反左金丸水提物对人胃癌细胞SGC-7901凋亡及bcl-2、bax基因表达的影响

目的:观察反左金丸水提物(waterextractsofRetro-zuojinPill,WERZP)对人胃癌细胞(SGC-7901)生长的抑制和凋亡诱导效果,及其对bcl-2和bax表达的影响。方法:采用MTT比色法观察不同浓度的WERZP对SGC-7901的生长抑制情况;流式细胞仪检测细胞凋亡率;Hoechst33258染色观察细胞形态学变化以及Westernblot方法检测WERZP对人胃癌细胞SGC-7901中bcl-2、bax蛋白表达的影响。结果:WERZP作用SGC-7901细胞24h后,细胞存活率显著降低(P<0.05),且与时间、剂量呈依赖关系。流式细胞仪PI单染亚二倍体峰分析,0.1,0.5,1,5和10mg·mL^-1WERZP作用SGC-7901细胞24h,细胞凋亡率分别为(4.02±1.33),(36.10±0.23),(55.91±0.95),(56.50±1.55)和(60.27±3.58)%,其中(0.5～10)mg.mL^-1组与对照组(2.66±1.33)%比较差异均有统计学意义(P<0.01)。Hochest染色结果显示细胞出现核固缩状和颗粒状荧光等典型的凋亡学特征。WERZP可降低SGC-7901细胞bcl-2基因表达和增加bax基因表达。结论:WERZP对SGC-7901生长有明显的抑制作用并可诱导SGC-7901的凋亡,bcl-2基因表达降低和bax基因表达增加可能是其凋亡机制之一。

Effect of Retro-Zuojin Pill on Apoptosis and Bcl-2,Bax Gene Expression in SGC-7901 Cells

Objective:To investigate the effect of water extracts of Retro-Zuojin Pill(WERZP) on proliferation and apoptosis of human gastric carcinoma cells (SGC-7901),and the relationship between apoptosis and bcl-2,bax expression.Methods: The cell viability was measured by MTT;the apoptotic rate was investigated by FCM and the morphology was observed by Hoechst 33258 staining.Detection of bcl-2,bax gene and protein expression of WERZP-treated gastric cancer cells were carried out with Western blot method.Results: After treatment with WERZP for 24 h,the proliferation of SGC-7901 cells was significantly inhibited(P<0.05).Inhibited cell proliferation and induced cell death were in a dose-dependent and time-dependent manner,with apoptotic rate of(4.02±1.33),(36.10±0.23),(55.91±0.95),(56.50±1.55) and(60.27±3.58) % accordingly.All the concentrations except the lowest induced significant apoptosis compared with the control(P<0.01).The nucleuses were positively stained with fluorochrome Hoechst 33258.WERZP could reduce the bcl-2 gene expression and increase gene expression of bax.Conclusion: WERZP could significantly inhibit the growth of SGC-7901 cells and induce apoptosis,reduce the bcl-2 gene expression.The increased gene expression of bax may be involved in apoptosis mechanism.