胃复春胶囊提取物对人胃癌细胞系MGC-803细胞凋亡及Bcl-2/Bax表达的影响

Weifuchun Capsule Extract Induces Apoptosis and Regulates the Expression Levels of Bcl-2/Bax in Human Gastric Cancer MGC-803 Cells

目的研究胃复春胶囊提取物对人胃癌细胞系MGC-803细胞的凋亡作用及其作用机理。方法MGC-803细胞分别用不同浓度的胃复春胶囊提取物处理。CCK-8法检测细胞活力;Hoechst 33258染色观察形态学改变;流式细胞仪检测细胞凋亡和细胞周期;Western blot检测Bcl-2、Bax、Caspase-3和Caspase-9的表达。结果胃复春胶囊提取物(浓度为0~100μg·mL^(-1))可以浓度和时间依赖性地降低MGC-803细胞的活力。Hoechst 33258染色显示胃复春胶囊提取物处理后的MGC-803细胞膜通透性增高。流式细胞检测结果显示胃复春胶囊提取物作用于MGC-803细胞后可以诱导细胞凋亡,细胞凋亡率分别为(20.4±2.7)%(25μg·mL^(-1))、(41.3±6.2)%(50μg·mL^(-1))、(68.8±5.8)%(100μg·mL^(-1)),明显高于对照组凋亡率(3.8±1.6)%(0μg·mL^(-1))(P<0.05)。MGC-803细胞经过不同浓度胃复春胶囊提取物处理24 h后,处于G0/G1期细胞比例升高,而G2/M期和S期细胞比例下降;免疫印迹实验结果表明胃复春胶囊提取物(浓度为25,50,100μg·mL^(-1))处理组MGC-803细胞中cleaved Caspase-3、cleaved Caspase-9和Bax表达量升高,而Bcl-2表达量降低,且呈剂量依赖性。结论胃复春胶囊提取物能抑制MGC-803细胞的增殖并诱导其凋亡,其作用机制与下调Bcl-2基因的表达,上调Bax基因的表达以及增加Caspases活性有关。

Objective To explore the effect of Weifuchun Capsule extract on cell apoptosis in human gastric cancer MGC-803 cells and the mechanism.Methods MGC-803 cells were treated with Weifuchun Capsule extract in different concentration.Relative cell viabilities were determined by the CCK-8 method;Hoechst 33258 staining was used to observe the morphological changes of cells;apoptosis and cell cycle were analyzed by flow cytometry;the protein expression levels of Bcl-2,Bax,Caspase-3,and Caspase-9 were detected by Western blot.Results Weifuchun Capsule extract(0~100μg/mL)significantly inhibited the viabilities of MGC-803 cells in a dose-and time-dependent manner.Hoechst 33258 staining showed that the membrane permeability of Weifuchun Capsule extract treated MGC-803 cells increased.Flow cytometry results showed that Weifuchun Capsule extract could induce apoptosis in MGC-803 cells,the apoptotic ratios are(20.4±2.7)%(25μg/mL),(41.3±6.2)%(50μg/mL),(68.8±5.8)%(100μg/mL),significantly higher than that of control group(3.8±1.6)%(0μg/mL)(P<0.05).After different concentrations of Weifuchun Capsule extract treatment for 24 h,the proportion of MGC-803 cells in G0/G1 phase increased,while the proportion of the G2/M phase and S phase reduced.In 100μg/mL Weifuchun Capsule extract treatment group,the proportion of cells in G0/G1 phase increased from 46.52%to 73.54%(P<0.01).The data of Western blot showed that Weifuchun Capsule extract(25,50,100μg/mL)up-regulated the expression levels of cleaved Caspase-3,cleaved Caspase-9,and Bax,but down-regulated Bcl-2,in a dose-dependent manner.Conclusion Weifuchun Capsule extract could inhibit the proliferation of MGC-803 cells and promote apoptosis,which may be associated with the down regulation of Bcl-2 expression and up regulation of Bax expression,as well as the increase of relative activity of Caspases.