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Published on: June 21, 2011
Investigating the specific uptake of EGF-conjugated nanoparticles in lung cancer cells using fluorescence imaging
Honglin Jin1, Jonathan F Lovell2, Juan Chen3
1Ontario Cancer Institute and Campbell Family Cancer Research Institute, University of Toronto, Toronto, Canada ; Department of Medical Biophysics, University of Toronto, Toronto, Canada ; Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, China.
Abstract:
Targeted nanoparticles have the potential to deliver a large drug payload specifically to cancer cells. Targeting requires that a ligand on the nanoparticle surface interact with a specific membrane receptor on target cells. However, the contribution of the targeting ligand to nanoparticle delivery is often influenced by non-specific nanoparticle uptake or secondary targeting mechanisms. In this study, we investigate the epidermal growth factor (EGF) receptor-targeting specificity of a nanoparticle by dual-color fluorescent labeling. The targeted nanoparticle was a fluorescently labeled, EGF-conjugated HDL-like peptide-phospholipid scaffold (HPPS) and the cell lines expressed EGF receptor linked with green fluorescent protein (EGFR-GFP). Using LDLA7 cells partially expressing EGFR-GFP, fluorescence imaging demonstrated the co-internalization of EGFR-GFP and EGF-HPPS, thus validating its targeting specificity. Furthermore, specific EGFR-mediated uptake of the EGF-HPPS nanoparticle was confirmed using human non-small cell lung cancer A549 cells. Subsequent confocal microscopy and flow cytometry studies delineated how secondary targeting mechanisms affected the EGFR targeting. Together, this study confirms the EGFR targeting of EGF-HPPS in lung cancer cells and provides insight on the potential influence of unintended targets on the desired ligand-receptor interaction.
Insights
This study validates the specific targeting of epidermal growth factor (EGF) receptor-positive lung cancer cells using EGF-conjugated nanoparticles (EGF-HPPS). It also explores how secondary mechanisms can impact targeted drug delivery efficacy.
Area of Science:
- Biomedical Engineering
- Nanotechnology
- Oncology
Background:
- Targeted nanoparticles offer potential for specific cancer drug delivery.
- Ligand-receptor interactions are crucial for nanoparticle targeting.
- Non-specific uptake and secondary mechanisms can affect targeting specificity.
Purpose of the Study:
- To investigate the epidermal growth factor (EGF) receptor-targeting specificity of EGF-conjugated HDL-like peptide-phospholipid scaffold (EGF-HPPS) nanoparticles.
- To confirm EGFR-mediated uptake in lung cancer cells.
- To understand the influence of secondary targeting mechanisms on EGFR targeting.
Main Methods:
- Dual-color fluorescent labeling of nanoparticles and cells expressing EGF receptor linked with green fluorescent protein (EGFR-GFP).
- Fluorescence imaging using LDLA7 cells partially expressing EGFR-GFP.
- Confirmation of uptake in human non-small cell lung cancer A549 cells.
- Confocal microscopy and flow cytometry to analyze secondary targeting effects.
Main Results:
- Fluorescence imaging confirmed co-internalization of EGFR-GFP and EGF-HPPS, validating targeting specificity.
- EGFR-mediated uptake of EGF-HPPS was confirmed in A549 lung cancer cells.
- Secondary targeting mechanisms were delineated, showing their potential influence on EGFR targeting.
Conclusions:
- The study confirms the EGFR targeting capability of EGF-HPPS nanoparticles in lung cancer cells.
- Insights were gained into how unintended targets may affect desired ligand-receptor interactions in targeted nanoparticle delivery.
