Profiling of multiple matrix metalloproteinases activities in the progression of osteosarcoma by peptide microarray-based fluorescence assay on polymer brush coated zinc oxide nanorod substrate

Abstract

Osteosarcoma (OS) is a primary malignant bone tumor with high rate of recurrence and lung metastasis. Matrix metalloproteinases (MMPs) are demonstrated as important biomarkers of tumor invasion and metastasis. Herein, a peptide microarray-based fluorescence assay is proposed to profile multiple MMPs (MMP-1, -2, -3, -7, -9 and -13) activities in the progression of OS by using the mouse-bearing xenograft U-2OS and Saos-2 human OSs. The peptide microarray containing different biotinylated peptide substrate spots is fabricated on the poly(glycidyl methacrylate-co-2-hydroxyethyl methacrylate) brush coated zinc oxide nanorod ([email protected](GMA-HEMA) decorated glass slides, and labeled by cyanine 3 (Cy3) modified avidin (Avidin-Cy3) to produce fluorescence signal. Special cleavage of peptide substrate by MMP resulted the decrease of fluorescence signal. The MMPs activities are positively correlated with the change of fluorescence intensity. This method has excellent selectivity and sensitivity, which enables to detect the activities of cellular secreted MMP-1, -2, -3, -7, -9, and -13 with limit of detection downs to 10 pmol L−1, 30 pmol L−1, 113 pmol L−1, 13 pmol L−1, 93 pmol L−1 and 12 pmol L−1, respectively. Furthermore, it is demonstrated that the activity pattern of MMPs in serum closely relevant to the disease progression and type of tumor.