Glypican-3 Targeting of Liver Cancer Cells Using Multifunctional Nanoparticles
Glypican-3 Targeting of Liver Cancer Cells Using Multifunctional Nanoparticles
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DOI：
10.2310/7290.2010.00048
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发表时间：
2011-01-01
影响因子：
2.8
通讯作者：
Zhang, Miqin
中科院分区：
文献类型：
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作者：
Park, James O.;Stephen, Zachary;Zhang, Miqin
原发性肝癌（肝细胞癌[HCC]）是最常见和最致命的恶性肿瘤之一，影像学对于准确检测、分期和治疗原发性肝癌至关重要。我们开发了一种新的多功能纳米颗粒（NP），专门针对glypican-3 (GPC3)， GPC3是一种在大多数hcc中过度表达的促进细胞生长的蛋白多糖。定量实时聚合酶链反应证实了GPC3在Hep G2（高）和HLF（可忽略）两种人HCC细胞中的差异表达。这些细胞用生物素偶联的GPC3单克隆抗体（α GPC3）处理，随后用超顺磁性氧化铁NPs偶联链霉亲和素和Alexa Fluor 647靶向。流式细胞术表明，只有表达gpc3的Hep G2细胞被这种α GPC3-NP偶联物特异性靶向（比非靶向NP平均荧光增加4倍），磁共振成像（MRI）实验也显示了类似的结果（3倍R-2弛度）。共聚焦荧光显微镜将α - GPC3 NPs定位在表达GPC3的Hep G2细胞的细胞表面。进一步的表征表明该结构带负电，单分散，50 nm NP，非常适合肿瘤靶向。这种gpc3特异性NP系统具有双模成像能力，可以增强MRI预处理，通过近红外荧光精确显示术中HCC，并有可能作为肿瘤靶向治疗的载体，改善患者预后。
Imaging is essential in accurately detecting, staging, and treating primary liver cancer (hepatocellular carcinoma [HCC]), one of the most prevalent and lethal malignancies. We developed a novel multifunctional nanoparticle (NP) specifically targeting glypican-3 (GPC3), a proteoglycan implicated in promotion of cell growth that is overexpressed in most HCCs. Quantitative real-time polymerase chain reaction was performed to confirm the differential GPC3 expression in two human HCC cells, Hep G2 (high) and HLF (negligible). These cells were treated with biotin-conjugated GPC3 monoclonal antibody (alpha GPC3) and subsequently targeted using superparamagnetic iron oxide NPs conjugated to streptavidin and Alexa Fluor 647. Flow cytometry demonstrated that only GPC3-expressing Hep G2 cells were specifically targeted using this alpha GPC3-NP conjugate (fourfold mean fluorescence over nontargeted NP), and magnetic resonance imaging (MRI) experiments showed similar findings (threefold R-2 relaxivity). Confocal fluorescence microscopy localized the alpha GPC3 NPs only to the cell surface of GPC3-expressing Hep G2 cells. Further characterization of this construct demonstrated a negatively charged, monodisperse, 50 nm NP, ideally suited for tumor targeting. This GPC3-specific NP system, with dual-modality imaging capability, may enhance pretreatment MRI, enable refined intraoperative HCC visualization by near-infrared fluorescence, and be potentially used as a carrier for delivery of tumor-targeted therapies, improving patient outcomes.