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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 observe the effects of Morus alba extracts on the proliferation and migration of human hepatoma HepG2 cells and the expression of matrix metalloproteinase-9 (MMP-9) in vitro in vitro. Inhibitory effect and molecular mechanism of HepG2 cells. Methods HepG2 cells were cultured, the serums of Morus alba at high, middle and low concentrations were respectively added to HepG2 cells. The flow cytometry was used to detect the effects of Morus alba extracts on HepG2 cells The effect of Morus alba extract on the migration ability of HepG2 cells was observed by cell scratch assay. The effect of Morus alba on the expression of MMP-9 protein in HepG2 cells was detected by Western blotting. The expression of MMP- (Reverse transcription-polymerase chain reaction, RT-PCR) method was used to detect the effect of Morus alba extract on the expression of MMP-9 mRNA in HepG2 cells. Results HepG2 cells in the medium concentration group of Morus alba extracts were significantly arrested in G1 phase compared with the control group (P <0.05). In medium concentration group, the migration area of HepG2 cells in migration medium was significantly less than that in the control group (P <0.001) (P <0.001) and MMP-9 mRNA expression in HepG2 cells (P <0.001). Conclusion Morus alba extract can effectively inhibit the proliferation and migration of HepG2 cells, and its mechanism may be related to the decrease of MMP-9 protein expression.