Wang Yuan-Peng, Zhou Liang, Gong Xing-Guo
Institute of Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou 310058, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2013 Sep;42(5):504-10.
To investigate the effect of luteolin on cell growth and apoptosis of HepG2 cells in vitro.
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.
Luteolin effectively inhibited tumor cell proliferation in a dose-and time-dependent manner,and the inhibition rates of 20 μg/ml Luteolin for 72 h 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.
Luteolin has ant-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.
探讨木犀草素对体外培养的HepG2细胞生长和凋亡的影响。
用不同剂量（0 μg/ml、2.5 μg/ml、10 μg/ml和20 μg/ml）的木犀草素处理培养的HepG2、HL60、A549和LO2细胞，并设置不同时间（0 h、24 h、48 h和72 h）。采用MTT法检测细胞活力并计算IC50。用流式细胞术检测木犀草素处理6 h和12 h的HepG2细胞中的活性氧（ROS）水平。用流式细胞术和Annexin V-FITC/PI检测木犀草素处理24 h的HepG2细胞凋亡情况。用蛋白质印迹法检测HepG2细胞中凋亡通路蛋白（p53、ASPP2和iASPP）的表达水平，并分析剂量和时间效应。
木犀草素能有效抑制肿瘤细胞增殖，呈剂量和时间依赖性，20 μg/ml木犀草素作用72 h对LO2、HepG2、HL60和A549细胞的抑制率分别为39.34%、62.90%、57.57%和62.90%。木犀草素处理6 h和12 h后，HepG2细胞内ROS水平分别下降了13.88%和41.11%。木犀草素处理24 h的HepG2细胞凋亡率为14.43%，蛋白质印迹法显示，木犀草素处理12 h的HepG2细胞中，iASPP表达水平降低了77.07%，p53表达上调了179.37%，ASPP2表达上调了725.02%。
木犀草素对肝癌HepG2细胞具有抗增殖和促凋亡活性，这与HepG2细胞中促凋亡因子表达改变和ROS水平降低有关。
