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

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)×10^(8)p/s/cm^(2)sr比(9.648±0.320)×10^(7)、0 p/s/cm^(2)sr,t=5.030、6.249,P<0.01];第14天肝转移肿瘤平均荧光强度为1.244×10^(7)p/s/cm^(2)sr。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。

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-Luciferase-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 system.Independent sample t-test was used for group comparison.Results Inoculation of EGFP+Luciferase+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)×10^(8)p/s/cm^(2)sr,vs.(9.648±0.320)×10^(7),0 p/s/cm^(2)sr,t=5.030,6.249,P<0.01].The average radiance of liver metastatic tumor groupon Day14 was 1.244×10^(7)p/s/cm^(2)sr.Conclusion Utilization of EGFP+Luciferase+LS174T in establishing mouse multiple-site tumor models enables efficient labeling and dynamic monitoring of tumor growth.Fluorescence intensity of tumors increased progressively with the duration of observation.