三七总皂苷对人胰腺癌PANC-1细胞增殖、凋亡的影响

Effects of Panax Notoginseng Saponins on Proliferation and Apoptosis of Human Pancreatic Cancer PANC-1 Cells

目的研究三七总皂苷(PNS)对人胰腺癌PANC-1细胞增殖、凋亡的影响,为临床治疗胰腺癌提供新的化疗方案奠定理论基础。方法将培养的人胰腺癌PANC-1细胞分为三组,对照组(加PBS)、低浓度组(100μg·mL-1 PNS)、高浓度组(200μg·mL-1 PNS)。分别干预12、24、48 h后,CCK-8法检测人胰腺癌PANC-1细胞增殖活力;干预48 h后,流式细胞仪检测人胰腺癌PANC-1细胞凋亡情况,Hoechst 33258荧光染色观察细胞凋亡形态;Western blotting法检测人胰腺癌PANC-1细胞Bcl-2、Bax、Caspase-3、Caspase-9蛋白表达水平。结果PNS干预12、24、48 h后,低浓度组和高浓度组细胞增殖活力均显著低于对照组(P<0.05);PNS干预48 h后,低浓度组和高浓度组细胞凋亡数均显著高于对照组(P<0.05);低浓度组和高浓度组细胞Bcl-2水平显著低于对照组(P<0.05),Bax、Caspase-3、Caspase-9水平显著高于对照组(P<0.05)。结论PNS可抑制人胰腺癌PANC-1细胞的增殖活力,可能通过上调促凋亡蛋白Bax及下调抗凋亡蛋白Bcl-2的表达来激活Caspase级联反应,从而诱导PANC-1细胞凋亡。

Objective To study the effects of Panax notoginseng saponins(PNS)on proliferation and apoptosis of human pancreatic cancer PANC-1 cells,and to provide a theoretical basis for clinical treatment of pancreatic cancer.Methods Cultured human pancreatic cancer PANC-1 cells were divided into three groups:control group(with PBS),low dose group(100μg·mL-1),and high dose group(200μg·mL-1).After 12,24 and 48 hours of PNS administration,the proliferation of human pancreatic cancer PANC-1 cells was detected by CCK-8 assay.The effect of PNS on apoptosis of human pancreatic cancer PANC-1 cells was detected by flow cytometry after PNS was administered for 48 hours,and the apoptosis pattern of human pancreatic cancer PANC-1 cells was observed by Hoechst 33258 fluorescence staining.The effect of PNS on the expression of Bcl-2,Bax,caspase-3 and caspase-9 protein in human pancreatic cancer PANC-1 cells was detected by Western blotting.Results The CCK-8 assay showed that the cell proliferation vitality of the low-dose group and the high-dose group was significantly inhibited after 12 h,24 h and 48 h of PNS intervention(P<0.05).Flow cytometry showed after 48 hours of PNS intervention,the apoptosis rates of PANC-1 cells in the low-dose group and the high-dose group was significantly higher than in the control group(P<0.05).Western blotting showed that compared with the control group,the expression levels of Bcl-2 were lower in low-dose group and high-dose group than in control group,while the expression levels of Bax,caspase-3 and caspase-9 were higher in low-dose group and high-dose group than in control group(P<0.05).Conclusion PNS can inhibit the proliferation of human pancreatic cancer PANC-1 cells,possibly by up-regulating the expression of pro-apoptotic protein Bax,caspase-3 and caspase-9,and down-regulating the expression of anti-apoptotic protein Bcl-2.