中药地锦草乙醇提取物体外抑制胃癌细胞增殖活性及机制研究

目的:探讨地锦草乙醇提取物(EEEH)对人胃癌(GC)细胞BGC823、MGC803、SGC7901及AGS增殖活性的抑制作用及机制。方法:采用蒸馏和冻干技术制备EEEH,用不同浓度的EEEH(62.5、125、250和500μg/m L)处理GC细胞,并设置溶剂对照组和空白对照组。采用MTS法检测胃癌细胞存活率;采用流式细胞术检测胃癌细胞凋亡率和周期分布;采用Western blot法检测胃癌细胞增殖、凋亡以及周期相关蛋白的表达水平。结果:MTS细胞增殖实验结果显示,与对照组相比,随着地锦草乙醇提取物浓度增加,GC细胞的增殖能力逐渐减弱,差异均有统计学意义(P<0.05);流式细胞术检测结果显示,62.5、125和250μg/m L的EEEH处理GC细胞MGC803和AGS 24 h后,与对照组相比,GC细胞的凋亡率显著增加,差异均有统计学意义(P<0.05),并且AGS细胞的细胞周期阻滞在G0/G1期(P均<0.05);Western blot检测结果显示,与对照组相比,经EEEH处理过的胃癌细胞中增殖相关蛋白PCNA以及抗凋亡相关蛋白BCL-2和BCL-XL的蛋白表达水平显著降低,促凋亡相关蛋白caspase-8和caspase-3出现明显的活性裂解片段,表明经EEEH处理后胃癌细胞凋亡活性增强,差异均有统计学意义(P<0.05),而caspase-9的蛋白表达水平并无明显变化。结论:EEEH能够抑制GC细胞增殖,其机制可能与EEEH诱导细胞凋亡和周期阻滞有关。

Inhibitory effects of ethanol extract from Euphorbia humifusa on gastric cancer cells in vitro

OBJECTIVE: To investigate the inhibitory effects of ethanol extract from Euphorbia humifusa(EEEH) on activities of BGC823,MGC803,SGC7901 and AGS in human gastric cancer (GC) cells. METHODS: The EEEH was prepared by distillation and freeze-drying technology. The GC cells were treated with different concentrations of EEEH (0,62.5,125,250 and 500 μg/mL) and the solvent control. The cell survival rates were detected by the MTS method,the apoptosis rates and cell cycle distributions were detected by flow cytometry,and the protein expression levels of cell proliferation,apoptosis and cycle were detected by Western blot method. RESULTS: After treatment with different concentrations of EEEH,the proliferation of GC cells was significantly inhibited compared with the control group and the difference was statistically significant (P < 0.05). The results of flow cytometry show that the apoptosis rates of treated AGS cells increased significantly and the cell cycle was arrested in G0/G1 phase. The differences were significant from that of the control (P < 0.05). The results of Western blot test show that the expression levels of proliferation-related protein PCNA,anti-apoptosis-related protein BCL-2 and BCL-XL in the treated cells were significantly lower than those in the control group,and apoptosis-related protein caspase-8 and caspase-3 showed obvious lytic fragments,indicating increased activities. The differences were statistically significant (P < 0.05). However,there were no significant changes in the expression levels and activities of caspase-9 protein. CONCLUSION: EEEH inhibited GC cell proliferation via the induction of apoptosis and cycle arrest.