应用活体成像技术对三氧化二砷抗小鼠4T1乳腺癌作用的研究  

Study on anti-tumor effect of arsenic trioxide on orthotopic breast cancer in mice by the optical in vivo imaging technology

作　　者：范临兰[1] 席晓霞[1] 魏虎来[1] 张强弩[2] 高飞云[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]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 [3]兰州大学基础医学院病理学研究所 [4],甘肃兰州730000

出　　处：《中国兽医科学》2013年第2期170-176,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);甘肃省自然科学基金项目(096RJZA034)

摘　　要：采用小动物活体光学成像技术研究三氧化二砷(As2O3)的抗小鼠乳腺癌作用,并比较小动物活体成像技术与常规技术的优劣性。以荧光素酶(luciferase,Luc)基因标记小鼠4T1乳腺癌(4T1-Luc)细胞,MTT比色法和生物发光法(BLM)检测细胞增殖活性,细胞形态学及AnnexinⅤ/PI双标记法观察细胞凋亡;4T1-Luc细胞接种小鼠乳腺脂肪垫制作原位乳腺癌模型,腹腔注射5、10mg/(kg.d)As2O3治疗20d,小动物活体成像系统动态观察肿瘤生长。肿瘤称重,HE和CD34免疫组化染色观测肿瘤细胞分裂、坏死和新生微血管。结果,BLM检测结果与MTT法高度相符,1、2、4、8、16μmol/L As2O3显著抑制4T1-Luc细胞增殖,并呈现典型的凋亡改变;As2O3在体内呈时间和剂量依赖性地抑制模型小鼠4T1-Luc乳腺癌的生长,活体成像法优于肿瘤重量法。肿瘤组织内核分裂细胞和微小血管减少,中心呈坏死改变。结果表明,As2O3体外诱导小鼠乳腺癌4T1细胞凋亡,通过减低肿瘤新生血管形成、促使其坏死在体内发挥抗乳腺癌作用;与传统技术相比,小动物活体成像技术具有非损伤、实时动态监察和高灵敏度的特点。To study the proliferation-inhibiting action of arsenic trioxide（As2O3） on murine breast cancer 4T1 cells in vivo and in vitro with the optical in vivo imaging,and the advantages of the optical in vivo imaging technology were compared with the conventional animal experimental methods.The firefly luciferase gene-transferred 4T1 breast cancer cells（4T1-Luc cells） were used as target cells.The cellular cell proliferation was detected with both MTT colorimetric assay and bioluminescence（BLM） assay,and the morphological observation and AnnexinⅤ and propidium iodide（AnnexinⅤ/PI） double-labeling were employed to assess the cell apoptosis in vitro.The 4T1-Luc cells were implanted orthotopically into the mammary fat pad of female BALB/c mice to establish the orthotopic breast cancer model,the tumor-bearing mice were treated with 5 mg/（kg·d） and 10 mg/（kg·d） As2O3 by introperitoneal injection once a day for 20 days,respectively,the optical in vivo imaging system was used to continuously and dynamically monitor the tumor growth.At the end of the treatment,the animals were killed,and the tumor tissue was removed and weighed.The tumor tissue sections were prepared with HE staining and CD34 immunocytochemistry staining to examine karyokinesis,necrosis and angiogenesis in tumors.In result,BLM assay,highly consisted with MTT assay,showed that 1,2,4,8 and 16μmol/L As2O3significantly inhibited the proliferation of 4T1-Luc cells,and the morphological observation and AnnexinⅤ/PI double-staining displayed the typical apoptotic characteristics in As2O3-treated cells.As2O3 significantly inhibited the growth of murine 4T1 orthotopic breast cancer in vivo at a time-and dose-dependent manner,and the tumor-growth measurement by optical in vivo imaging was better than by tumor weighing.After As2O3 administration,the pathological and immunocytochemical measurement showed that the numbers of mitotic cells and microvessels in tumor tissue markedly decreased,and at the center of tumor tissue the extensive nec

关 键 词：三氧化二砷 原位移植乳腺癌 活体成像技术 血管生成 细胞凋亡 

分 类 号：R73[医药卫生—肿瘤]