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目的探讨苦参碱对恶性黑素瘤细胞A375转移能力的影响及其作用机制。方法采用MTT法和膜联蛋白-V-FITC/PI双染色法分别检测不同浓度苦参碱对A375细胞增殖和凋亡的影响,半定量RT-PCR分析经苦参碱干预后A375细胞乙酰肝素酶mRNA的表达变化,细胞黏附实验检测细胞黏附能力的改变,基质胶侵袭实验观察细胞侵袭能力的变化。结果苦参碱浓度≥0．5mg/mL时可抑制A375细胞增殖并诱导细胞凋亡,其作用呈剂量依赖性；苦参碱浓度在0．125～0．5mg/mL时,能明显下调A375细胞乙酰肝素酶mRNA的表达,并能抑制细胞的黏附、侵袭能力(P＜0．01),其抑制效应呈剂量依赖性,在不同浓度组间比较差异均有统计学意义(P＜0．01)。结论苦参碱在体外能抑制A375细胞的黏附、侵袭能力。苦参碱抗肿瘤侵袭可能与抑制肿瘤细胞增殖、诱导细胞凋亡以及下调乙酰肝素酶的表达有关。
Objective To investigate the effect of matrine on metastasis of malignant melanoma cell A375 and its mechanism of action. Methods The effects of different concentrations of matrine on proliferation and apoptosis of A375 cells were detected by MTT assay and annexin-V-FITC/PI double staining method. Semi-quantitative RT-PCR analysis of maturation of A375 cells after matrine treatment. The expression of mRNA and protein was detected by cell adhesion assay. The changes of cell invasive ability were observed by matrigel invasion assay. Results The matrine concentration of ≥ 0.5 mg/mL inhibited the proliferation of A375 cells and induced apoptosis in a dose-dependent manner. When the concentration of matrine was 0.125 to 0.5 mg/mL, the A375 could be significantly downregulated. The expression of heparanase mRNA can inhibit the cell adhesion and invasion ability (P<0.01). The inhibitory effect is in a dose-dependent manner, and there is significant difference between different concentrations (P<0). .01). Conclusion Matrine can inhibit the adhesion and invasion of A375 cells in vitro. Anti-tumor invasion of matrine may be related to inhibition of tumor cell proliferation, induction of apoptosis and down-regulation of heparanase expression.