Dual-Functional Liposomes with Carbonic Anhydrase IX Antibody and BR2 Peptide Modification Effectively Improve Intracellular Delivery of Cantharidin to Treat Orthotopic Hepatocellular Carcinoma Mice
Abstract
:1. Introduction
2. Results and Discussion
2.1. Preparation and Characterization of Dual-Functionalized Liposomes (DF-Lp)
2.2. Cell Association of NBD-DPPE-Labeled Dual-Functional Liposomes
2.3. Analysis of CA IX Antibody Directed Specific Cellular Uptake of Dual-Functional Liposomes
2.4. Analysis of CA IX Receptor-Mediated Endocytosis of Dual Functional Liposomes
2.5. Cellular Internalization of Dual Functional Liposomes by Cancer Cells
2.6. Orthotopic HCC Model for In Vivo Testing
2.7. In Vivo Tumor Distribution of DF-Lp
2.8. Tumor Growth Inhibition Study of DF-Lp/CTD
3. Materials and Methods
3.1. Materials
3.2. Cell Lines
3.3. Animals
3.4. Preparation of Single-Ligand and Dual-Ligand Modified Liposomes
3.4.1. Synthesis of DSPE-PEG-BR2 Conjugates
3.4.2. Synthesis of DSPE-PEG-CA IX Conjugated Micelle
3.4.3. Preparation and Characterization of Liposomal CTD
3.5. In Vitro Studies
3.5.1. Induction of CA IX Expression In Vitro
3.5.2. Cell Association of NBD-DPPE Labeled Dual-Functional Liposomes
3.5.3. Analysis of CA IX Antibody Directed Specific Cellular Uptake of Dual Functional Liposomes
3.5.4. Analysis of CA IX Receptor-Mediated Endocytosis of Dual-Functional Liposomes
3.5.5. Cellular Internalization of Dual-Functional Liposomes by Cancer Cells
3.6. In Vitro Cytotoxicity Study
3.7. In Vivo Performance of Therapeutic Efficacy of DF-Lp
3.7.1. Othotopic Tumor Model in Mice
3.7.2. Biodistribution Studies
3.7.3. Tumor Growth Inhibition Studies
3.8. Statistical Analysis
4. General Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Sample Availability: Not available. |
Liposome Type | Liposome Component (Molar Ratio) | Particle Size (nm) | PDI |
---|---|---|---|
CA IX-Lp/CTD | SPC/DSPE-PEG/DSPE-PEG-Mal (95.8:3.8:0.4) | 75.7 ± 1.5 | 0.135 ± 0.006 |
BR2-Lp/CTD | SPC/DSPE-PEG/DSPE-PEG-BR2 (96:2.0:2.0) | 92.4 ± 1.2 | 0.261 ± 0.022 |
DF-Lp/CTD | SPC/DSPE-PEG/DSPE-PEG-BR2/DSPE-PEG-Mal (95.2:1.9:2.0:0.87) | 98.3 ± 1.8 | 0.256 ± 0.003 |
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Zhang, X.; Lin, C.; Chan, W.; Liu, K.; Lu, A.; Lin, G.; Hu, R.; Shi, H.; Zhang, H.; Yang, Z. Dual-Functional Liposomes with Carbonic Anhydrase IX Antibody and BR2 Peptide Modification Effectively Improve Intracellular Delivery of Cantharidin to Treat Orthotopic Hepatocellular Carcinoma Mice. Molecules 2019, 24, 3332. https://doi.org/10.3390/molecules24183332
Zhang X, Lin C, Chan W, Liu K, Lu A, Lin G, Hu R, Shi H, Zhang H, Yang Z. Dual-Functional Liposomes with Carbonic Anhydrase IX Antibody and BR2 Peptide Modification Effectively Improve Intracellular Delivery of Cantharidin to Treat Orthotopic Hepatocellular Carcinoma Mice. Molecules. 2019; 24(18):3332. https://doi.org/10.3390/molecules24183332
Chicago/Turabian StyleZhang, Xue, Congcong Lin, Waikei Chan, Kanglun Liu, Aiping Lu, Ge Lin, Rong Hu, Hongcan Shi, Hongqi Zhang, and Zhijun Yang. 2019. "Dual-Functional Liposomes with Carbonic Anhydrase IX Antibody and BR2 Peptide Modification Effectively Improve Intracellular Delivery of Cantharidin to Treat Orthotopic Hepatocellular Carcinoma Mice" Molecules 24, no. 18: 3332. https://doi.org/10.3390/molecules24183332