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目的探讨猕猴桃根多糖(Actinidia chinensis Planch polysaccharid,ACPS)对人胃腺癌细胞(SGC-7901)增殖和凋亡的影响,及对SGC-7901细胞磷酸化p38(p-p38)蛋白表达的影响。方法采用CCK-8检测不同浓度ACPS对SGC-7901细胞的24、48、72 h的抑制作用;流式细胞技术检测各浓度ACPS作用48 h后SGC-7901细胞凋亡的发生率;Western blot法检测各浓度ACPS作用SGC-7901细胞后前体半胱氨酰天冬氨酸酶-9(pro-caspase-9)、聚腺苷二磷酸核糖聚合酶(PARP)和p-p38蛋白量的表达,以及p38特异性抑制剂预处理细胞后pro-caspase-9、PARP和p-p38蛋白量的表达。结果与对照组比较,1、2.5、5、10 mg/mL ACPS作用胃癌SGC-7901细胞后吸光度下降(P<0.05);同时药物剂量越高,作用时间越长,吸光度越低(P<0.01);24、48、72 h IC50分别为7.43、3.88、1.32 mg/mL;ACPS能下调SGC-7901细胞中pro-caspase-9蛋白的表达(P<0.01),增加PARP剪切蛋白的表达(P<0.01);进一步研究发现,ACPS处理SGC-7901细胞24 h后,p38的磷酸化水平升高(P<0.05),p38特异性抑制剂处理细胞2 h后能抑制p38磷酸化表达,并能抑制ACPS诱导的细胞凋亡。结论 ACPS具有抑制人胃癌SGC-7901细胞增殖,诱导其凋亡的作用;激活p38途径,进而激活caspase-9和PARP,最终导致细胞死亡,可能是其诱导胃癌细胞凋亡的分子机制之一。
Objective To investigate the effects of Actinidia chinensis Planch polysaccharid (ACPS) on proliferation and apoptosis of human gastric adenocarcinoma cell line SGC-7901 and its effect on phosphorylation of p38 (p-p38) protein in SGC-7901 cells. Methods CCK-8 was used to detect the inhibitory effect of ACPS on SGC-7901 cells at 24,48 and 72 h. Flow cytometry was used to detect the apoptosis rate of SGC-7901 cells treated with ACPS at various concentrations. Western blot The expression of pro-caspase-9, PARP and p-p38 protein in SGC-7901 cells treated with various concentrations of ACPS , And the expression of pro-caspase-9, PARP and p-p38 protein after cells pretreated with p38 specific inhibitor. Results Compared with the control group, the absorbance of 1, 2.5, 5, 10 mg / mL ACPS cells in SGC-7901 cells decreased (P <0.05) ); IC50 were 7.43,3.88,1.32 mg / mL at 24, 48 and 72 h respectively; ACPS could down-regulate the expression of pro-caspase-9 protein and increase the expression of PARP-cut protein in SGC-7901 cells (P <0.01). Further study found that p38 phosphorylation of SGC-7901 cells was increased after treated with ACPS for 24 h (P <0.05), p38 phosphorylation was inhibited by p38 specific inhibitor for 2 h Can inhibit ACPS-induced apoptosis. Conclusion ACPS can inhibit the proliferation and induce apoptosis of human gastric cancer SGC-7901 cells. Activation of p38 pathway, which in turn activates caspase-9 and PARP, eventually leads to cell death, which may be one of the molecular mechanisms that induce the apoptosis of gastric cancer cell line SGC-7901.