荧光素酶标记的BEL-7405细胞体内外成像及肿瘤动物模型建立

Bioluminescent Imaging of BEL-7405-luciferase Cells and Model Establishment of Tumor-animal

目的筛选表达荧光素酶(luciferase)基因的人单克隆肝癌细胞系,用活体成像技术(bioluminescentimaging)在细胞及整体动物水平检测其发光能力,为监测肿瘤生长和转移建立一种新的肿瘤动物模型。方法构建表达荧光素酶基因的真核表达载体并将其转入肝癌细胞(BEL-7405),用G418筛选稳定表达荧光素酶的单细胞克隆,在体外用活体成像技术评价其稳定发光能力。BALB/c nu/nu裸鼠皮下接种1×106个发光细胞使其成瘤,活体内观察肿瘤的生长及转移情况。结果在肝癌细胞中获得稳定表达荧光素酶的克隆,利用活体成像技术可检测到细胞克隆发光。将稳定表达荧光素酶的克隆皮下接种到裸鼠体内可成瘤,利用活体成像系统观察了肿瘤的生长过程,肿瘤发光随着观察时间的延长而增强,但是没有观察到肿瘤转移的现象。结论本研究筛选得到了稳定表达荧光素酶的肿瘤单克隆细胞系,结合活体成像技术,建立了一套新的能够用于活体内监测肿瘤生长的研究方法。

Objective To establish a new model for monitoring noninvasive growth and metastasis of tumor in animals. We screened a luciferase-expressing cell lines from the standard human liver tumor cell lines （ BEL-7405 ） and evaluated their bioluminescent ability in vitro and tumorigenesis ability in vivo with bioluminescent imaging technology. Methods The vector containing luciferase gene was constructed and transfected into liver tumor cell lines and selected with G418.The bioluminescent ability of stably luciferase-expressing cells was evaluated by BLI technology. 1 × 10^6 cells, constitutively expressing luciferase, were inoculated under skin of BALB/c nu/nu mice for assessment of tumour growth and metastasis. Result We have successfully got luciferase-expressing cell clones BEL-7405-luc ＋ in liver tumor cell lines, Bioluminescent imaging technology was used to measure their bioluminescent ability, subcutaneous inoculate BEL-7405-luc ＋ below dorsal blank of BALB/c nu/nu to observe growth of the tumor and metastasis ability. Conclusion We have got the clone of stably expressing luciferase and established a new set of animal model that can be used to monitor the tumor growth.