Effects and mechanism of luteolin on proliferation,apoptosis,migration and glycolysis of human hepatocellular carcinoma cell lines
Objective To observe the effects of luteolin on the proliferation,apoptosis,migration and glycolysis of human hepatocellular carcinoma cell lines and to explore their mechanism.Methods HepG2 and Huh7 cells cultured in vitro were divided into two groups.Cells in the control group were treated with DMEM medium,and cells in the experimen-tal groups were treated with 20,40,60,80 and 100 µmol/L luteolin,respectively.Cell proliferation rate was measured by CCK-8 assay.The number of cell clones formed was detected by the cell clone formation experiment to evaluate the cell proliferation ability.The relative expression levels of apoptosis-related proteins(Bax,Bcl-2,and Caspase-3 proteins)were measured by Western blotting to evaluate the apoptosis ability.The cell migration rate was measured by the cell scratch test to evaluate the cell migration ability.The glucose consumption and lactic acid production levels were deter-mined by the kit.Glycolysis-related proteins(GLUT4,PDK1,HK2,PKM2,and LDHA proteins)were detected by West-ern blotting to evaluate the cell glycolysis ability.Western blotting was used to detect the LKB1/AMPK signaling pathway-related proteins(LKB1/P-LKB1 and AMPK/P-AMPK protein ratios).Results Compared with the control group,the proliferation rates of HepG2 and Huh7 cells in the experimental group decreased in a time-and dose-dependent manner;the number of colony-forming cell colonies of HepG2 and Huh7 cells in the experimental group decreased in a dose-depen-dent manner(all P<0.05).Compared with the control group,the relative expression levels of Bax and Caspase-3 proteins in HepG2 and Huh7 cells in the experimental group increased,and the relative expression of Bcl-2 protein decreased in a dose-dependent manner(all P<0.05).Compared with the control group,the migration rates of HepG2 and Huh7 cells in the experimental group decreased in a time-and dose-dependent manner(both P<0.05).Compared with the control group,the glucose consumption of HepG2 and Huh7 cells in the experimental group(except for the dose of 20 µmol/L)decreased,and the lactate production increased in a dose-dependent manner;the relative expression levels of GLUT4,PDK1,HK2,PKM2 and LDHA proteins in HepG2 and Huh7 cells in the experimental group decreased in a dose-depen-dent manner(all P<0.05).Compared with the control group,the protein expression ratios of LKB1/P-LKB1 and AMPK/P-AMPK in the experimental group increased in a dose-dependent manner(both P<0.05).Conclusions Luteolin inhib-its the glycolysis process of human hepatocellular carcinoma cell lines,further inhibits cell proliferation,migration and promotes apoptosis in a dose-or time-dependent manner,and the mechanism may be related to the activation of intracellu-lar LKB1/AMPK signaling pathway.
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