纳米雄黄抗B16小鼠恶性黑色素瘤和抗新生血管形成作用的研究  

Study on the anti-tumor and anti-angiogenesis actions of realgar nanoparticles on B16 malignant melanoma in mice

作　　者：席晓霞[1] 范临兰[1] 席栋宾[2] 魏虎来[1] 田永刚[1] 张景科[1] 程杰[1] 

机构地区：[1]兰州大学基础医学院医学实验中心甘肃省新药临床前研究重点实验室,甘肃兰州730000 [2]酒泉市人民医院,甘肃酒泉735000

出　　处：《中国兽医科学》2015年第1期97-103,共7页Chinese Veterinary Science

基　　金：甘肃省中医药科学技术研究项目(GZK-2010-17);甘肃省自然科学研究基金计划项目(1208RJZA183)

摘　　要：为了用活体成像技术对纳米雄黄体内外对小鼠恶性黑色素瘤细胞(B16)增殖的抑制作用和机制进行研究,以萤火虫荧光素酶(luciferase,Luc)基因标记小鼠B16黑色素瘤(B16-Luc)细胞,用生物发光法(bioluminescent method,BLM)检测细胞增殖活性;以细胞形态学及AnnexinⅤ+PI双标记法观察细胞凋亡情况。采用B16-Luc细胞接种雄性BALB/c小鼠腋窝制作黑色素瘤模型,用纳米雄黄[4mg/(kg·d)和8mg/(kg·d)]灌胃治疗20d,以小动物活体成像系统动态观察肿瘤细胞的生长情况。治疗末期处死动物,取出肿瘤组织,固定、制片、HE染色,光镜下观察组织形态变化。结果显示,体外和体内纳米雄黄呈时间和剂量依赖性地抑制小鼠B16-Luc黑色素瘤的生长(P<0.01);肿瘤组织制片观察时,经纳米雄黄治疗后肿瘤组织内微小血管显著减少(P<0.01),肿瘤组织内部呈显著的坏死改变。结论:经用活体成像技术研究,纳米雄黄体内外能抑制小鼠B16-Luc黑色素瘤细胞的增殖并诱导其凋亡,通过抑制肿瘤组织内新生血管的形成而发挥抗小鼠恶性黑色素移植瘤作用;与传统技术相比,活体成像技术的灵敏度更高,在监测肿瘤细胞的生长方面优势更大。In order to study the anti-proliferating action of realgar nanoparticles(nano-realgar)on malignant melanoma cells in mice(B16-Luc)in vitro and in vivo using the optical in vivo imaging,the firefly luciferase gene was transferred into murine B16 melanoma cells(B16-Luc cells)with a lentiviral vector.Bioluminescence(BLM)assay was used to detect the cellular proliferation.The morphological observation and AnnexinⅤ and propidium iodide(AnnexinⅤ+PI)double-labeling were employed to assess the cell apoptosis in vitro.The B16-Luc cells were orthotopically implanted into male BALB/c mice to establish the orthotopic transplanted model of mouse malignant melanoma.The tumor-bearing mice were treated with 4mg/(kg·d)and 8mg/(kg·d)nano-realgar by gavage once a day for 20 days,respectively.The optical in vivoimaging system was used to continuously and dynamically observe the melanoma cell growth.At the end of the treatment,animals were sacrificed and the tumor tissue was removed.The tumor tissue sections were fixed,stained with HE staining,and observed under light microscope.In result,BLM assay showed that nano-realgar inhibited the growth of murine melanoma in vivo at a time-and dose-dependent manner(P<0.01).The tumor tissue section examination showed that the numbers of microvessels in B16-Luc tumor tissue significantly reduced and the center of the tumor tissue showed significant necrosis after realgar nanoparticle treatment.In conclusion,the realgar nanoparticles significantly inhibit the proliferation of murine B16-Luc malignant melanoma cells and induces to apoptosis in vitro and in vivo,and expresses the anti-melanoma action via restraining angiogenesis in tumor tissue.Compared with the traditional technique,the optical in vivo imaging is higher in sensitivity with greater advantage in monitoring of the growth of tumor cells.

关 键 词：纳米雄黄 小鼠恶性黑色素瘤 活体成像技术 

分 类 号：S857.4[农业科学—临床兽医学]