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目的构建环状RGD肽偶联荧光素酶(luciferase)蛋白的荧光分子探针RGD-Luc,探讨RGD-Luc在体外与人肝癌细胞结合后分子显像对肝癌早期诊断的价值。方法⑴用化学方法偶联luciferase蛋白的氨基与环RGD短肽的巯基,并用Western Blotting鉴定;⑵将RGD-Luc、Luciferase分别与Hep G2细胞孵育2 h,同时设立未作处理的对照组,使用小动物活体成像系统分别观察靶向结合情况。结果⑴Western Blotting分析证明荧光素酶luciferase与c RGDf C肽成功偶联;⑵小动物活体成像系统检测示RGD-Luc荧光探针能够特异性的与Hep G2细胞结合,而Luciferase组及对照组未观察到荧光分布。结论利用化学偶联方法成功制备新型荧光探针RGD-Luc,其能靶向增强肝癌细胞的分子显像,为肝癌的早期显像提供了一定的实验依据。
OBJECTIVE: To construct RGD-Luc fluorescent probe for RGD peptide-coupled luciferase protein and to explore the value of molecular imaging of RGD-Luc binding to human hepatoma cells in vitro for the early diagnosis of liver cancer. Methods (1) The amino groups of luciferase protein and RGD short peptide were coupled by chemical methods and identified by Western Blotting. (2) RGD-Luc and Luciferase were respectively incubated with Hep G2 cells for 2 h. At the same time, untreated control group Live animal imaging system to observe the targeted binding. Results (1) Luciferase luciferase and c RGDf C peptide were successfully coupled by Western Blotting assay. (2) Live animal imaging system showed that RGD-Luc fluorescent probe could specifically bind to Hep G2 cells, while Luciferase group and control group were not observed To the fluorescence distribution. Conclusion The new fluorescent probe RGD-Luc was successfully prepared by chemical coupling method, which can enhance the molecular imaging of hepatoma cells and provide some experimental evidence for the early imaging of liver cancer.