human tongue squamous cell line CAL-27 and explore the relevant mechanisms.Methods:CCK-8 method was used to detect cell proliferation inhibition rates after treated with the different concentrations of total saponins of paris polyphylla var.yunnanensis in 24h and 48h.The apoptotic rates and cell cycle in each experimental group was measured by Flow cytometry.Immunocytochemistry was used to detect the expression of MIF,AKT,and P53 in CAL-27 cells after treated with the total saponins of paris polyphylla var.yunnanensis for 24h.Western blot assay was used to analysis the proteins expression levels of MIF,AKT,and P53 in CAL-27 cells by treated with different concentrations of the total saponins of paris polyphylla var.yunnanensis for 24h.Results:1.The CCK-8 method showed that in the range of 10～60μg·mL-1 the different concentrations of the total saponins of paris polyphylla var.yunnanensis had a concentration-dependent dose-dependent effect on CAL-27 cells(P<0.05),but there was no time effect(P>0.05);.2.FCM showed that between control group and low-dose group(1.95±0.18%)had no difference in apoptosis rates(P>0.05),nevertheless,there was a significant difference compared with the medium-dose group and high-dose group(P<0.01).3.Immunohistochemistry showed that there was no significant difference in MIF expression between the low-dose group and the control group,but there was a statistically significant difference in the MIF levels between the medium-dose group and high-dose group(P<0.05);The expression of AKT and P53 proteins in each experimental group had significantly decreased(P>0.05)and there were significant differences compared with the control group(P<0.01).4.Western blot analysis showed that the expression of MIF protein in each experimental group conspicuous decreased,and the difference was statistically significant(P<0.05),but the low-dose group(P>0.05);Compared with the control group,the expression levels of AKT and P53 proteins in each experimental group was decreased,which was dose-effect dependent,and the difference was statistically significant(P<0.05).Conclusions:Total saponins of Paris polyphylla var.yannanensis significantly inhibited the proliferation of CAL-27cells induced apoptosis and disturbed the cell cycle,and down-regulating the expression of MIF,AKT and P53.