大蒜素抗人肝癌HepG-2细胞的实验研究  

Research of Allicin's Anti-Cancer Effects on HepG-2 Cell Line

作　　者：陈传军[1] 于志坚[2] 吕亚莉[1] 蔡英[1] 

机构地区：[1]新昌县人民医院肿瘤内科,浙江新昌312500 [2]南通大学附属医院肿瘤中心,江苏南通226000

出　　处：《中华中医药学刊》2014年第7期1763-1767,I0013-I0015,共8页Chinese Archives of Traditional Chinese Medicine

摘　　要：目的:研究大蒜素对人肝癌细胞HepG-2生长的影响,探讨其抗人肝癌HepG-2细胞作用以及相关机制。方法:选用人肝癌HepG-2细胞为研究对象。通过倒置显微镜及hoechst染色荧光显微镜观察大蒜素作用后细胞形态的变化。MTT法检测不同浓度大蒜素作用后细胞增殖活性改变并计算抑制率(IR)。流式细胞术(FCM)检测大蒜素对细胞周期及凋亡率的影响。免疫细胞化学染色了解大蒜素作用后HepG-2细胞Bcl-2、Bax、Caspase-3表达的变化。结果:大蒜素作用24 h后人肝癌HepG-2细胞形态发生改变,表现为细胞缩小变圆,突起消失,分部稀疏,细胞核浓缩,边缘化,且随着大蒜素浓度增加,上述改变逐步明显。Hoechst染色荧光倒置显微镜观察对照组细胞核呈均匀一致的荧光,而实验组可观察到细胞凋亡形态变化,表现为细胞核固缩,染色质凝聚,边缘化,部分碎裂,呈致密的颗粒状强荧光。MTT法检测结果显示不同浓度的大蒜素(12.5μg/mL、25μg/mL、50μg/mL、100μg/mL)在不同时间(24 h、48 h、72 h)作用于HepG-2细胞后细胞增殖活性受到抑制,与阴性对照组及阳性对照组相比大部分组别差异有统计学意义。流式细胞仪检测结果提示与阴性对照组相比大蒜素组G0-G1期比例明显增加,细胞凋亡率明显上升。免疫细胞化学染色表明大蒜素作用后细胞Bax表达水平上升,Bcl-2/Bax比例下降,Caspase-3表达上升。结论:大蒜素对人肝癌HepG-2细胞的增殖有抑制作用,并能诱导HepG-2细胞凋亡。在一定范围内随着大蒜素浓度的增高,时间的延长,细胞增殖活性逐步降低,凋亡率逐步上升,呈时间-剂量依赖性。大蒜素抑制肝癌细胞增值诱导其凋亡与G0-G1期阻滞有关,经大蒜素作用后G0-G1期细胞比例明显高于对照组,其分子机制涉及上调Bax、Caspase-3表达。Objective：To investigate the anti-cancer effects of allicin on HepG-2 cell line by cell culture and try to find the related molecule mechanism of the effects.Method：HepG-2 cell line was taken as investigated subject,and the cell form＇s change was studied by invert microscope and hoechst stain invert fluorescence microscope after vary concentration allicin affected.MTT assay was used to study the cell＇s proliferative ability and calculate cell＇s inhibition ratio.The change of cell cycle and apoptosis rate were observed by flow cytometry.The change of the expression of Bcl-2,Bax and Caspase-3 were studied by immunocytochemical stain.Results：In test group,after 24 hours,HepG-2 cells ＇ form changed.The cells became more rounder than control group,and cells ＇ apophysis lost and distributing become very sparse.The nuclear concentrated and marginalized.The concentration of allicin were more higher and the changes became more clear.MTT assay demonstrated that the proliferation of HepG-2 cell can be inhibited by different concentrations allicin with different hours.Compared with negative control group and positive control group,the difference of the majority of test group had statistical significance.Flow cytometry showed that allicin group compared with negative group,the proportion of G0-G1 phase added and the cell apoptosis rate raised dramatically,and demonstrated the dose dependent manner.Immunocytochemical stain demonstrated that allicin can increase the expression of Bax and then the ratio of Bcl-2/Bax decreased and the expression of Caspase-3 was increased dramatically.Conclusion：Allicin can inhibit proliferation and induce apoptosis of HepG-2 cell in the certain scope.With the concentration of allicin raises,the proliferation was decreased and the apoptosis rate was increased,demonstrating the time-dose dependent manner.Allicin＇s effect of proliferation inhibition and apoptosis induction of HepG-2 cell is related to G0-G1 inhibition.The molecule mechanism of allicin＇s anti-cancer effects con

关 键 词：大蒜素 HEPG-2细胞 增殖 凋亡 BCL-2 BAX CASPASE-3 

分 类 号：R735.7[医药卫生—肿瘤]