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
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摘要
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( 10 μg/m L),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-3and Caspase-9 were detected by ELISA.Results Different concentrations of PNS could decrease the mitochondrial membrane potential of BEL-7404 cells.After treating with 100 μg/m L 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 20 μg/m L 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 40 μg/m L of PNS for48 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.
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( 10 μg/m L),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-3and Caspase-9 were detected by ELISA.Results Different concentrations of PNS could decrease the mitochondrial membrane potential of BEL-7404 cells.After treating with 100 μg/m L 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 20 μg/m L 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 40 μg/m L of PNS for48 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.
引文
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[2]林成,韦忠恒,黄瑞棋.斑蝥酸钠治疗原发性肝癌的机制及临床应用进展[J].右江医学,2017,45(6):744-747.
[3] Yang Y,Yuan P,Wei X,et al.Cultivated and wild Pleurotus ferulae ethanol extracts inhibit hepatocellular carcinoma cell growth via inducing endoplasmic reticulum stressand mitochondria-dependent apoptosis[J].Sci Rep,2018,8(1):13984.
[4]吴景华,郭佳培,曹青,等.青蒿素通过抑制IL-17/IL-17R表达诱导肝癌细胞凋亡的机制研究[J].现代预防医学,2017,44(4):697-700.
[5]强辉,冯敏,刘慧通,等.三七皂苷对激素性股骨头坏死早期细胞凋亡的影响及其机制[J].西安交通大学学报(医学版),2018,39(1):111-115.
[6] Rodenak-Kladniew B,Castro A,Strkel P,et al. Linalool induces cell cycle arrest and apoptosis in Hep G2 cells through oxidative stress generation and modulation of Ras/MAPK and Akt/m TOR pathways[J].Life Sci,2018,199:48-59.
[7]李响,李郑,程燚,等.三七总皂甙对人急性髓系白血病细胞株U937的抑制增殖及诱导凋亡作用[J].现代生物医学进展,2016,16(4):657-660.
[8] Zhang X,Chen Y,Cai G,et al.Carnosic acid induces apoptosis of hepatocellular carcinoma cells via ROS-mediated mitochondrial pathway[J]. Chem Biol Interact,2017,277(N):91-100.
[9] Shang N,Bank T,Ding X,et al.Caspase-3 suppresses diethylnitrosamine-induced hepatocyte death,compensatory proliferation and hepatocarcinogenesis through inhibiting p38 activation[J].Cell Death Dis,2018,9(5):558.
[10]焦佩娟,应小平.Caspase-3与肿瘤细胞凋亡关系的中医药研究进展[J].山东中医杂志,2017,36(8):721-724.