大蒜辣素致肝癌细胞HepG2凋亡探究.docVIP

大蒜辣素致肝癌细胞HepG2凋亡探究 　作者：燕丹 王坚 袁燿佐 陈志强 【摘要】 目的研究大蒜辣素（Allicin）导致肝癌细胞HepG2凋亡的现象及其影响规律。方法运用CCK－8试剂盒研究Allicin对HepG2细胞增殖的影响，运用流式细胞仪检测其对细胞凋亡、DNA代谢的影响，运用免疫印迹（Western blot）研究肝癌细胞凋亡基因表达的影响。结果Allicin能抑制HepG2细胞的增殖，细胞倍增时间延长为4 d，同时Allicin也能导致细胞出现大量凋亡，实验结果表明其凋亡率最高为4.13％，死亡率达到70.38%。细胞周期实验结果表明Allicin能使大部分细胞处于DNA合成期的S期及G0/G1期，延缓细胞增殖。Western blot实验结果表明Allicin能激活细胞凋亡基因提高药物的治疗效果。结论Allicin能抑制肝癌细胞的增殖可导致细胞大量死亡并引起细胞凋亡，并能引起细胞DNA代谢发生紊乱，细胞大部分被阻滞在DNA合成的G0/G1期，细胞分裂被抑制，Western blot实验结果表明细胞凋亡基因大量表达，细胞发生凋亡。 【关键词】 大蒜辣素； 生长曲线； 凋亡； DNA周期； HepG2肝癌细胞； Western blot； 免疫印迹 　　Abstract：ObjectiveTo investigate the effects of apoptosis on heptoma cell lines HepG2 caused by allicin. MethodsCCK-8 was exerted to investigate the proliferation of the HepG2 cell and cytometry machine was used to detect the apoptosis and mortality rate, finally western blot method was used to study the apoptosis protein expression. ResultsExperimental results showed that allicin could inhibit the proliferation of cells and cause the cell apoptosis and death, and the highest ratio of apoptosis and death were 4.13% and 70.38%, respectively. Cell cycle experimental results indicated that the DNA metabolism was disturbance and most of them were blocked in G0/G1 period resulting in cells mitosis were blocked too. Western blot results showed that allicin the actived apoptosis gene of cells.ConclusionAllicin can inhibit the HepG2 cell proliferation and cause cell death and apoptosis, furthermore Allicin can cause cell DNA metabolism disturbance.Western blot experiments results show that the apoptosis gene can increase expression. 　　Key words： Allicin; Proliferation curve; Apoptosis; Cell cycle; Western blot; HepG2 　　大蒜辣素(Allicin)是新鲜大蒜(garlic)有效成分提取物，分子式为C6H10OS2［1～3］。研究表明，Allicin可以降低血清胆固醇和甘油三酯水平，减少动脉粥样硬化的形成以及血小板的凝集，同时还具有抑制微生物的作用，可以抑制癌细胞增殖及诱导癌细胞的凋亡，并表现出了肿瘤杀灭作用。已有Allicin对癌细胞生物效应的研究主要集中在胃癌及肠癌等的研究［4］，对肝癌的研究甚少。本文即研究Allicin对人肝癌细胞HepG2的增殖及凋亡的影响，通过探讨细胞DNA代谢紊乱及凋亡基因表达进行分子学水平的机理探讨。 　　1 材料及试剂 　　UV2550型紫外分光光度计，Waters 2996高效液相色谱仪，Empower数据处理软件，细胞培养