熊果酸诱导人肝癌细胞株HepG2凋亡的分子机制研究  

Molecular mechanism of ursolic acid induced apoptosis of human Hep G2 cells

作　　者：周力[1] 

机构地区：[1]中南大学肝胆肠研究中心,湖南长沙410008

出　　处：《中国现代医学杂志》2015年第5期24-28,共5页China Journal of Modern Medicine

摘　　要：目的探讨熊果酸对人肝癌Hep G2细胞增殖及凋亡的影响及其作用机制。方法常规培养肝癌细胞系Hep G2,进行细胞传代及细胞冻存;CCK-8法检测不同浓度熊果酸对Hep G2细胞增殖的影响;流式细胞术检测熊果酸处理后Hep G2细胞的凋亡率;Western blot检测运用熊果酸处理48 h后p-Akt及Bcl-2蛋白表达变化;实时定量PCR检测熊果酸处理后PTEN m RNA表达。结果细胞增殖抑制试验显示熊果酸呈浓度依赖性抑制Hep G2细胞生长;0、20和40μmol/L药物处理细胞48 h后其早期凋亡率分别为(2.04±0.13)、(13.41±1.72)和(16.58±2.45)%,晚期凋亡率分别为(3.17±0.42)、(18.36±2.53)和(23.59±1.88)%,差异有统计学意义(P<0.05);与对照组相比,熊果酸作用的Hep G2细胞p-Akt及Bcl-2蛋白表达水平均降低(P<0.05),PTEN m RNA表达水平上升(P<0.05)。结论熊果酸能够浓度依赖地抑制Hep G2细胞增殖,其机制可能是通过抑制PTEN/P13K/Akt信号转导通路来实现的。【Objective】 To investigate the effect of astragalus ursolic acid on proliferation and apoptosis of human hepatocellular carcinoma cell line Hep G2 and the underlying mechanisms. 【Methods】 Human hepatocellular carcinoma cell line Hep G2 was cultured under 37℃ and 5% CO2, and appropriate cell passage and cryopreservation were performed. After treatment for 48 h with different concentrations of ursolic acid, the proliferative activity and apoptosis rate of Hep G2 cells were examined by CCK-8 assay and flow cytometry, respectively. The expressions of p-Akt and Bcl-2 proteins in ursolic acid-treated Hep G2 cells were evaluated by Western blot. The expression of PTEN m RNA was detected by quantified real-time PCR. 【Results】 The proliferative activity of Hep G2 cells was obviously suppressed by ursolic acid treatment,and the inhibition rate increased in a dose-dependent manner. The flow cytometry showed that, compared with the control group, the apoptosis rates of Hep G2 cells were significantly increased after treatment with ursolic acid for 48 h. The early apoptosis rates were（2.04 ± 0.13）%,（13.41 ± 1.72）% and（16.58 ± 2.45）%, and the late apoptotic rates were（3.17 ± 0.42）%,（18.36 ± 2.53）% and（23.59 ± 1.88）%, the differences were statistically significant（P 〈0.05）.Western blot showed that the expressions of p-Akt and Bcl-2 proteins in Hep G2 cells decreased significantly after treatment with ursolic acid（P 〈0.05）. In contrast, the m RNA level of PTEN increased（P 〈0.05）, which was shown by real-time PCR. 【Conclusion】 Ursolic acid could repress cellular proliferation activity of Hep G2 cells, which could be attributed to inhibition of PTEN/ P13K/Akt signaling pathway.

关 键 词：肝癌 熊果酸 PTEN AKT 

分 类 号：R979.1[医药卫生—药品]