VEGF靶向分子探针的制备及对卵巢癌细胞SKOV3体外成像的初步研究

Preparation of VEGF Targeted Molecular Probes and Preliminary Study on in Vitro Imaging of Ovarian Cancer Cells SKOV3

目的:制备分子探针VEGF-USPIO和研究其在体外对卵巢癌细胞的靶向成像作用。方法:采用化学偶联法将VEGF抗体与USPIO连接,构建成具有免疫活性的靶向分子探针VEGF-USPIO。用CCK-8法检测该探针对卵巢癌细胞株SKOV3细胞活性的影响;普鲁士蓝染色法检测细胞磁性标记情况,并对经过标记的细胞进行体外磁共振成像,观察其对磁共振信号强度的影响。结果:成功合成了分子靶向探针VEGF-USPIO;当探针浓度在60μg/mL及以下时对细胞活性无影响(P>0.05);细胞普鲁士蓝染色结果显示标记了靶向探针VEGF-USPIO的细胞其胞膜及胞浆含铁颗粒沉积较多;细胞体外磁共振成像结果显示经靶向探针标记的细胞,T2WI信号强度较未标记探针的对照组细胞降低(P<0.05)。结论:所合成的VEGF-USPIO通过磁共振信号强度的变化实现MRI成像。

Objective: To prepare a specific targeting VEGF-USPIO molecular probe and explore its target imaging effect on ovarian cancer cells in vitro. Methods: The VEGF antibody was ligated with USPIO using a chemical coupling method to construct an immunologically active targeting molecule probe VEGF-USPIO. The effect of the probe on the activity of ovarian cancer cell line SKOV3 by CCK-8 assay. The Prussian blue staining method was used to detect the magnetic labeling of the cells, and the labelled cells were subjected to in vitro magnetic resonance imaging to observe their influence on the magnetic resonance signal intensity. Results: The molecular targeting probe VEGF-USPIO was successfully synthesized. The results of cytotoxicity test showed that it had no effect on cell viability when the probe concentration was 60 μg/mL or less, (P >0.05); Prussian blue staining showed that the cells labeled with VEGF-USPIO targeting probe had more deposition of iron-containing particles in the plasma membrane and cytoplasm. In vitro magnetic resonance imaging (MRI) showed that the T2WI signal intensity of the cells labeled with the targeted probe was lower than that of the control cells without the labeled probe (P<0.05). Conclusion: The synthesized molecularly-targeted probe VEGF-USPIO achieve specific MRI imaging through changes in signal intensity.