目的研究苦参碱(matrine,Ma)抑制人肝癌HepG2细胞增殖及其对血管内皮生长因子(vascular endothelial growth factor,VEGF)、基质金属蛋白酶-9(matrix meta-lloproteinase-9,MMP-9)表达的影响,探讨其抗肝癌作用可能的分子机制。方法设Ma组和阴性对照组,Ma组设5个药物梯度,采用0.2 mg/ml、0.4 mg/ml、0.8 mg/ml、1.6mg/ml、3.2mg/ml浓度的苦参碱分别作用于对数生长期的HepG2细胞,MTT法检测苦参碱对细胞生长的抑制作用,Real-time PCR检测VEGF mRNA表达,蛋白质印迹法(Western Blot,WB)检测VEGF和MMP-9蛋白的表达。结果苦参碱能抑制人肝癌HepG2细胞增殖,呈时间和剂量依赖性。苦参碱能下调VEGF mRNA表达(P〈0.05)和VEGF、MMP-9蛋白的表达(P〈0.05)。结论苦参碱在体外能抑制肝癌细胞增殖,呈时间和剂量依赖性,其可能的分子机制是通过下调VEGF和MMP-9蛋白表达,抑制肝癌血管生成,进而起到抗肝癌作用。
Objective To investigate the anti-hepatocellular carcinoma molecule mechanism of matrine (Ma) by observing cell proliferation and expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase 9 (MMP-9) in human liver cancer HepG2 cell lines. Methods A Ma group and a negative control group were created in this study. The Ma group included 5 drug concentrations gradients. The HepG2 cell lines at exponential phase were cultured in vitro and treated with 0.2 mg/ml, 0. 4 mg/ml, 0. 8 mg/ml,1.6 mg/ml and 3.2 mg/ml matrine, respectively. HepG2 cell lines viability was measured by a MTT assay. Expressions of VEGF mRNA were measured by using a Real-time PCR. A Western Blot was applied to examine the protein levels of VEGF and MMP-9. Results Matrine significantly inhibited the proliferation ofhuman cancar HepG2 cell lines in a dose- and time-dependent manner. Matrine could down-regulate VEGFmRNA expression ( P 〈0.05) and the expressions of VEGF and MMP-9 proteins ( P 〈0.05). ConclusionMatrine showed satisfactory results in inhibiting the proliferation of HepG2 cell lines in vitro in a dose- andtime-dependent manner, which might be achieved by down-regulating the expression of VEGF and MMP-9 and by inhibiting the hepatocellular carcinoma vascular formation for anti-liver cancer.