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目的探讨七甲川花菁近红外荧光(near-infrared fluorescence,NIRF)染料IR-783在肝癌模型影像研究中的应用,分析该染料靶向肿瘤细胞的分子机制。方法将标记荧光素酶的人肝癌细胞Hep G2皮下接种裸鼠,测试肿瘤发生部位生物发光(bioluminescence,BIL)信号与NIRF信号的相关性。采用肾包膜下移植的方式建立人肝癌裸鼠移植(patient-derived xenograft,PDX)模型,观察IR-783对肿瘤边界的识别能力;苏木素-伊红染色(hematoxylin-eosin staining,H&E staining)确认肿瘤发生的部位,免疫组织化学(Immunohistochemistry,IHC)检测肝癌肿瘤组织中CEA、AFP、HIF1α和OATP3A1的表达;使用线粒体示踪剂(Mito Tracker)和溶酶体示踪剂(Lyso Tracker)确认NIRF染料在肝癌细胞中的结合部位;测试IR-783对正常肝细胞中混合培养的肝癌细胞鉴别能力。结果人肝癌裸鼠皮下移植瘤BIL与NIRF强度具有较好的相关性;NIRF染料IR-783能够清晰识别肾包膜移植肝癌肿瘤边缘;IHC染色显示肿瘤组织中CEA、AFP、HIF1α和OATP3A1均高表达;IR-783主要结合在肿瘤细胞的线粒体与溶酶体中;标记GFP的人肝癌细胞Hep G2能够被IR-783特异性识别。结论 IR-783是一种具有肿瘤靶向和成像特征的新型近红外荧光染料,其靶向性可能与肝癌组织中HIF1α和OATP3A1的高表达相关。
Objective To investigate the application of near-infrared fluorescence (NIRF) dye IR-783 in the research of hepatocellular carcinoma (HCC) model and analyze the molecular mechanism of this dye targeting tumor cells. Methods Human hepatoma Hep G2 cells labeled with luciferase were subcutaneously inoculated with nude mice to test the correlation between the bioluminescence (BIL) signal and NIRF signal. The model of patient-derived xenograft (PDX) was established by subrenal transplantation, and the ability of IR-783 to recognize tumor border was observed. H & E staining was confirmed The expression of CEA, AFP, HIF1α and OATP3A1 in hepatocellular carcinoma tissues was detected by immunohistochemistry (IHC). NIRF was confirmed by Mito Tracker and Lyso Tracker Dye binding sites in liver cancer cells; test IR-783 on normal liver cells in mixed culture of liver cancer cells ability to identify. Results The BIL and NIRF intensity of human hepatocellular carcinoma subcutaneously transplanted in nude mice had a good correlation. NIRF dye IR-783 could clearly identify the edge of renal carcinoma with renal capsule graft. The IHC staining showed that CEA, AFP, HIF1α and OATP3A1 were all high . IR-783 mainly binds to mitochondria and lysosomes in tumor cells. Hep G2, a GFP-tagged human hepatoma cell, can be specifically recognized by IR-783. Conclusion IR-783 is a novel near-infrared fluorescent dye with tumor targeting and imaging features. Its targeting ability may be related to the high expression of HIF1α and OATP3A1 in hepatocellular carcinoma.