生物发光人肝癌裸小鼠原位癌模型的构建

Establishment of an nude mouse orthotopic tumor model of human hepatocellular carcinoma with luciferase expression

目的选用稳定表达荧光素酶报告基因的人源性肝癌细胞株(HCCLM3-Luc),构建人肝癌细胞株原位癌荷瘤裸鼠动物模型。方法活体成像仪定量分析不同细胞数量HCCLM3-Luc的荧光强度表达情况,并检测各细胞数量的荧光素酶活性,探索HCCLM3-Luc荧光素酶产生的光子量与肿瘤细胞数量之间的线性相关性。采用肝叶原位接种HCCLM3-Luc瘤块组织和尾静脉注射HCCLM3-Luc细胞悬液,分别构建人肝癌裸小鼠原位癌模型;比较两种方法构建人肝癌细胞株原位癌裸鼠模型的可行性,并优化模型构建中的关键因素。结果HCCLM3-Luc细胞数量的成正相关,(相关系数R^2=0.9989,直线方程式:Y=1155.8X+1×10^6),平均ROI值约为每个细胞140光子/s。采用肝叶原位接种组的原位瘤成瘤率[(82.5±7.2)%]明显高于尾静脉注射[(34.1±13.2)%]。结论本课题构建了生物发光人肝癌细胞株原位癌裸鼠模型,该模型可采用小动物成像仪实时监测肿瘤生长情况。

Objective A human hepatocellular carcinoma cell line(HCCLM3-Luc)stably expressing a luciferase reporter gene was used to establish an orthotopic mouse model.Methods An imaging system for living animals was used to quantitatively analyze the bioluminescence intensity of HCCLM3-Luc cells,which reflects the luciferase activity in cells.Then,the linear correlation between the photon quantity generated by HCCLM3-Luc luciferase and the number of HCCLM3-Luc cells was explored.The nude mouse model of orthotopic human hepatocellular carcinoma was constructed by in situ inoculation of HCCLM3-Luc tumor tissue in the hepatic lobe or injection with HCCLM3-Luc cell suspension through the tail vein.The two methods were evaluated and compared based on the tumor formation rate,the time of formation and the change of tumor shape.Results The bioluminescence intensity of the cells was positively correlated with the number of cells(correlation coefficient R^2=0.9989,linear equation:Y=1155.8X+1×10^6),and the average ROI value was approximately 140 photons/s per cell.Conclusions In this study,an orthotopic hepatocellular carcinoma-bearing mouse model using a bioluminescent human hepatocellular carcinoma cell line was established.The tumor growth was assessed by measuring the bioluminescence in real time using the IVIS Lumina XR Imaging System.