ABCE1-GFP肺腺癌动物模型的建立及免疫组化检测

The establishment of lung adenocarcinoma animal model expressing ABCE1- GFP and immunohistochemical detection

目的:建立ABCE1-GFP(+)肺腺癌裸鼠动物模型,并进行免疫组化检测,为进一步探索ABCE1作为诊断治疗非小细胞肺癌靶点提供有用的工具。方法:应用前期实验中构建的稳定转染重组质粒p EGFPC1-ABCE1及空载体p EGFP-C1的LTEP-a-2肺腺癌细胞株,接种于裸鼠背部侧翼至腋窝皮下,建立ABCE1-GFP肺腺癌动物模型。进行成瘤情况观察,并予以倒置荧光显微镜系统呈像。取材后进行免疫组织化学检测。结果:稳转细胞系皮下接种裸鼠后,全部成瘤,过表达组肿瘤体积大于空载体及对照组(P<0.05)。过表达组肿瘤在荧光显微镜系统下可稳定表达绿色荧光,成功构建了ABCE1-GFP(+)肺腺癌皮下成瘤的裸鼠动物模型。免疫组织化学检测亦验证这一模型的确立。结论:稳定转染LTEP-a-2肺腺癌细胞可以用来成功建立皮下ABCE1-GFP(+)肺腺癌成瘤模型,是较为理想研究ABCE1在非小细胞肺癌中的基因功能的动物模型。

Objective： To establish ABCE1- GFP（ ＋） tumor animal model of nude mice and test it by immunohistochemistry,in order to provide an useful tool for further exploring ABCE1 gene as a non- small cell lung cancer diagnosis and treatment target. Methods： We inoculated subcutaneously lung adenocarcinoma cell line LTEP- a- 2stably expressing p EGFP- C1- ABCE1 and p EGFP- C1 into BALB / C- nude mice,establishing ABCE1- GFP（ ＋） tumor animal model,then observed tumor formation and took pictures by fluorescence microscope system. After that tumors were tested by immunohistochemical detection. Results： All the nude mice formed tumors subcutaneously,and the volume of overexpression group was demonstrated bigger than empty vector or control group（ P〈 0. 05）. Moreover,the tumors emitted green fluorescence in vivo. We succeed in establishing ABCE1- GFP（ ＋） tumor animal model of nude mice. Conclusion： Lung adenocarcinoma cell lines stably expressing plasmid p EGFP- C1- ABCE1 could establish ABCE1- GFP（ ＋） tumor animal model. Our research validated the model as an ideal tool for studying ABCE1 gene functions in non- small cell lung cancer.