海肾荧光素酶基因标记肝癌细胞生物发光成像在小鼠肝癌活体示踪中应用  

Application of bioluminescence imaging of renilla luciferase labelling liver cancer cells in mice in vivo liver cancer tracking

作　　者：张丽娜[1] 李征然[1] 黄明声[1] 王劲[1] 郭若汨[1] 唐文杰[1] Zhang Lina;Li Zhengran;Huang Mingsheng;Wang Jin;Guo Ruomi;Tang Wenjie(Department of Radiology, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China)

机构地区：[1]中山大学附属第三医院放射科,广州510630

出　　处：《中华肝脏外科手术学电子杂志》2017年第6期504-508,共5页Chinese Journal of Hepatic Surgery(Electronic Edition)

基　　金：国家自然科学基金(81271621;81172193);广东省科技计划项目(2009B030801026)

摘　　要：目的探讨海肾荧光素酶(Rluc)标记肝癌细胞生物发光成像在小鼠肝癌活体示踪中应用价值。方法构建Rluc基因标记的Rluc-GFP-Hepa1-6肝癌细胞,利用流式细胞仪分选GFP表达阳性的稳定细胞;Rluc-GFP-Hepa1-6细胞行细胞生物发光成像,分析生物发光信号强度与细胞数量间相关性;同时将Rluc-GFP-Hepa1-6细胞接种于裸鼠皮下,建立移植瘤模型,行活体生物发光成像,定量监测肿瘤细胞在体内的生长过程。发光信号强度与细胞数量及肿瘤体积间关系采用线性回归分析。结果成功构建Rluc-GFP-Hepa1-6细胞株,经流式细胞仪分选后获得GFP阳性率高达95%的Rluc-GFP-Hepa1-6细胞。细胞生物发光成像结果显示,Rluc-GFP-Hepa1-6细胞发光信号强度与细胞数量间有相关性(R^2=0.999)。活体生物发光成像显示,皮下肿瘤的发光信号强度与肿瘤体积存在正相关性(R^2=0.887)。结论应用Rluc生物发光成像可成功监测小鼠肝癌皮下肿瘤模型的演进过程,不仅为研究肝癌体内生长、转移、治疗提供了理想的模型,而且为进一步研究肝癌的治疗效果提供了良好、无创的定量示踪手段。Objective To investigate the application value of bioluminescence imaging of ranillaluciferase(Rluc)labelling liver cancer cells in mice in vivo liver cancer tracking.Methods Rluc-GFPHepa1-6liver cancer cells were constructed.The stable cells with positive expression of GFP were purified byusing flow cytometry.Rluc-GFP-Hepa1-6cells were used for cell bioluminescence imaging.The correlationbetween the bioluminescence signal intensity and cell quantity was analyzed.The mice model with graftedtumors was constructed by subcutaneous inoculation of Rluc-GFP-Hepa1-6cells,and in vivo bioluminescentimaging was applied.The growth of tumor cells in the mice was quantitatively monitored.The correlationbetween the bioluminescence signal intensity,cell quantity and tumor size was analyzed using linearregression analysis.Results The Rluc-GFP-Hepa1-6cell lines were successfully constructed.The GFPpositive rate of Rluc-GFP-Hepa1-6cells purified by flow cytometry was95%.Cell bioluminescence imagingrevealed that the bioluminescence signal intensity of Rluc-GFP-Hepa1-6cells was significantly correlatedwith cell quantity(R2=0.999).In vivo bioluminescence imaging revealed that the bioluminescence signalintensity of subcutaneous tumors was significantly correlated with tumor size(R2=0.887).Conclusions Rluc bioluminescence imaging can successfully monitor the evolution of subcutaneous malignant tumors inmice model with liver cancer.It not only serves as an ideal model for studying the growth,metastasis andtreatment of liver cancer,but also provides an excellent,minimally invasive quantitative tracking approachfor evaluating the therapeutic efficacy of liver cancer.

关 键 词：癌 肝细胞 荧光抗体技术 生物发光成像 海肾荧光素酶 

分 类 号：R735.7[医药卫生—肿瘤]