智能型金纳米棒荧光探针的制备及其肿瘤细胞成像研究
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摘要
开发能特异和灵敏检测基质金属蛋白酶-2(MMP-2)的智能型荧光分子探针,对于癌症的早期精准诊断与治疗至关重要。本文基于MMP-2特异性识别的多肽底物,利用一价铜离子催化的"点击"化学反应将荧光素(FITC)和多肽底物GPLGVRGY相偶联,与SH-PEG-COOH修饰的金纳米棒在EDC和Sulfo-NHS的作用下通过酰胺化反应,制备得到一种新的MMP-2特异性响应的荧光共振能量转移(FRET)金纳米棒探针GNR@FITC。体外重组酶检测实验、MTT实验及细胞共聚焦显微成像结果表明,该探针对MMP-2过表达小鼠乳腺癌细胞4T1表现出较好的特异性和检测灵敏度,具有进一步开发应用于乳腺癌早期诊断的极大潜力。
Developing a smart fluorescent probe which can specifically and sensitively detect matrix metalloproteinases-2( MMP-2) is vital for the early accurate diagnosis and treatment of cancers. In this study,fluorescein isothiocyanate( FITC) was firstly conjugated to the peptide substrate GPLGVRGY by using copper( I)-catalyzed"click"reaction,and subsequently reacted with gold nanorods decorated with SH-PEG-COOH under the catalysis of EDC and Sulfo-NHS through amidation reaction to afford a novel fluorescence resonance energy transfer( FRET)-based probe GNR@ FITC for specific detection of MMP-2. In vitro enzyme detection experiments,MTT assays and fluorescence imaging results showed that the probe shows good specificity and high sensitivity for the detection of MMP-2-overexpressed 4 T1 cells,which may have great potential for clinical application in early diagnosis of breast cancer.
Developing a smart fluorescent probe which can specifically and sensitively detect matrix metalloproteinases-2( MMP-2) is vital for the early accurate diagnosis and treatment of cancers. In this study,fluorescein isothiocyanate( FITC) was firstly conjugated to the peptide substrate GPLGVRGY by using copper( I)-catalyzed"click"reaction,and subsequently reacted with gold nanorods decorated with SH-PEG-COOH under the catalysis of EDC and Sulfo-NHS through amidation reaction to afford a novel fluorescence resonance energy transfer( FRET)-based probe GNR@ FITC for specific detection of MMP-2. In vitro enzyme detection experiments,MTT assays and fluorescence imaging results showed that the probe shows good specificity and high sensitivity for the detection of MMP-2-overexpressed 4 T1 cells,which may have great potential for clinical application in early diagnosis of breast cancer.
引文
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