Biosens Bioelectron. 2014 Jul 15;57:171-8. doi: 10.1016/j.bios.2014.02.011. Epub 2014 Feb 19.
Molecular recognition of proteolytic activity in metastatic cancer cells using fluorogenic gold nanoprobes.
Hong Y1, Ku M2, Heo D3, Hwang S3, Lee E3, Park J4, Choi J5, Lee HJ6, Seo M7, Lee EJ6, Yook JI8, Haam S9, Huh YM10, Yoon DS1, Suh JS11, Yang J12.
Abstract
We describe the development of biomarker-sensitive nanoprobes based on nanoparticle surface energy transfer (NSET) effect that enabling recognition of the expression of membrane type-1 matrix metalloproteinase (MT1-MMP) anchored on invasive cancer cells and its proteolytic activity simultaneously. First of all, we confirmed invasiveness of cancer cell lines (HT1080 and MCF7) via migration and invasion assay. We also prepared gold nanoparticle (GNP) acts as a quencher for fluorescein isothiocyanate (FITC). This FITC is conjugated in end-terminal of activatable fluorogenic peptide (ActFP). The ActFP attach to surface of GNP (GNP-ActFP) for a targeting moiety and proteolytic activity ligand toward MT1-MMP. The GNP-ActFP can generate fluorescence signal when ActFP is cleaved by proteolytic activity after targeting toward MT1-MMP. In order to study specificity for MT1-MMP, GNP-ActFP is treated to HT1080 and MCF7 cells, and then, we determine the in vitro targeting potential and fluorogenic activity of GNP-ActFP for MT1-MMP via fluorescence multi-reader. We also confirmed fluorogenic activity of GNP-ActFP via confocal microscopic imaging, and finally, endocytosis of GNP-ActFP is observed via cellular transmission electron microscopic imaging.
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KEYWORDS:
Cancer metastasis; Fluorescence; Membrane type 1-matrix metalloproteinase (MT1-MMP); Nanoparticle surface energy transfer (NSET); Nanoprobe; Proteolysis