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 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 C and D was significantly decreased (P <0.05),and the level of Bax mRNA was significantly increased (P <0.05).The level of Bcl-2 mRNA in the group D was significantly lower than that in the group C,while the level of Bax mRNA was significantly higher than that in the group C (all P <0.05).Compared with groups A and B,the expression of Bcl-2 and the phosphorylation of AKT in the groups C and D was significantly decreased (all P <0.05)and the expression of Bax was significantly increased (all P <0.05). The expression of Bcl-2 and the phosphorylation of AKT in group D was significantly higher than that in the group C (all P<0.05)and the expression of Bax was significantly higher than that in the group C (all P <0.05).Conclusion Baica-lein or LY294002 could induce apoptosis and inhibit the proliferation of SMMC7721 cells by regulating the expression of ap-optosis-related factors and increasing the level of activated AKT,and the synergistic effect was found between them.%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 Bax 表达量升高比 C 组更明显（P 均＜0．05）。结论黄芩素与 LY294002可能通过调控凋亡相关因子的表达及增加活化的 AKT水平从而诱导 SMMC-7721细胞凋亡，抑制其增殖，且二者有协同效应。
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