土荆芥挥发油诱导人肝癌SMMC-7721细胞Caspase依赖性凋亡

Essential Oil of Chenopodium ambrosioides Induced Caspase-Dependent Apoptosis in SMMC-7721 Cells

目的:探讨土荆芥挥发油对人肝癌SMMC-7721细胞和人正常肝LO2细胞的抑制作用及体外抗肿瘤机制。方法:MTT法检测细胞增殖;碘化丙啶(PI)单染流式细胞仪分析细胞周期;显微镜下观察细胞形态变化及凋亡效应;Annexin V-PI双染流式细胞仪检测细胞凋亡率。结果:土荆芥挥发油对细胞增殖的抑制作用均呈浓度和时间依赖性(P<0.05),24、48、72 h处理其对SMMC-7721细胞的IC50均小于LO2细胞(P<0.05);土荆芥挥发油阻滞SMMC-7721细胞周期于G0/G1期;形态学观察表明细胞皱缩,粘附性降低;细胞核着橘红色,呈固缩状等晚期凋亡特征;细胞凋亡率呈浓度依赖性升高(P<0.05),加入Caspase抑制剂,土荆芥挥发油100μg/m L组晚期凋亡率显著降低(P<0.01)。结论:土荆芥挥发油能抑制人肝癌SMMC-7721细胞生长,其机制可能与阻滞细胞周期,诱导细胞发生Caspase依赖性凋亡有关。

Objective： To investigate the inhibition of cell proliferation by essential oil of Chenopodium ambrosioides on the human liver cancer SMMC-7721 cells and human liver LO2 cells,and to study the mechanism of anti-tumor in vitro. Methods： The inhibition of cell proliferation by essential oil of Chenopodium ambrosioides was determined by MTT assay; the distribution of cell cycle was analyzed by flow cytometry（ FCM） with PI staining; cell morphology and apoptosis effect of SMMC-7721 cells were observed by microscope; the apoptotic rate was quantified by FCM with Annexin V / PI double staining. Results： Essential oil of Chenopodium ambrosioides could significantly inhibit the cell proliferation in a concentration-time-dependent manner（ P〈0. 05）,and the IC50 values on SMMC-7721 cells were lower than human liver LO2 cells at 24,48 and 72 h,respectively（ P〈0. 05）; cell cycle of SMMC-7721 cells was arrested in G0/G1phase; morphological observation revealed that the cells were wrinkled and the cellular cohesiveness of cells was reduced; nuclear was condensed and in orange colour,of which were the late apoptotic features; and the apoptotic rate increased in a concentration-dependent manner（ P〈0. 05）,non-viable apoptotic rate was obviously decreased with caspase inhibitor in 100 μg / m L essential oil of Chenopodium ambrosioides（ P〈0. 01）. Conclusion： Essential oil of Chenopodium ambrosioides can inhibit SMMC-7721 cell proliferation,which may be related to inducing cell cycle arrest and caspase-dependent apoptosis.