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Journal of ZheJiang University(Medical Science)  2013, Vol. 42 Issue (5): 504-510    DOI: 10.3785/j.issn.1008-9292.2013.05.005
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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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 wordsLuteolin      Carcinoma, Hepatocellular      Liver neoplasms      Apoptosis      Reactive oxygen species      Tumor suppressor protein p53      Flow cytometry      Tumor cells, Cultured     
Received: 14 September 2012      Published: 25 September 2013
CLC:  R 34  
Cite this article:

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.

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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。

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 
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