摘要
[目的]探讨桔梗皂甙D(platycodin D,PD)对人胃癌细胞株SGC7901增殖的抑制作用及作用机制。[方法]体外培养人胃癌细胞株SGC7901,加入终浓度分别为5~20μm·L-1的PD。采用MTT法测定药物对细胞增殖的抑制作用,Annexin V/PI双标法检测细胞凋亡率,JC-1检测线粒体膜电位的变化,Western blot检测PD对凋亡相关蛋白cleaved caspase-3、cleaved caspase-9、cleaved PARP、bcl-2、bax和对MAPK信号通路蛋白ERK、JNK、p38及其磷酸化蛋白p-ERK、p-JNK、p-p38表达的影响。[结果]MTT结果显示,PD作用24h和48h后,PD对SGC7901细胞增殖的抑制随浓度的增加而增强;PD作用SGC7901后,细胞凋亡率明显增加,线粒体膜电位降低。Western blot检测结果显示cleaved PARP、cleaved caspase-3、cleaved caspase-9、bax蛋白表达量随着药物浓度的增加而升高,bcl-2蛋白表达下降,同时p-JNK、p-p38水平明显升高,p-ERK蛋白水平下降,ERK、JNK、p38蛋白的表达无明显变化。[结论]PD对胃癌细胞SGC7901有抑制增殖和诱导凋亡的作用。PD通过抑制ERK信号通路从而抑制细胞增殖,通过激活JNK和p38信号途径调控bax和bcl-2表达,导致线粒体膜电位下降,进而激活caspase,诱导癌细胞凋亡的发生。
[Objective] To investigate the effects of platycodin D(PD) on the proliferation and apoptosis of human stomach cancer SGC7901 and the related mechanism. [Methods] SGC7901 was cultured in virto and was treated with 5~20μm·L-1concentrations of PD. Cell proliferation was examined by MTT assay.Cell apoptosis was detected by Annexin V FITC/PI double staining. The change of mitochondrial trans-membrane potential was measured by JC-1 staining.The potein expression of cleaved caspase-3, cleaved caspase-9, cleaved PARP, bcl-2, bax, p-ERK, ERK, p-JNK, JNK, p-p38 and p38 detected by Western blot. [Results] MTT results showed that PD inhibited the growth of SGC7901 cells in a dose-dependent manner at 24 h and 48 h. SGC7901 cells treated with PD for 24 h showed significantly enhanced apoptosis and weakened mitochondrial membrane potential compared with the control cells. Western blot results showed that PD could up-regulate expression of cleaved PARP, cleaved caspase-3, cleaved caspase-9, bax, p-JNK, p-p38 protein, decreased bcl-2,p-ERK protein, the expression of ERK, JNK, p38 protein did not change significantly. [Conclusion] PD may inhibit the proliferation and induce the apoptosis of SGC7901 cells. These findings indicated that PD inhibited cell proliferation by inhibiting the ERK signaling. PD effect on bax and bcl-2 by activation of JNK and p38 signaling pathway resulted in the decrease of mitochondrial membrane potential and activation of caspase, which induced the apoptosis of cancer cells.
作者
代群
黄宣
陈喆
DAIQun HUANGXuan CHENZhe(First AffiliatedHospital of ghejiang Chinese Medical University, Hangzhou(310053)
出处
《浙江中医药大学学报》
CAS
2017年第7期573-579,共7页
Journal of Zhejiang Chinese Medical University
基金
国家自然科学基金项目(81673745)~~