黄芩素、LY294002对人肝癌细胞系SMMC-7721细胞增殖及凋亡的影响  

Effects of baicalein and LY294002 on proliferation and apoptosis of human liver cancer cell line SMMC-7721

作　　者：汪枫 喻小兰 夏纪毅 王晓燕 唐利 曹勇 唐小平 夏国栋 

机构地区：[1]西南医科大学附属医院,四川泸州646000 [2]西南医科大学附属中医医院 [3]西南医科大学肿瘤研究所

出　　处：《山东医药》2016年第41期1-5,共5页Shandong Medical Journal

基　　金：四川省科技厅项目(14JC01383-LH53)

摘　　要：目的：探讨黄芩素和 PI3K 特异性抑制剂 LY294002对人肝癌细胞系 SMMC-7721细胞增殖及凋亡的影响。方法体外培养 SMMC-7721细胞，20μmol／L DMSO 及0、1、5、10、20μmol／L 黄芩素分别处理 SMMC-7721细胞，显微镜观察细胞数量变化；20μmol／L DMSO 及0、1、2、5、10、20、50、100、200和300μmol／L 黄芩素和0、1、2、5、10、20、30μmol／L 的 LY294002处理 SMMC-7721细胞24 h，CCK8法检测细胞增殖活性。将 SMMC-7721细胞随机分为4组，A 组加入0μmol／L 黄芩素；B 组加20μmol／L DMSO；C 组加20μmol／L 黄芩素；D 组加20μmol／L 黄芩素和10μmol／L LY294002，培养24 h，流式细胞术检测细胞周期并计算细胞凋亡比例、RT-PCR 法和 Western blotting法检测凋亡相关因子 Bcl-2、Bax mRNA 和蛋白表达及 p-AKT 活性。结果 SMMC-7721细胞数量随黄芩素／LY294002浓度增加而显著减少，细胞增殖活性明显降低（P 均＜0．05）。与 A、B 组相比，C、D 组的 G0～G1期细胞比例明显增多（P ＜0．05），S 期细胞比例明显下降，G2／M期细胞变化无统计学意义（P ＞0．05），C、D 组 G0～G1期细胞升高比例、S 期细胞降低比例比较，P ＜均0．05。与 A 组和 B 组相比，C、D 组早期凋亡和晚期凋亡细胞均明显增多（P 均＜0．05），D 组比 C 组早期凋亡和晚期凋亡细胞增多更明显（P ＜0．05）。与 A、B 组相比，C、D 组抑制凋亡因子 Bcl-2 mRNA 水平显著降低（P ＜0．05），凋亡促进因子 Bax mRNA 水平显著增高（P ＜0．05）；D 组 Bcl-2 mR-NA 水平比 C 组降低更明显，而 Bax mRNA 水平比 C 组升高更明显（P 均＜0．05）。与 A、B 组相比，C、D 组凋亡抑制因子 Bcl-2表达量和 AKT 磷酸化水平显著降低（P 均＜0．05），凋亡促进因子 Bax 表达量显著增高（P 均＜0．05），D组凋亡抑制因子 Bcl-2表达量和 AKT 磷酸化水平降低比 C 组更明显（P 均＜0．05），凋亡促进因子 Objective To investigate the effects of baicalein and phosphoinositide 3 kinase inhibitor LY294002 on proliferation and apoptosis of human liver cancer cell line SMMC-7721.Methods SMMC-7721 cells were cultured in vitro,and the cell proliferation of SMMC-7721 cells was measured by CCK8 when SMMC-7721 cells were treated with 20μmol/L DMSO and 0,1,5,10,20 μmol/L baicalein.The cell proliferative activity of SMMC-7721 cells was measured by CCK8 when SMMC-7721 cells were treated with 20 μmol/L DMSO,0,1,2,5,10,20,50,100,200 and 300 μmol/L baicalein and 0,1,2,5,10,20 and 30 μmol/L LY294002.Then SMMC-7721 cells were randomly divided into 4 groups：group A which was added with 0 μmol/L baicalein,group B with 20 μmol/L DMSO,group C with 20 μmol/L ba-icalein and group D with 20 μmol/L baicalein and 10 μmol/L LY294002 for 24 hours.The cell cycle and apoptosis of＆nbsp;SMMC-7721 cells were detected by flow cytometry,the mRNA and protein levels of apoptosis-related factors Bcl-2,Bax as well as pAKT activity was detected by RT-PCR and Western blotting.Results SMMC-7721 cells were dramatically de-creased with the increasing concentrations of baicalein and LY294002,with a dose-dependent manner,CCK8 assay indica-ted that both baicalein and LY294002 dramatically inhibited the proliferation of SMMC-7721 （all P 〈0.05）.Compared with groups A and B,the proportion of cells in G0-G1 phase of groups C and D was significantly higher （P 〈0.05）.The proportion of cells in S phase was decreased and the cell changes in G2 /Mphase was not statistically different （P 〉0.05）. In the groups C and D,the proportion of cells in G0-G1 phase increased,and the proportion of cells in S phase decreased （all P 〈0.05）.Compared with groups A and B,the early and late apoptotic cells in the groups C and D were significantly increased （all P 〈0.05）,and the early and late apoptotic cells in the group D were more than those in the group C （P 〈0.05）.Compared with groups A and B,the expression of Bcl 2 mRNA in the groups

关 键 词：肝癌 黄芩素 磷酸肌醇 3 激酶抑制剂 人肝癌细胞系 SMMC-7721 细胞 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]