Endocytic pathway of vascular cell adhesion molecule 1 in human umbilical vein endothelial cell identified in vitro by using functionalized nontoxic fluorescent quantum dots

[Display omitted] •Custom designed VCAM1 binding peptides to target bind VCAM1.•Custom made 5-carboxyfluorescein (5FAM)-labeled VCAM1 binding peptides.•Nontoxic QDs in-house made/functionalized with VCAM1 binding peptides.•Fluorescence microscopy study of QDs and 5FAM in life cell in vitro.•VCAM1 in...

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Published in:	Sensors and actuators. B, Chemical Chemical, 2019, Vol.297, p.126702, Article 126702
Main Authors:	Fu, Ying, Jussi, Johnny, Wang, Qin, Brismar, Hjalmar, Liu, Yushen, Yang, Xifeng, Chen, Yun
Format:	Article
Language:	eng
Subjects:	Annan medicinteknik Atom and Molecular Physics and Optics Atom- och molekylfysik och optik Binding Biological and Biomedical Physics Biologisk och biomedicinsk fysik Bionanosensors Cell adhesion Cell adhesion & migration Cell and Molecular Biology Cell Biology Cell membranes Cell- och molekylärbiologi Cellbiologi Colloidal fluorescent quantum dot Deposition Endosome Endosomes Endothelial cells Fluorescence Human umbilical vein endothelial cell In vivo methods and tests Lysosome Lysosomes Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci) Other Medical Engineering Peptides Quantum dots Therapy Tumors Vascular cell adhesion molecule 1 (VCAM1)
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Summary:	[Display omitted] •Custom designed VCAM1 binding peptides to target bind VCAM1.•Custom made 5-carboxyfluorescein (5FAM)-labeled VCAM1 binding peptides.•Nontoxic QDs in-house made/functionalized with VCAM1 binding peptides.•Fluorescence microscopy study of QDs and 5FAM in life cell in vitro.•VCAM1 in HUVEC induced by TNFα is shown to be internalized into lysosomes.•Interdisciplinary research strategy & precise biosensing technique for life science Studies about vascular cell adhesion molecule 1 (VCAM1) in tumor growth, metastasis, and angiogenesis suggest that targeting VCAM1 expression is an attractive strategy for diagnosis and anti-tumor therapy. However, the endocytic pathway of VCAM1 in vascular cells has not been well characterized. In this study we visualize the endocytic pathway of tumor necrosis factor α (TNFα) induced VCAM1 in human umbilical vein endothelial cell (HUVEC) in vitro using 5-carboxyfluorescein labeled VCAM1 binding peptides and fluorescent water-dispersible 3-mercaptopropionic acid (3MPA)-coated CdSe-CdS/Cd0.5Zn0.5S/ZnS core–multishell nontoxic quantum dots (3MPA-QDs) functionalized with VCAM1 binding peptides. Clear key in vitro observations are as follows: (a) 3MPA-QDs functionalized with VCAM1 binding peptides, denoted as VQDs, adhered and aggregated cumulatively to cell membrane around 2 h after VQD deposition to cell culture medium and were found in lysosomes in TNFα-treated HUVECs approximately 24 h after VQD deposition; (b) VQDs remained in TNFα-treated HUVECs for the whole 16 days of the experimental observation period; (c) quite differently, 3MPA-QDs were endocytosed then exocytosed by HUVECs via endosomes in about 24–48 h after 3MPA-QD deposition. Our study suggests that VCAM1 molecules, initially expressed on cell membrane induced by TNFα treatment, are internalized into lysosomes. This provides a novel means to deliver materials to lysosomes such as enzyme replacement therapy. Moreover, our meticulous sensing methodology of devising fluorescent nontoxic QDs advances biosensing technique for studying cellular activities in vitro and in vivo.
ISSN:	0925-4005 1873-3077 1873-3077