双荧光标签LS174T细胞及小鼠多部位肿瘤模型的构建
Establishment of enhanced green fluorescent protein+luciferase+LS174T cells and mouse multiple-site tumor models
李屿堃 1吉登波 1顾晋2
作者信息
- 1. 北京大学肿瘤医院胃肠肿瘤中心三病区,北京 100142
- 2. 北京大学肿瘤医院胃肠肿瘤中心三病区,北京 100142;北京大学首钢医院胃肠外科,北京 100142
- 折叠
摘要
目的 构建表达增强型绿色荧光蛋白(EGFP)及荧光素酶的LS174T细胞,并建立多部位肿瘤小鼠模型,实现肿瘤细胞体内标记与动态监测.方法 使用质粒转染LS174T,经抗生素筛选及流式分选后获得EGFP+Luciferase+LS174T稳转系,接种于NOG免疫缺陷小鼠.皮下肿瘤组(5只)、腋下肿瘤组(3只)、肝转移肿瘤组(3只),使用活体成像监测肿瘤生长.组间比较采用独立样本t检验.结果 EGFP+Luciferase+LS174T在体外检测到EGFP表达,接种后5～7 d稳定成瘤.第9天腋下肿瘤组平均荧光强度(avg radiance)高于皮下肿瘤组及肝转移肿瘤组[(5.505±1.510)×108 p/s/cm2sr 比(9.648±0.320)×107、0 p/s/cm2sr,t=5.030、6.249,P＜0.01];第 14 天肝转移肿瘤平均荧光强度为1.244 ×107p/s/cm2sr.结论 使用EGFP+Luciferase+LS174T,建立多部位小鼠肿瘤模型均能实现体内肿瘤有效标记与示踪,肿瘤荧光强度随着观察时间的延长而增加.
Abstract
Objective To establish a colon cancer cell line LS174T stably expressing enhanced green fluorescent protein and luciferase,and multiple-site tumor models for in vivo labeling and dynamic monitoring of tumor growth.Methods LS174T was transfected with a plasmid(pLenti-CMV-EGFP-Lucif-erase-Hygro).EGFP+Luciferase+LS174T was selected by using hygromycin and flow cytometry.A total of 5 NOG mice were used to construct subcutaneous tumor models,three for axillary tumor models,and three for liver metastatic tumor models.Tumor growth was monitored by in vivo fluorescence imaging sys-tem.Independent sample t-test was used for group comparison.Results Inoculation of EGFP+Lucifer-ase+LS174T in NOG mice resulted in formation of tumorswithin 7 days.The average radiance of axillary tumor group on Day9 was higher than subcutaneous tumor group and liver metastatic tumor group[(5.505±1.510)× 108 p/s/cm2sr,vs.(9.648±0.320)× 107,0 p/s/cm2sr,t=5.030,6.249,P＜0.01].The average radiance of liver metastatic tumor groupon Day 14 was 1.244 × 107 p/s/cm2sr.Con-clusion Utilization of EGFP+Luciferase+LS174T in establishing mouse multiple-site tumor models ena-bles efficient labeling and dynamic monitoring of tumor growth.Fluorescence intensity of tumors increased progressively with the duration of observation.
关键词
结肠癌/荧光素酶/活体成像/肿瘤模型Key words
Colon cancer/Luciferase/In vivo imaging/Tumor models引用本文复制引用
出版年
2024
NETL
NSTL
万方数据