人肝癌原位移植瘤的活体动态观察

目的 建立GFP（绿色荧光蛋白）/Luciferase（荧光素酶,Luc）双标高效表达的人肝癌细胞系SMMC-7721-GFP/Luc,并建立该细胞的裸小鼠肝原位移植瘤动物模型,利用小动物活体成像系统观察裸小鼠肝原位移植瘤的生长及转移情况.方法 应用慢病毒转染的方法建立GFP/I.alC双标的SMMC-7721细胞系,接种于裸小鼠肝原位,采用活体成像技术监测该细胞在小鼠体内深部组织的表达情况.结果 成功建立了GFP/Luc双标表达率接近100%的SMMC-772l-GFP/Luc细胞系,进行裸小鼠肝原位移植后,应用活体成像系统可观察到裸小鼠肝原位早期的微小病灶和晚期移植瘤的生长情况.而且,随着肿瘤移植时间的延长,移植瘤体积的增大,活体成像检测到的发光面积逐渐增大,发光强度也逐渐增加.结论 活体荧光成像系统可直接观察肝原位移植瘤模型中肿瘤的生长及转移,有助于了解人肝癌的生物学行为及其特性,鉴于其敏感、直观,动态、可靠的优点,它将为肝癌的进一步研究提供极其有益的帮助.

Dynamic Observation of In vivo Bioluminescence Imaging of Orthotopic Implantation of Human Hepatocellular Carcinoma in Nude Mice

Objective To establish a human hepatocellular carcinoma cell line with stable and high- level GFP/Luciferase（Luc） expression, and observe the development of tumors in deep organisms of the nude mice by in vivo biofluorescence imaging. Methods The SMMC-7721-GFP/Luc cell line were established by lentivirus transfection, then were orthotopic implanted in the liver of the BALB/c nude mice, the behaviors of the tumors was detected by in vivo bioluminescetace imaging. Results The cell line SMMC-7721-GFP/Luc with stable and high-level GFP/Luc expression had been successfully established. After the cell line had been inoculated in the liver of the nude mice, the tiny tumor masses in early time and the growth of tumors in late time in the deep organisms were observed by in vivo biofluorescence imaging. Conclusion The use of cell line SMMC-7721-GFP/Luc could directly observe the tumor growth and metastasis in the orthotopic implantation animal model. It would help us to understand the biological behavior of human hepatocellular carcinoma and its characteristics. In view of its sensibility, intuitiveness, dynamics and reliability, it would do some good to further study of human hepatocellular carcinoma.