title: Molecular recognition of proteolytic activity in metastatic cancer cells using fluorogenic gold nanoprobes.
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.
authors.affiliation: Department of Biomedical Engineering, Yonsei University, Wonju, Gangwondo 220-710, Republic of Korea.
authors.forename: Yoochan
authors.initials: Y
authors.lastname: Hong
authors.affiliation: Department of Radiology, College of Medicine, Yonsei University, Seoul 120-752, Republic of Korea; Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul 120-752, Republic of Korea.
authors.forename: Minhee
authors.initials: M
authors.lastname: Ku
authors.affiliation: Department of Radiology, College of Medicine, Yonsei University, Seoul 120-752, Republic of Korea; Nanomedical National Core Research Center, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Dan
authors.initials: D
authors.lastname: Heo
authors.affiliation: Department of Radiology, College of Medicine, Yonsei University, Seoul 120-752, Republic of Korea; Nanomedical National Core Research Center, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Seungyeon
authors.initials: S
authors.lastname: Hwang
authors.affiliation: Department of Radiology, College of Medicine, Yonsei University, Seoul 120-752, Republic of Korea; Nanomedical National Core Research Center, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Eugene
authors.initials: E
authors.lastname: Lee
authors.affiliation: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.
authors.forename: Joseph
authors.initials: J
authors.lastname: Park
authors.affiliation: Department of Chemical and Biomolecular Engineering, Yonsei University, Seoul 120-749, Republic of Korea.
authors.forename: Jihye
authors.initials: J
authors.lastname: Choi
authors.affiliation: Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul 120-752, Republic of Korea; Institute of Endocrine Research, Department of Endocrinology, College of Medicine, Yonsei University, Seoul 120-752, Republic of Korea.
authors.forename: Hyeon Jung
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Severance Integrative Research Institute for Cerebral & Cardiovascular Diseases, Yonsei University Health System, Seoul 120-752, Republic of Korea.
authors.forename: Miran
authors.initials: M
authors.lastname: Seo
authors.affiliation: Brain Korea 21 PLUS Project for Medical Science, Yonsei University, Seoul 120-752, Republic of Korea; Institute of Endocrine Research, Department of Endocrinology, College of Medicine, Yonsei University, Seoul 120-752, Republic of Korea.
authors.forename: Eun Jig
authors.initials: EJ
authors.lastname: Lee
authors.affiliation: Department of Oral Pathology, Oral Cancer Research Institute, Yonsei University, Seoul 120-752, Republic of Korea.
authors.forename: Jong In
authors.initials: JI
authors.lastname: Yook
authors.affiliation: Department of Chemical and Biomolecular Engineering, Yonsei University, Seoul 120-749, Republic of Korea; YUHS-KRIBB Medical Convergence Research Institute, Seoul 120-752, Republic of Korea.
authors.forename: Seungjoo
authors.initials: S
authors.lastname: Haam
authors.affiliation: Department of Radiology, College of Medicine, Yonsei University, Seoul 120-752, Republic of Korea; YUHS-KRIBB Medical Convergence Research Institute, Seoul 120-752, Republic of Korea; Severance Biomedical Science Institute (SBSI), Seoul 120-752, Republic of Korea.
authors.forename: Yong-Min
authors.initials: YM
authors.lastname: Huh
authors.affiliation: Department of Biomedical Engineering, Yonsei University, Wonju, Gangwondo 220-710, Republic of Korea.
authors.forename: Dae Sung
authors.initials: DS
authors.lastname: Yoon
authors.affiliation: Department of Radiology, College of Medicine, Yonsei University, Seoul 120-752, Republic of Korea; YUHS-KRIBB Medical Convergence Research Institute, Seoul 120-752, Republic of Korea; Severance Biomedical Science Institute (SBSI), Seoul 120-752, Republic of Korea. Electronic address: jss@yuhs.ac.
authors.forename: Jin-Suck
authors.initials: JS
authors.lastname: Suh
authors.affiliation: Department of Radiology, College of Medicine, Yonsei University, Seoul 120-752, Republic of Korea; Severance Integrative Research Institute for Cerebral & Cardiovascular Diseases, Yonsei University Health System, Seoul 120-752, Republic of Korea; YUHS-KRIBB Medical Convergence Research Institute, Seoul 120-752, Republic of Korea. Electronic address: 177hum@yuhs.ac.
authors.forename: Jaemoon
authors.initials: J
authors.lastname: Yang
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D002465:Cell Movement; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D006801:Humans; D061986:MCF-7 Cells; D053511:Matrix Metalloproteinase 14 / Q000032:analysis* / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D009362:Neoplasm Metastasis / Q000175:diagnosis* / Q000473:pathology; D009369:Neoplasms / Q000175:diagnosis / Q000201:enzymology / Q000473:pathology*; D061848:Optical Imaging / Q000379:methods; D059748:Proteolysis
chemical_list: D005456:Fluorescent Dyes; D006046:Gold; C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14
keywords: Cancer metastasis; Fluorescence; Membrane type 1-matrix metalloproteinase (MT1-MMP); Nanoparticle surface energy transfer (NSET); Nanoprobe; Proteolysis