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

WANG Feng; YU Xiaolan; XIA Jiyi; WANG Xiaoyan; TANG Li; CAO Yong; TANG Xiaoping; XIA Guodong

doi:10.3969/j.issn.1002-266X.2016.41.001

Abstract：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.