桑白皮提取物对HepG2细胞增殖和迁移的抑制作用

Inhibition of Cortex Mori on Cells Proliferation and Migration in HepG2

目的观察桑白皮提取物在体外实验条件下对人肝癌细胞HepG2细胞增殖、迁移及基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)表达的影响,探讨桑白皮提取物对人肝癌细胞HepG2细胞的抑制作用与分子机制。方法培养HepG2细胞,将高、中、低三种浓度的桑白皮提取物血清分别加入体外培养的肝癌细胞株HepG2细胞,应用流式细胞检测法检测桑白皮提取物对人肝癌细胞HepG2细胞周期的影响;应用细胞划痕实验观察桑白皮提取物对HepG2细胞迁移能力的影响;应用Western blotting检测桑白皮提取物对HepG2细胞MMP-9蛋白表达的影响;应用逆转录聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)方法检测桑白皮提取物对HepG2细胞MMP-9 mRNA表达的影响。结果桑白皮提取物血清中等浓度组HepG2细胞与对照组比较,明显阻滞于G1期(P<0.05);中等浓度组HepG2细胞,迁移实验中细胞迁移面积明显少于对照组(P<0.001);中等浓度组桑白皮提取物还可明显下调HepG2细胞MMP-9蛋白表达(P<0.001)及MMP-9 mRNA表达(P<0.001)。结论桑白皮提取物能有效抑制人肝癌细胞HepG2细胞增殖与迁移,其作用机制可能与降低MMP-9蛋白的表达有关。

Objective To investigate the inhibitory effect of Cortex Mori on proliferation migration and the expression of matrix metalloproteinase-9（MMP-9）in HepG2 cells in vitro and explore the molecular mechanism.Method Different concentrations of Cortex Mori serum were added into HepG2 medium respectively.Cell cycle assay was used to detect the effect of Cortex Mori serum on proliferation of HepG2 cells.Migratory ability was examined by using a model of wounding injury of confluent cultured cells.MMP-9 expression was detected by Western blotting.MMP-9 mRNA expression was detected by creverse transcription-polymerase chain reaction（RT-PCR）.Results The treatment of medium dose of Cortex Mori serum suppressed the proliferation of HepG2 cells,and blocked them at G1 stage.Compared with the control group,the migratory area was remarkably decreased（P〈0.001）.In addition showed that the expressions of MMP-9 at protein and mRNA level were significantly down-regulated（P〈0.001）.Conclusion The proliferative and migratory abilities of HepG2 cells could be inhibited by Cortex Mori serum,which may be related to down-regulating MMP-9 expressions.