七甲川菁荧光染料在肝细胞癌移植模型活体成像中的应用

The Application of Heptamethine Cyanine Dye in Optical Imaging of Hepatocellular Carcinoma Transplantation Model

目的探索七甲川菁近红外(NIR)荧光染料MHI-148在肝癌移植模型中的代谢特征,明确其识别肝细胞癌(HCC)细胞的特异性。方法体外培养人HCC细胞系Hep3B,荧光显微镜下观察荧光染料MHI-148在肿瘤细胞中富集强度;裸小鼠皮下注射Hep3B细胞建立移植模型,2周后,荷瘤鼠腹腔注射MHI-148溶液,NIR荧光活体成像检测不同时间点肿瘤部位和裸小鼠脏器中荧光强度的变化;连续检测肿瘤部位荧光强度(tumor,T)和临近正常组织荧光强度(background,B),计算最大T/B值。结果 MHI-148染料可特异性集聚于体外培养的HCC细胞,识别绿色荧光蛋白(GFP)标记的Hep3B细胞;肝癌皮下荷瘤鼠注射MHI-148,24 h后肿瘤部位荧光强度可达8.35×109/cm2,而小鼠其他脏器较少有荧光聚集;T/B值逐渐增加,并在24 h达到峰值。结论七甲川菁染料MHI-148可特异性识别人HCC细胞系Hep3B,通过NIR荧光成像或可检测到肝癌的发生。

Objective To explore the metabolic characteristics of heptamethine cyanine near-infrared（NIR） fluorescence dye MHI-148 in hepatocellular carcinoma（HCC） xenograft model. Methods Human HCC cell Hep3 B was co-cultured with NIR dye MHI-148 solution. The specific accumulation of MHI-148 on tumor cells was observed under fluorescence microscope. Hep3 B cells were implanted subcutaneously into nude mice to establish a transplant model. Two weeks later, MHI-148 dye was injected intraperitoneally into tumor bearing nude mice and detected by NIR fluorescence optical imaging. The fluorescence intensity both on the tumor site and the organs of the nude mice was measured at different time point, the tumor fluorescence intensity（tumor, T） and the adjacent normal tissue（background, B） was continuously detected and the maximum T/B value was calculated. Results MHI-148 dyes could be specifically accumulated in HCC cells, and Hep3 B cells labeled with green fluoresence protein（GFP） were identified.When MHI-148 was injected into nude mice bearing subcutaneous HCC tumor, the fluorescence intensity per unit area（cm2） of the tumor site reached 8.35 × 109 after 24 hours, while other organs have less fluorescence aggregation. The ratio of T/B at the site of the tumor gradually increased and reached peak value at 24 hours. Conclusion The hepatocellular carcinoma cells can be specifically recognized by heptamethine cyanine dye MHI-148. The site of subcutaneous HCC tumor can be identified through NIR fluoresence optical imaging.