慢病毒介导荧光素酶表达的人鼻咽癌细胞小鼠模型的建立

Establishment of a Nude Mouse Model of Human Nasopharyngeal Carcinoma with SUNE1 Cells Expressing Luciferase Mediated by Lentivirus

目的:建立荧光素酶标记的人鼻咽癌细胞裸鼠模型,活体成像系统监测肿瘤的生长并与肿瘤的体积进行对比。方法:构建表达荧光素酶基因2(luc2)的慢病毒载体,与辅助质粒共转染293T细胞以制备慢病毒,感染人鼻咽癌SUNE1细胞后经嘌呤霉素筛选获得表达luc2的细胞株。活体成像设备体外检测不同数量细胞的发光强度,最后以5×106个细胞皮下接种BALB/c nu/nu裸鼠,活体成像系统动态记录接种后肿瘤的信号并与肿瘤的体积对比。结果:成功构建慢病毒表达质粒pLenti-luc2并包装出慢病毒颗粒,病毒感染后嘌呤霉素筛选6天得到鼻咽癌细胞株SUNE1-luc2。细胞株传代后有稳定的发光强度,且经活体检测的每秒光子数与细胞数成正相关(R2=0.96);活体成像观察发现裸鼠接种第2天接种部位的发光强度就达到3.2×108,而且成瘤过程中发光强度的变化与肿瘤大小一致。结论:成功构建适用于活体成像的人鼻咽癌SUNE1细胞的裸鼠成瘤模型,该模型从细胞接种开始即可有效动态监测鼻咽癌皮下瘤的生长及转移,从而为鼻咽癌的成瘤机制及药物干预研究提供一个新的手段。

Objective： To establish a nude mouse model with human nasopharyngeal carcinoma cells labeled with luciferase, and to monitor tumor growth using in vivo imaging system and compare with tumor volume. Methods： Recombinant lentiviral vector express- ing luciferase 2 gene was constructed. Lentivirus was produced by co-transfecting lentiviral plasmid and package plasmids into 293T cells. After lentivirus infected human nasopharyngeal carcinoma SUNE1 cell, SUNEI-luc2 cell line was obtained by screening with puromycin. Relationship between the luminescence intensity and the cell number of SUNEI-luc2 was identified by using in vivo imaging system. 5 ~ 106 cells of SUNE 1-1uc2 were subcutaneously inoculated into BALB/c nu/nu nude mice, the photons per second was dynam- ically detected in vivo with imaging system, and their tumor volume were compared. Results： Lentiviral expression plasmid pLenti-luc2 was constructed, and lentivirus particles were packed. Screened with puromycin for 6 days, cell line SUNEI-luc2 was obtained. The luminescence intensity of SUNEI-luc2 cells was stable after passage, and the luminescence intensity was positive correlated with cell number （R2=0.96）. Two days after subcutaneous injection, tumor signals reached at 5~ 106 cells by imaging system for SUNEI-luc2, and 3.2 x 108 for the luminous intensity, which was consistent with tumor volume during the growth of tumor. Conclusion： A nude mouse model of nasopharyngeal carcinoma that could be used in vivo imaging was successfully established, and tumor growth and metastasis could be dynamically monitored since cells inoculation for 2 day. The model provided a new mean for studying nasopharyngeal tumor growth and drug intervention.