三七总皂苷通过调节Caspase-3、Caspase-9的表达诱导肝癌细胞BEL-7404凋亡的研究

A study on apoptosis of hepatocellular carcinoma BEL-7404 cells induced by panax notoginseng saponins by regulating the expression of Caspase-3 and Caspase-9

目的观察三七总皂苷(PNS)对肝癌细胞BEL-7404细胞线粒体膜电位、凋亡、Caspase-3、Caspase-9表达的影响,并探讨其诱导肝癌细胞BEL-7404凋亡的机制。方法将BEL-7404细胞分为:对照组、模型组、PNS组。对照组不加药物培养,模型组加阿霉素(10μg/m L)培养,PNS组用不同浓度PNS作用于BEL-7404细胞。CCK-8测定细胞抑制率、流式细胞术检测凋亡、线粒体膜电位,ELISA法检测Caspase-3、Caspase-9表达。结果不同浓度的PNS均可降低BEL-7404细胞线粒体膜电位,100μg/m L的PNS作用肝癌细胞48 h后,细胞线粒体膜电位较对照组降低(P<0.01),随着PNS浓度的增大,细胞凋亡率较对照组升高(P<0.01)。与对照组比较,PNS对BEL-7404细胞Caspase-3、Caspase-9的表达均有不同程度的增加。20μg/m L的PNS作用于BEL-7404细胞48 h后,Caspase-9的表达量较对照组上升(P<0.05); 40μg/m L的PNS作用于BEL-7404细胞48 h后Caspase-3的表达量较对照组上升(P<0.05)。结论一定剂量的三七总皂苷能降低BEL-7404细胞线粒体膜电位,可能是活化Caspase-9后激活凋亡因子caspase-3而导致细胞凋亡。

Objective To observe the effect of panax notoginseng saponins(PNS)on mitochondrial membrane potential,apoptosis as well as the expressions of Caspase-3 and Caspase-9 in hepatocellular carcinoma BEL-7404 cells,and to explore the mechanism of BEL-7404 apoptosis induced by PNS.Methods Bel-7404 cells were divided into control group,model group and PNS group.Drug culture was not applied to the control group,the model group was cultured with adriamycin(10μg/mL),and BEL-7404 cells were treated with different concentrations of PNS in the PNS group.And then,cell inhibition rate was measured by CCK-8,apoptosis and mitochondrial membrane potential were detected by flow cytometry,and the expressions of Caspase-3 and Caspase-9 were detected by ELISA.Results Different concentrations of PNS could decrease the mitochondrial membrane potential of BEL-7404 cells.After treating with 100μg/mL of PNS for 48 h,the mitochondrial membrane potential of hepatoma cells was lower than that of the control group(P<0.01),and with the increase of PNS concentration,the apoptosis rate was higher than that of the control group(P<0.01).Compared with the control group,PNS increased the expressions of Caspase-3 and Caspase-9 in BEL-7404 cells in varying degrees.After treating with 20μg/mL of PNS for 48 h,the expression of Caspase-9 in BEL-7404 cells increased while comparing with that of the control group(P<0.05).And after treating with 40μg/mL of PNS for 48 h,the expression of Caspase-3 in BEL-7404 cells increased while comparing with that of the control group(P<0.05).Conclusion A certain amount of PNS can reduce the mitochondrial membrane potential of BEL-7404 cells,which is possible that the activation of Caspase-9 leads to apoptosis by activating the apoptosis factor Caspase-3.