灯盏花素联合顺铂治疗小鼠黑色素瘤作用研究

Effect of breviscapine combined of cisplatin on malignant melanoma in mice

目的研究灯盏花素(Bre)、顺铂(Cis)以及两药联用对小鼠黑色素瘤的作用及其机制。方法用C57小鼠建立皮下移植瘤模型,小鼠分为4组,对照组(注射二甲基亚砜),Bre组(注射4μg·g^(-1)Bre),Cis组(注射5μg·g^(-1)Cis),Bre+Cis组(注射Bre与Cis),隔天注射,共计15 d,每次治疗后测量肿瘤体积。用免疫组织化学方法检测Brd U及CD31表达,流式细胞术检测肿瘤周期。用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法检测小鼠黑色素瘤细胞增殖。结果与对照组相比,Bre组、Cis组以及Bre+Cis组均能明显减小肿瘤体积和肿瘤重量(P<0.05,或P<0.01);同时,3组均能明显抑制黑色素瘤细胞的增殖与血管新生(P<0.05,或P<0.01);Bre组、Cis组以及Bre+Cis组对黑色素瘤的细胞凋亡率分别为(13.78±1.66)%,(12.90±1.10)%,(18.74±1.22)%,与对照组(2.02±0.32)%相比差异有统计学意义(P<0.01)。结论 Bre、Cis通过影响细胞周期以及血管形成,从而抑制小鼠黑色素瘤的生长。

Objective To explore the inhibitory effect of breviscapine（ Bre）,cisplatin（ Cis）,as well as the two combination on malignant melanoma in mouse and its mechanism. Methods Subcutaneous transplantation tumor model is established using C57 mice which were divided into 4 groups,control group treated with dimethyl sulfoxide（ DMSO）,Cis group treated with cisplatin of 5 μg·g^-1,Bre group treated with breviscapine of 4 μg·g^-1 and Cis ＋ Bis group treated with cisplatin and breviscapine. The tumor volume and weight was measured after each treatment. The change of tumor cells and tumor markers were observed by immunohistochemistry. Cell cycle was detected by flow cytometry. The viability of B16F10 was measured by the 3-（ 4,5- dimethyl- 2- thiazolyl）- 2,5- diphenyl- 2- H- tetrazolium bromide（ MTT）. Results Compared with the control group,the tumor volume and weight of Bre group,Cis group and Bre ＋ Cis group obviously reduced（ P〈0. 05,or P〈0. 01）,and the melanoma cell proliferation and angiogenesis rate were significantly inhibited（ P〈0. 05 or P〈0. 01）. The melanoma cell apoptosis rate were（ 13. 78 ± 1. 66） %,（ 12. 90 ± 1. 10） %,（ 18. 74 ± 1. 22） % for Bre,Cis and Bre ＋ Cis groups,respectively and compared with the control group（ 2. 02 ± 0. 32） %,the difference is statistically significant（ P〈0. 01）. Conclusion Breviscapine and cisplatin can inhibite the melanoma cell grow through their influences on thecell cycle and the formation of tumor blood vessels.