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浙江大学学报（医学版）
浙江大学学报（医学版）  2013, Vol. 42 Issue (5): 504-510    DOI: 10.3785/j.issn.1008-9292.2013.05.005
论著     
木犀草素对人肝癌细胞HepG2的促凋亡作用
王远鹏，周亮，龚兴国
浙江大学生命科学学院生物化学所，浙江 杭州 310058
Pro-apoptotic effects of luteolin on hepatoma HepG2 cells
WANG Yuan-Peng, ZHOU Liang, GONG Xing-Guo
Institute of Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou 310058, China
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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。
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    
Abstract:

Objective： To investigate the effect of luteolin on cell growth and apoptosis of HepG2 cells in vitro.
Methods： Cultured HepG2，HL60，A549 and LO2 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 V-FITC/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.
Key words: Luteolin    Carcinoma, Hepatocellular    Liver neoplasms    Apoptosis    Reactive oxygen species    Tumor suppressor protein p53    Flow cytometry    Tumor cells, Cultured
收稿日期: 2012-09-14 出版日期: 2013-09-25
:  R 34  
基金资助:

国家自然科学基金（30900760）；浙江省科技计划项目（2006C12084）.
通讯作者: 龚兴国（1955-），男，研究员，主要从事重要药用蛋白和临床诊断用酶的基因工程研究；E-mail:gongxg@zju.edu.cn   
作者简介: 王远鹏（1986-），男，硕士研究生，主要从事信号传导通路和作用机制研究；E-mail:wangyuanpeng1986@yahoo.com.cn
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引用本文:

王远鹏, 周亮, 龚兴国. 木犀草素对人肝癌细胞HepG2的促凋亡作用[J]. 浙江大学学报（医学版）, 2013, 42(5): 504-510.

WANG Yuan-Peng, ZHOU Liang, GONG Xing-Guo. Pro-apoptotic effects of luteolin on hepatoma HepG2 cells. Journal of ZheJiang University(Medical Science), 2013, 42(5): 504-510.

链接本文:

https://www.zjujournals.com/med/CN/10.3785/j.issn.1008-9292.2013.05.005    或    https://www.zjujournals.com/med/CN/Y2013/V42/I5/504

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