萤火虫荧光素酶标记结直肠癌模型的建立  

Construction of Colorectal Cancer Model with the Stable Expression of Firefly Luciferase

作　　者：杨俊寒 段海潇 张紫怡 王润杨 刘滨磊 胡翰 汪洋 YANG Junhan;DUAN Haixiao;ZHANG Ziyi;WANG Runyang;LIU Binlei;HU Han;WANG Yang(School of Biological Engineering and Food Science,Hubei Univ.,of Tech.Wuhan 430068,China)

机构地区：[1]湖北工业大学生物工程与食品学院,湖北武汉430068

出　　处：《湖北工业大学学报》2021年第2期46-49,共4页Journal of Hubei University of Technology

摘　　要：针对小鼠结直肠癌细胞(CT26)构建能够稳定表达萤火虫荧光素酶Firefly luciferase(Fluc)以及具有嘌呤霉素抗性的CT26-Fluc细胞株,并建立其荷瘤鼠模型。采用同源重组技术将Firefly luciferase基因片段整合至转座子系统中得到质粒,将所得质粒转染进入CT26原始细胞株,使其在该细胞内表达。后通过细胞敏感性实验获得CT26最低致死浓度,筛选获得稳定表达荧光素酶的CT26-Fluc单克隆细胞系。流式细胞仪检测CT26-Fluc单克隆的纯度。同时利用显微观察等方法证实细胞增殖和细胞形态与CT26原始细胞系不存在显著差异。将构建成功的CT26-Fluc植入BALB/c小鼠右后侧背部皮下,通过小动物活体成像仪验证该细胞系能稳定表达荧光素酶,且能够用于小鼠结直肠癌成瘤模型。本实验成功构建了稳定表达Firefly luciferase的CT26-Fluc细胞系,为小鼠结直肠癌模型及其病灶转移机制的研究建立了技术基础。The tumor bearing mouse model was established for mouse colon cancer cells(CT26),which could stably express Firefly luciferase(Fluc)and CT26 Fluc cell lines with puromycin resistance.The Firefly luciferase gene fragments were integrated into the transposon subsystem by homologous recombination technique to obtain the plasmid,and the resulting plasmid was transfected into the CT26 cell line to make it express in the cell.CT26 Fluc monoclonal cell lines with stable luciferase expression were obtained by purinomycin screening.The purity of CT26-Fluc clones was detected by flow cytometry,and the cell proliferation and morphology were not different from the CT26 cell lines.The constructed CT26 Fluc cell line was implanted BALB/c mouse subcutaneous,and it was determined by in vivo fluorescence of animals that the cell line could stably express luciferase and be used in mouse colorectal cancer tumor model.This experiment successfully constructed CT26 Fluc cell lines with stable expression of Firefly luciferase.It contributes to the study of tumorigenesis and metastasis mechanism of mouse colorectal cancer.

关 键 词：小鼠结直肠癌细胞 萤火虫荧光素酶 荧光成像 

分 类 号：R153[医药卫生—营养与食品卫生学]