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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 a lung cancer model of green fluorescent protein (GFP) transgenic mice and observe the expression of GFP in lung cancer cells after the induction of cancer. Methods Sixty SPF BALB / c green fluorescent protein transgenic mice (4-6 weeks old) were randomly divided into experimental (30 mg / kg dimethylnitrosamine group) and control (saline group) Only), male and female, using gavage exposure, exposure capacity of 3 ml / kg, twice a week for 8 weeks. At the 15th and the 22nd week, the expression of GFP and the expression of CK7 and CK14 in lung tissue were observed. Twenty-four SPF-grade BALB / c-nu / nu nude mice aged 4 to 6 weeks were transplanted subcutaneously into lung tissue. Two weeks later, the survival of transplanted tumor and the expression of GFP were observed. Results The method could induce the establishment of Balb / c mouse lung cancer model. The incidence of lung cancer was 100% (16/16) after 22 weeks. No other organ metastasis was found. CK14 is highly expressed in tumor tissue, while CK7 is negative in tumor tissue. Subcutaneous xenografts and frozen sections have strong GFP expression. Conclusion Long-term low-dose diethylnitrosamine exposure can induce lung cancer formation in Balb / c green fluorescent protein transgenic mice, and maintain the GFP expression in cancer cells.