中草药单体石蒜碱和药根碱的抗黑色素瘤新生血管形成作用及其机制

发布时间：2018-08-02 18:31

【摘要】：黑色素瘤是一种严重危害人类健康的恶性肿瘤，近几年其发病率和死亡率都急剧上升，是肿瘤发病率增长最快的一种。但目前人们对于恶性黑色素瘤的发病机理尚不清楚，亦缺乏有效药物；已有的抗黑色素瘤中草药疗效不佳，需发掘新的更有效抗癌中草药。我们以肿瘤细胞为模型，从120多种中草药中筛选出两种抗黑色素瘤效果比较好的中草药石蒜和黄连，继而对石蒜的单体盐酸石蒜碱（LH）和黄连的单体盐酸药根碱（JH）的抗黑色素瘤血管形成作用，进行了深入研究。发现两者对人黑色素瘤C8161细胞主导的新生血管形成具有很好的抑制作用。盐酸石蒜碱（LH）对C8161细胞增殖的半数抑制浓度为0.99μM，能有效抑制C8161细胞主导的体内肿瘤新生血管的形成和体外类血管网状结构的形成；作用机制研究揭示，LH可以下调血管形成关键基因VE-cadherin在C8161细胞的表达，化学发光实验显示LH可明显抑制VE-cadherin启动子的活性。另外，LH可有效抑制C8161细胞增殖，使细胞周期阻滞于G0/G1期，而对细胞凋亡的作用不大；其机理与诱导细胞周期抑制物p21的高表达密切相关。此外，我们还研究了另一种中草药单体盐酸药根碱JH的抗黑色素瘤作用。JH对C8161细胞体外增殖的半数抑制浓度为47.35μM，，并能有效抑制C8161细胞主导的体内肿瘤新生血管的形成和体外类血管网状结构的形成，其机理与抑制VE-cadherin在C8161细胞的表达有关；另外，JH可使C8161细胞周期阻滞于G0/G1期，而对C8161细胞凋亡无明显影响，无明显毒副作用。综上所述，中草药单体盐酸石蒜碱和盐酸药根碱具有抗人高转移黑色素瘤C8161细胞主导的肿瘤新生血管形成的作用，其机制很可能是通过抑制VE-cadherin基因的转录，降低C8161细胞中VE-cadherin的表达水平；两者同时具有抑制C8161黑色素瘤生长和细胞增殖的作用。目前，国内外尚无LH和JH抗肿瘤血管形成研究的报道，本研究将为未来中医中药抗新生血管和黑色素瘤有效药物的研发提供新的信息和找到了新的突破口。
[Abstract]:Melanoma is a malignant tumor that seriously endangers human health. In recent years, the incidence and mortality of melanoma have increased rapidly. However, at present, the pathogenesis of malignant melanoma is not clear, and there is a lack of effective drugs, and the existing anti-melanoma Chinese herbal medicine is not effective, so it is necessary to explore new and more effective anti-cancer Chinese herbal medicine. Using tumor cells as a model, we selected two Chinese herbal medicines, Lycoris sativa and Coptis chinensis, which have a good anti-melanoma effect from more than 120 kinds of Chinese herbal medicines. The antiangiogenic effects of Lycorin (LH) and Rhizopine (JH) on melanoma were studied. It was found that both of them could inhibit the angiogenesis of human melanoma C8161 cells. The 50% inhibitory concentration of Lycorine (LH) on the proliferation of C8161 cells was 0.99 渭 m, which could effectively inhibit the formation of tumor angiogenesis and angioid reticular structure in vitro. The mechanism of LH could down-regulate the expression of angiogenesis key gene VE-cadherin in C8161 cells. Chemiluminescence assay showed that LH could significantly inhibit the activity of VE-cadherin promoter. In addition, LH could effectively inhibit the proliferation of C8161 cells and block the cell cycle in G0/G1 phase, but had little effect on cell apoptosis, and its mechanism was closely related to the overexpression of cell cycle inhibitor p21. In addition, We also studied the anti-melanoma effect of JH, another Chinese herbal monomer, and its inhibitory effect on the proliferation of C8161 cells in vitro. The median inhibitory concentration of JH on the proliferation of C8161 cells in vitro was 47.35 渭 m, and it could effectively inhibit the angiogenesis of tumor cells dominated by C8161 cells. The formation of vascular-like reticular structures in vitro, The mechanism is related to the inhibition of VE-cadherin expression in C8161 cells, in addition, JH can block the cell cycle in G0/G1 phase, but has no obvious effect on the apoptosis of C8161 cells. To sum up, Chinese herbal monomers Lycorine Hydrochloride and Rhizopine Hydrochloride have the effect of anti-tumor angiogenesis dominated by human high metastatic melanoma C8161 cells, and its mechanism is probably by inhibiting the transcription of VE-cadherin gene. Both of them can inhibit the growth and proliferation of C8161 melanoma. At present, there are no reports of LH and JH anti-tumor angiogenesis at home and abroad. This study will provide new information and find a new breakthrough for the research and development of anti-angiogenic and melanoma effective drugs of traditional Chinese medicine in the future.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.5

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