Establishment of enhanced green fluorescent protein+luciferase+LS174T cells and mouse multiple-site tumor models
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.
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