左金方通过上调Bax/Bcl-2下调MMP诱导人胃癌细胞SGC-7901的凋亡

Zuojinfang induced the apoptosis of human gastric carcinoma cells SGC-7901 via increasing ratio of Bax/Bcl-2 and decreasing mitochondrial membrane potential

目的研究左金方诱导人胃癌细胞SGC-7901凋亡的作用机制。方法采用MTT比色法观察左金方对SGC-7901的生长抑制情况;流式细胞仪PI单染亚二倍体峰分析细胞凋亡率;Hoechst 33258染色观察细胞形态学变化;Western blot方法检测Bcl-2、Bax蛋白表达以及JC-1试剂盒测定细胞膜电位。结果左金方能抑制细胞活性,与时间-剂量呈依赖关系。0.1、0.5和1.0 mg·m L-1左金方作用于SGC-7901细胞48 h后,细胞凋亡率分别为(11.60±1.24)%、(21.80±0.28)%、(36.40±1.59)%,与对照组(2.66±1.33)%比较差异均有统计学意义(P<0.01);Hochest染色细胞出现核固缩状和颗粒状荧光等典型的凋亡学特征;左金方可降低SGC-7901细胞Bcl-2基因表达和增加Bax基因表达;左金方作用SGC-790l 4 h后,可以明显引起细胞MMP的下降;1.0 mg·m L-1左金方组的作用尤其明显。结论左金方通过上调Bax/Bcl-2比率,下调MMP诱导了人胃癌细胞SGC-7901的凋亡。

Objective To determine the mechanism of the apoptosis on human gastric carcinoma cells SGC-7901 induced by Zuojinfang. Methods Growth inhibition of the SGC-7901 treated by traditional decoction of Zuojinfang was analyzed by MTT assay. The apoptosis rates of the cells were determined by flow cytometry. The morphology was observed by Hoechst 33258 staining. The expressions of Bcl-2 and Bax proteins were measured with Western blot and the mitochondrial membrane potential was detected by JC-1 Kit. Results Zuojinfang inhibited the cell activity, and the cell survival rates decreased in a time-dose-dependent manner. When the cells were treated with 0.1, 0.5 and 1.0 mg·mL^- 1 traditional decoction of Zuojinfang for 48 h, the percentages of apoptosis were（11.60±1.24）%,（21.80±0.28）% and（36.40±1.59）%. The differences in comparison with the control group（2.66±1.33）% were significant（P〈0.01）. Hochest staining showed cell nuclear condensation, granular fluorescence and other typical features of apoptosis. Traditional decoctions of Zuojinfang reduced the Bcl-2 gene expression and increased gene expression of Bax. JC-1 Kit staining showed that mitochondrial membrane potential of SGC-7901 cells was decreased and the ratio of green to red fluorescence increased significantly after the incubation with traditional decoction of Zuojinfang for 4 h, and the experimental results was consistent with the testing results of apoptosis. The effect of 1.0 mg·mL^- 1 Zuojinfang group was particularly obvious. Conclusion Zuojinfang can induce the apoptosis of human gastric carcinoma cells SGC-7901 via increasing the ratio of Bax/Bcl-2 and decreasing the mitochondrial membrane potential.