木犀草素对人肝癌细胞HepG2的促凋亡作用  

Pro-apoptotic effects of luteolin on hepatoma HepG2 cells

作　　者：王远鹏[1] 周亮[1] 龚兴国[1] 

机构地区：[1]浙江大学生命科学学院生物化学所,浙江杭州310058

出　　处：《浙江大学学报（医学版）》2013年第5期504-510,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然科学基金(30900760);浙江省科技计划项目(2006C12084)

摘　　要：目的:研究木犀草素(Luteolin)对人肝癌细胞HepG2的生长抑制与促凋亡作用机制。方法:用梯度浓度木犀草素作用于多种肿瘤细胞,以MTT法检测药物对细胞的生长抑制作用,以流式细胞术检测HepG2细胞内活性氧水平,以Annexin V-FITC/PI双染法利用流式细胞术检测HepG2细胞凋亡情况,以Western-blot检测HepG2细胞凋亡相关通路蛋白表达水平。结果:木犀草素对多种肿瘤细胞的生长具有浓度和时间依赖的抑制作用(72 h最高抑制率超过60%),并能有效降低HepG2细胞内活性氧水平(约降低41.11%)。木犀草素作用下HepG2细胞凋亡率可达14.43%左右。木犀草素能够降低HepG2细胞内iASPP蛋白表达水平并上调p53与ASPP2蛋白表达。结论:木犀草素能够抑制肿瘤细胞生长,降低HepG2细胞内活性氧水平,并调节细胞内相关凋亡通路蛋白水平,从而诱导HepG2细胞凋亡。Objective：To investigate the effect of luteolin on cell growth and apoptosis of HepG2 cells in vitro.Methods：Cultured HepG2,HL60,A549 and L02 cells were treated with luteolin for different doses （0 μg/ml,2.5 μg/ml,10 μg/ml and 20 μg/ml） and varied times （0 h,24 h,48 h and 72 h）.Cell viability was measured with MTT assay and IC50 was calculated.The reactive oxygen species （ROS） levels in HepG2 cells treated with luteolin for 6 h and 12 h were measured with flow cytometry.Cell apoptosis of HepG2 cells treated with luteolin for 24h was examined with flow cytometry and Annexin VFITC/PI.Expression levels of apoptosis pathway proteins （p53,ASPP2 and iASPP） in HepG2 cells were detected with western blot and the dose and time-effect was analyzed.Results：Luteolin effectively inhibited tumor cell proliferation in a dose-and time-dependent manner,and the inhibition rates of 20μg/ ml Luteolin for 72h were 39.34％,62.90％,57.57％ and 62.90％ to LO2,HepG2,HL60 and A549 cells,respectively.The intracellular ROS level was decreased in HepG2 cells by 13.88％ and 41.11％after being treated with luteolin for 6 h and 12 h,respectively.The apoptosis rate of HepG2 cells treated with luteolin for 24 h was 14.43％,and western blot showed that luteolin reduced the expression level of iASPP by 77.07％ and up-regulated the expression of p53 by 179.37％ and ASPP2 by 725.02％ in HepG2 cells treated with luteolin for 12 h.Conclusion：Luteolin has anti-proliferative and pro-apoptotic activity on hepatoma HepG2 cells,which is associated with the altered expression of pro-apoptotic factors and decreased ROS level in HepG2 cells.

关 键 词：木犀草素 癌 肝细胞 肝肿瘤 细胞凋亡 活性氧 肿瘤抑制蛋白质P53 流式细胞术 肿瘤细胞 培养的 

分 类 号：R285.5[医药卫生—中药学] R735.7[医药卫生—中医学]