木鳖子醇提物对黑素瘤B16细胞增殖的抑制及其可能机制

目的：观察木鳖子醇提物(ethanol extract from cochinchina momordica seed, CMSEE)对黑素瘤B16细胞增殖的影响，初步探讨其作用机制。方法：MTT法和克隆集落形成实验检测CMSEE对小鼠黑素瘤B16细胞生长的影响，倒置相差显微镜和瑞氏吉姆萨染色观察细胞形态学变化，流式细胞技术（FCM）检测B16细胞周期分布和凋亡率的变化，比色法检测CMSEE对B16细胞黑素生成和酪氨酸酶活性的影响，RTPCR法检测CMSEE对B16细胞中Cmyc、〖STBX〗P38和Tyr〖STBZ〗基因表达的影响。结果：CMSEE（10～100 mg/L）明显抑制B16细胞的增殖(P<0.05, P<0.01)，呈质量浓度和时间依赖性。10～40 mg/L CMSEE处理后，B16细胞呈现典型的细胞分化形态，细胞周期阻滞于G0/G1期，黑素产生和酪氨酸酶活性增加(P<0.01)，Cmyc基因表达下调，〖STBX〗P38和Tyr〖STBZ〗基因表达上调；100 mg/L CMSEE处理后的B16细胞呈现凋亡形态变化[凋亡率达(37.2±329)%]，黑素生成减少(P<0.05)，酪氨酸酶活性降低(P<0.01)。结论：CMSEE体外对B16细胞增殖有明显的抑制作用，其机制与其低剂量诱导分化和高剂量诱导凋亡有关。

Inhibitory effect of cochinchina momordica seed ethanol extract on proliferation of melanoma B16 cells and its possible meachanism

Objective:To investigate the effect of cochinchina momordica seed ethanol extract (CMSEE) on the proliferation of melanoma B16 cells and the underlying mechanism.Methods:MTT and clone formation assay were used to assess the effect of CMSEE on the growth of B16 cells.Morphological changes of B16 cells were observed under phase-contrast microscope and Giemsa staining.Cell cycle and apoptosis rate were examined by flow cytometry (FCM).The effect of CMSEE on melanin production and tyrosinase activity of B16 cel...