花生红衣原花青素通过Wnt/β-catenin信号通路下调survivin表达诱导胰腺癌细胞凋亡

目的 探讨花生红衣原花青素(PSP)通过下调survivin表达诱导胰腺癌细胞凋亡的分子机制。方法 体外常规培养胰腺癌细胞株,配制100、200、300、400、500、600 μg/mL不同质量浓度的PSP,CCK-8法检测并计算不同质量浓度PSP在72 h内对胰腺癌细胞的抑制率,选取IC50=450 μg/mL作为PSP处理组剂量,并设对照组。Western blotting法检测胰腺癌细胞GSK-3β和β-catenin磷酸化程度、survivin、caspase-3和caspase-7蛋白表达水平,RT-PCR法检测胰腺癌细胞survivin mRNA表达水平,Annexin V-FITC/PI法检测24、48、72 h时PSP对胰腺癌细胞凋亡率的影响。 结论 与对照组相比,PSP处理组胰腺癌细胞GSK-3β和β-catenin磷酸化程度于12 h达到最高(P＜0.01)。PSP处理组胰腺癌细胞survivin mRNA的表达量与对照组比较明显降低(P＜0.01)。与对照组比较,在PSP的作用下,胰腺癌细胞中survivin蛋白表达量在72 h时最低(P＜0.01),caspase-3和caspase-7表达量在72 h时最高(P＜0.01),胰腺癌细胞凋亡率随着时间延长越来越高,在72 h时达到51.59%(P＜0.01)。结论 PSP可通过Wnt/β-catenin信号通路下调survivin蛋白表达诱导胰腺癌细胞凋亡。

Peanut Skin Procyanidins reduce the expression of survivin and induce the apoptosis of pancreaticcancer cells through Wnt/ beta-catenin signaling pathway

Objective The present work was undertaken to explore the molecular mechanism for the apoptosis of pancreatic cancer cells induced by PSP through down regulating the expression of survivin via the Wnt/β-catenin signal pathway.Methods Pancreatic cancer cells were nurtured routinely at PSP concentrations of 100,200,300,400,500 and 600μg/mL.The cell counting kit-8(CCK-8) assay was used to detect and calculate the inhibitory rate of pancreatic cancer cells after 72 hours of culturing.The IC50 of 450μg/mL was chosen as the experimental group. Western blotting was adopted to detect the degree of GSK-3β and β-catenin phosphorylation and the protein expression levels of survivin, caspase-3 and caspase-7.RT-PCR was used to evaluate the expression level of survivin mRNA.Annexin V-FITC/PI was used to test inhibitory rate of pancreatic cancer cells after 24, 48 and 72 hours.Results The degree of GSK-3β and β-catenin phosphorylation reached the highest (P＜0.01) after 12h in the experiment group.The expression of survivin mRNA in the experiment group was significantly lower (P＜0.01) compared to that in the control group.Under the action of PSP,the expression of survivin protein was the lowest (P＜0.01) after 72 hours while the expression of caspase-3 and caspase-7 protein were the highest(P＜0.01)after 72 hours.The apoptosis rate of pancreatic cancer cells increased as the culture period was prolonged,which reached 51.59% (P<0.01) after 72 hours of culture.Conclusion PSP can exert a pro-apoptotic effect on pancreatic cancer cells by reducing the expression level of survivin through the Wnt/β-catenin signal pathway.