采用活体成像技术监测肿瘤生长及转移模型的建立

A Novel Animal Model Established Using in Vivo Imaging to Monitor the Tumor Growth and Metastasis

目的采用活体成像技术监测稳定高表达荧光素酶报告基因的肿瘤细胞在小鼠体内生长及转移情况,为肿瘤治疗的药物研发提供新的有用工具。方法采用lipofectamine2000介导的基因转染方法,将pcDNA3·1/Luc载体转染小鼠高转移乳腺癌细胞株4T1、EMT-6及结肠癌细胞株CT26,经G418抗性筛选及有限稀释法获得可稳定高表达荧光素酶的单克隆细胞;MTT法测定各转染细胞对不同化疗药物的抗性,并采用活体成像的方法检测各转染细胞在小鼠体内的成瘤和转移。结果获得了可稳定高表达荧光素酶基因的单克隆细胞株,该单克隆细胞株具有与亲本细胞系相同的对化疗药物的敏感性;将单克隆细胞株植入小鼠皮下,可采用活体成像技术准确监测肿瘤细胞体内生长及转移。结论采用活体成像技术构建的肿瘤动物模型是拓展肿瘤体内生长、转移及治疗相关研究的理想模型。

Objective To monitor the in vivo growth and metastasis of tumors that were stably transfected with the report gene, luciferase, and to establish a novel tool for the development of new anti-tumor drugs. Methods Highly metastatic tumor cell lines, 4T1, CT26 and EMT-6, were transfected with plasmids pcDNA3.1/Luc using lipofectamine2000. Resistant clones were screened using G418. Stably transfected monoclones were obtained using limiting dilution analysis （LDA） and their resistance to anti-tumor drugs was tested using MTT assay. These stably transfected monoclones were subcutaneously transplanted into mice, and their tumorigenesis, metastasis were analyzed using in vivo imaging assessment. Results Stably transfected monoclones highly expressing Luc were obtained, which had the same sensitivity to anti-tumor drugs as their parental cell lines. After stably transfected monoclones being subcutaneous transplanted into the mice, the tumorigenesis and metastasis of monoclonal tumor cells were accurately monitored by in vivo imaging. Conclusions This novel animal model established using in vivo imaging may serve the further research of growth and metastasis of tumors, and development of anti-cancer drugs.