摘要
目的建立绿色荧光蛋白(green fluorescent protein,GFP)转基因小鼠肺癌模型,观察诱癌后肺组织细胞GFP的表达情况。方法将60只4～6周龄SPF级BALB/c绿色荧光蛋白转基因小鼠随机分为实验(30 mg/kg二甲基亚硝胺)组(50只)和对照(生理盐水)组(10只),雌雄各半,采用灌胃方式进行染毒,染毒容量为3 ml/kg,每周2次,连续8周。分别于第15、22周,观察小鼠肺组织GFP的表达情况和CK7、CK14表达情况。取肺肿物制成组织碎块移植于2只4～6周龄SPF级BALB/c-nu/nu裸鼠皮下,2周后观察移植瘤的存活与GFP的表达情况。结果本方法可以诱导建立Balb/c小鼠肺癌模型,22周以后肺癌的发生率为100%(16/16),未见其他脏器转移。CK14在肿瘤组织中高表达,而CK7在肿瘤组织中表达阴性。皮下移植瘤与冰冻切片均有较强的GFP表达。结论长期低剂量二乙基亚硝胺染毒可诱导Balb/c绿色荧光蛋白转基因小鼠肺癌形成,且癌细胞保持GFP表达。
Objective To establish the lung cancer model of green fluorescent protein （GFP） transgenic BALB/c mouse and observe the expression of green fluorescent protein in the lung tissue after successfully inducing cancer. Methods 4-6 weeks old male and female GFP transgenic mice（n=60） were randomly divided into an experimental group （n=50） and a control group（n=10）. The mice were treated with diethylnitrosamine through gavage, 30 mg/kg body weight in distilled water, twice a week for eight consecutive weeks and then the treatment was stopped, the mice were fed for another 12 weeks. The lung tissue was removed for frozen section to observe the GFP expression by fluorescence microscope, stained with hematoxylin and eosin for histopathological examinaion and immunohistochemical analysis of CK7 and CK14 at the 15th week and the 22rd week rspectively. In addition, the tissue fractions were transplanted into the subcutaneous of the nude mice for preparing the tumor-bearing mice. The biofluorescent signals and survival of xenograft were detected in sequential two weeks using in vivo fluorescent imaging system. Results The lung cancer model of fluorescent BALB/c mouse was successfully established by 0.3% DENA gavage. The inducing rate of tumor was 65% at the 22rd week after treatment with diethylnitrosamine. Tumors manifested positive expression of CK14 and negative expression of CKT,none of which combined with extrapulmonary metastasis. Frozen sections and subcutaneous tumors strongly expressed green fluorescent protein. Conclusion Long-term and low-dose diethylnitrosamine exposure can induce BABIJc mouse lung cancer. The cells of lung cancer can present stable high-level GFP expression in vivo.
出处
《环境与健康杂志》
CAS
CSCD
北大核心
2012年第8期686-689,F0003,共5页
Journal of Environment and Health
基金
广西自然科学基金(桂科自0991139)
广西科学研究与技术开发计划项目(桂科攻0993003C-1)
广西科学研究与技术开发计划项目(09-090-05)
关键词
绿色荧光蛋白
小鼠
肺癌
动物模型
活体成像系统
Green fluorescent protein （GFP）
Mice
Lung cancer
Animal model
Whole-body fluorescent imaging system
