A coumarin-based fluorescence resonance energy transfer probe targeting matrix metalloproteinase-2 for the detection of cervical cancer

Abstract

Cervical cancer is one of the most common gynecological malignancies worldwide. At present, the methods used for cervical cancer screening have many disadvantages. Matrix metalloproteinase-2 (MMP-2) is low or absent in normal cervical epithelial cells while overexpressed in cervical intraepithelial neoplasia and cervical cancer, which provides the possibility for the detection of cervical cancer based on MMP-2. The development of a strategy with high sensitivity and specificity for cervical cancer detection would be expected to improve the cure rate. Connecting a fluorophore [7-(diethylamino)-2-oxo-2H-chromene-3-succinimidyl ester] with a quencher {4-[4-(dimethylamino)phenylazo]benzoic acid N-succinimidyl ester} via an MMP-2 substrate peptide GPLGVRGKGG, a new coumarin-based fluorescence resonance energy transfer probe targeting MMP-2 was prepared to examine cervical cancer by cell imaging. The cervical cancer cell lines showed green fluorescence with intensities in the order of CaSki > SiHa > C33A > HeLa, which indicated that the cervical cancer cell lines possessed significantly differential expression levels of MMP-2. Quantitative (real-time) PCR, RT-PCR, and western blot experiments were consistent with such results. The fluorescence detection limit of the probe for MMP-2 was estimated to be 0.05 ng/ml. Therefore, this MMP-2 probe potentially provides a sensitive and specific visual method for cervical cancer screening, diagnosis and prognostic judgment.